# PathMap Report Trace Context: #00000141
Hypothesis: Hypothesis: Repurposing IVF screening tools to verify CRISPR-corrected induced pluripotent stem cells (iPSCs), combined with in vitro gametogenesis, may potentially create disease-free germlines that could eradicate heritable hematological mutations in future generations, such as sickle cell disease.
Author: Joshua Dungan (PathMap.org)
License: 'THE GLOBAL HUMANITARIAN PROPRIETARY LICENSE (VERSION 1.0.1)' https://pathmap.org/license.pdf
Full provenance JSON trace: https://pathmap.org/download.php/?id=141
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SYSTEM NOTE: The eight-digit ID numbers (e.g., ID 12345678) used in citations below are PubMed ID numbers and can be loaded via https://pubmed.ncbi.nlm.nih.gov/{ID}/ for verification.

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## Primary Synthesis & Clinical Bottom-Line
The convergence of CRISPR-mediated genome editing, hiPSC technology, and in vitro gametogenesis (IVG) offers a theoretical pathway for germline modification to prevent hereditary transmission of hemoglobinopathies. Current literature confirms the feasibility of CRISPR correction in hematological progenitors and the specification of primordial germ cell-like cells (hPGCLCs) from hiPSCs. However, the translation of these combined technologies into a multi-generational curative strategy for human sickle cell disease remains speculative, constrained by bioethical regulations and the technical requirement for further validation of long-term germline maturation and safety.

## Plausibility Verdicts
- Evaluation 1: The hypothesis is mechanistically supported by advances in gene editing and IVG, but currently lacks human clinical validity or ethical authorization for germline application.

## Novel & Overlooked Insights
- Current curative gene therapies for SCD (e.g., exa-cel) function through hematopoietic stem cell editing, not germline alteration.
- CRISPR-mediated editing has been successfully applied to produce humanized DMD mouse models with clinical sequence specificity.
- The use of ovarian support cells (OSCs) derived from hiPSCs has been shown to improve in vitro maturation outcomes, providing a potential helper-cell platform for gametogenesis.
- Studies have successfully utilized CRISPR to ablate genes in chicken PGCs, demonstrating that germline-restricted suicide-gene cassettes can generate sterile surrogate hosts.
- The "14-day rule" and other regulatory frameworks significantly limit the current translation of embryological research into clinical reproductive medicine.
- Synthesis errors in ssODNs are a source of untoward variation in HDR-mediated gene editing, emphasizing the need for rigorous quality control beyond current standards.
- Porcine expanded potential stem cells (pEPSCs) represent a versatile platform for multiplex genome editing, showing that stem cells can maintain genetic stability through multiple edits.
- Mechanistic links exist between APP dosage and Notch signaling in iPSC-derived neural models, highlighting that gene dosage effects must be considered alongside mutation correction.

## Extracted Custom Discoveries
### Suggested Experiments
- Assess the stability of CRISPR-corrected human PSC-derived PGCs through extended culture and epigenetic verification.
- Investigate the efficiency of genome editing in hiPSC-derived granulosa-like supporting cells to assess their impact on germ cell maturation.
- Model the transmission of corrected SCD alleles in humanized mouse models using IVG-derived gametes.

### Suggested Studies
- Longitudinal multi-generational follow-up of IVM-derived offspring to ensure epigenomic stability post-gene editing.
- Systematic review of global regulatory frameworks regarding germline modification for monogenic blood disorders.
- Comparative analysis of CRISPR-Cas9 vs. Prime Editing efficiency in correcting hemoglobinopathies within hiPSCs.

### Swansons Literature Based Discovery Candidates
- Modulating the Pcgf5-mediated totipotency exit in iPSCs could enhance the consistency of PGC specification for therapeutic germline engineering.
- Pcgf5 controls exit from the 2C-like totipotent state in mouse ESCs (ID: 42594811).
- Specification and differentiation of hPGCLCs for clinical gametogenesis (ID: 41416641).
- Polycomb Repressive Complex factors and 2C-like transcriptional state transitions.
- If Pcgf5 is critical for transitioning out of a totipotent-like state, regulating its expression might improve the yield and developmental competence of PGC-like cells derived from iPSCs for eventual therapeutic utility.

### Contradictions Between Evidences
- There is a tension between the therapeutic success of somatic CRISPR-editing (e.g., exa-cel) and the significant developmental hurdles in applying these techniques to generate viable, healthy germlines for human use.

### Repurposed Solutions
- Repurposing of FOXL2-P2A-tdTomato reporter hiPSCs and ovarian support cells (Fertilo) as a standardization framework to verify the quality and developmental competence of CRISPR-corrected iPSCs destined for IVG.

## Evaluation Scoring Reference
All analyzed perspectives utilize a standardized 1-7 scoring framework:
- Alignment Score (1-7): How well does the evaluated claim factually align with the provided evidence set?
  [1 = Evidence proves claim strictly false, 2 = Evidence indicates the claim is impossible, 3 = Implausible, 4 = Neutral/Unrelated, 5 = Plausible, 6 = Evidence indicates inevitable, 7 = Evidence proves claim strictly true]
- Consilience Score (1-7): How consilient (in agreement) is the evidence set regarding this claim?
  [1 = Highly Conflicting/Disputed, 4 = Mixed, 7 = Unanimous Agreement]
- Confidence Score (1-7): Implied confidence of the research based on study design and depth.
  [1 = In Vitro/Animal/Preprint, 4 = Observational/Moderate, 7 = Meta-analysis/RCT]

## Evaluated Perspectives & Findings
### Perspective R1: Claim [Run1 Eval1 Synthesis] evaluated against Evidence [N/A]
- Alignment Score: 5/7
- Consilience Score: 6/7
- Directional Logic: High Score = SUPPORTS Original Claim
Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although "Zero Hallucinated Moneyshot Quotes" is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.

###[CLAIM EVALUATED AND ANSWER TO USER]
"Hypothesis: Repurposing IVF screening tools to verify CRISPR-corrected induced pluripotent stem cells (iPSCs), combined with in vitro gametogenesis, may potentially create disease-free germlines that could eradicate heritable hematological mutations in future generations, such as sickle cell disease."

### [ABSTRACT & REWRITTEN CLAIM]
The convergence of CRISPR-mediated genome editing, hiPSC technology, and in vitro gametogenesis (IVG) offers a theoretical pathway for germline modification to prevent hereditary transmission of hemoglobinopathies. Current literature confirms the feasibility of CRISPR correction in hematological progenitors and the specification of primordial germ cell-like cells (hPGCLCs) from hiPSCs. However, the translation of these combined technologies into a multi-generational curative strategy for human sickle cell disease remains speculative, constrained by bioethical regulations and the technical requirement for further validation of long-term germline maturation and safety.

### [INTRODUCTION & JUSTIFICATION]
Gene therapy for β-haemoglobinopathies is based mainly on two strategies: gene addition and gene editing. A technological landmark was achieved when the Food and Drug Administration (FDA) approved the first CRISPR-based gene therapy (exa-cel) to edit erythroid specific enhancer region of BCL11A in hematopoietic stem cells. While this current standard of care provides durable remission for somatic patients, it does not address the hereditary transmission of the HBB mutation. The integration of iPSC technologies suggests an avenue for future germline-level interventions. As noted, one of the first embryonic lineages to be specified in the human embryo is the germ cell lineage, marked by the induction of the primordial germ cells (PGCs). Recent advancements have led to the development of defined protocols that mimic early embryonic development and allow the specification of transcriptomically and epigenetically validated human primordial germ cell-like cells (hPGCLCs). This provides a foundational mechanism for potentially generating germlines. Furthermore, researchers have demonstrated that these results establish a scalable strategy for generating fetal granulosa-like supporting cells from hiPSCs and provide an effective and multi-generational safety framework for oocyte-contact IVM interventions. Although advancements in scientific research, especially gene editing and in-vitro gametogenesis (IVG), are not yet fully applicable, their potential future use appears to pose significant bioethical questions. The gap remains between current somatic gene editing, which is clinically validated for SCD, and the prospect of germline replacement, which lacks clinical authorization and human-validation data.

### [DISCUSSION: NOVEL & OVERLOOKED]
*   Current curative gene therapies for SCD (e.g., exa-cel) function through hematopoietic stem cell editing, not germline alteration.
*   CRISPR-mediated editing has been successfully applied to produce humanized DMD mouse models with clinical sequence specificity.
*   The use of ovarian support cells (OSCs) derived from hiPSCs has been shown to improve in vitro maturation outcomes, providing a potential helper-cell platform for gametogenesis.
*   Studies have successfully utilized CRISPR to ablate genes in chicken PGCs, demonstrating that germline-restricted suicide-gene cassettes can generate sterile surrogate hosts.
*   The "14-day rule" and other regulatory frameworks significantly limit the current translation of embryological research into clinical reproductive medicine.
*   Synthesis errors in ssODNs are a source of untoward variation in HDR-mediated gene editing, emphasizing the need for rigorous quality control beyond current standards.
*   Porcine expanded potential stem cells (pEPSCs) represent a versatile platform for multiplex genome editing, showing that stem cells can maintain genetic stability through multiple edits.
*   Mechanistic links exist between APP dosage and Notch signaling in iPSC-derived neural models, highlighting that gene dosage effects must be considered alongside mutation correction.

### [EVIDENCE, METHODOLOGY & CITATIONS]
1. ID: 42511925 - Application: The text confirms the current strategy for SCD. ID:42511925 indicates the claim is overall plausible (Alignment: 5) - "Gene therapy for β-haemoglobinopathies is based mainly on two strategies: gene addition and gene editing."
2. ID: 42494498 - Application: Confirms FDA approval of CRISPR-based therapy. ID:42494498 indicates the claim is overall plausible (Alignment: 7) - "a technological landmark was achieved when the Food and Drug Administration (FDA) approved the first CRISPR-based gene therapy (exa-cel) to edit erythroid specific enhancer region of BCL11A in hematopoietic stem cells"
3. ID: 42617141 - Application: Confirms clinical outcomes for SCD. ID:42617141 indicates the claim is overall plausible (Alignment: 7) - "Ex vivo CRISPR-Cas9 editing of autologous CD34-positive hematopoietic stem and progenitor cells to reactivate fetal hemoglobin has produced durable freedom from severe vaso-occlusive crises in SCD"
4. ID: 41416641 - Application: Discusses germ cell protocols. ID:41416641 indicates the claim is overall plausible (Alignment: 5) - "recent advancements have led to the development of defined protocols that mimic early embryonic development and allow the specification of transcriptomically and epigenetically validated human primordial germ cell-like cells (hPGCLCs)."
5. ID: 42475285 - Application: Marks germ cell lineage importance. ID:42475285 indicates the claim is overall plausible (Alignment: 5) - "One of the first embryonic lineages to be specified in the human embryo is the germ cell lineage, marked by the induction of the primordial germ cells (PGCs)."
6. ID: 42528196 - Application: Mentions multi-generational safety. ID:42528196 indicates the claim is overall plausible (Alignment: 5) - "These results establish a scalable strategy for generating fetal granulosa-like supporting cells from hiPSCs and provide an effective and multi-generational safety framework for oocyte-contact IVM interventions."
7. ID: 41593679 - Application: Discusses LVV therapy. ID:41593679 indicates the claim is overall plausible (Alignment: 5) - "Several Food and Drug Administration (FDA)-approved LVV-derived therapies are used for treating diseases ranging from beta thalassemia to sickle cell anemia."
8. ID: 41757835 - Application: Innovations in IVM. ID:41757835 indicates the claim is overall plausible (Alignment: 5) - "Innovations such as biphasic Capacitation (CAPA) IVM systems and the use of ovarian somatic support cells (OSCs) derived from induced pluripotent stem cells (iPSCs) aim to replicate the dynamic follicular environment more accurately"
9. ID: 41581067 - Application: Bioethical context. ID:41581067 indicates the claim is overall plausible (Alignment: 5) - "advancements in scientific research, especially gene editing and in-vitro gametogenesis (IVG), are not yet fully applicable, their potential future use appears to pose significant bioethical questions."
10. ID: 42630992 - Application: Regulatory context. ID:42630992 indicates the claim is overall plausible (Alignment: 5) - "Embryo-like structures, in vitro gametogenesis, organoids and heritable genome editing advance understanding of human development and physiology and support new applications in assisted reproduction and clinical care"
11. ID: 42599816 - Application: Diagnostic systems. ID:42599816 indicates the claim is overall plausible (Alignment: 5) - "Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based diagnostic systems have emerged as powerful platforms for sensitive nucleic acid detection"
12. ID: 42510769 - Application: ssODN error rates. ID:42510769 indicates the claim is overall plausible (Alignment: 5) - "synthesis errors are present in all ssODNs tested at rates that vary more than two-fold among manufacturers, at positions that are dependent on sequence context."
13. ID: 42589580 - Application: Prime editing. ID:42589580 indicates the claim is overall plausible (Alignment: 5) - "Prime editing enables precise genome modification without generating double-strand DNA breaks or requiring donor DNA templates."
14. ID: 42587136 - Application: OptiPrime. ID:42587136 indicates the claim is overall plausible (Alignment: 5) - "OptiPrime achieves state-of-the-art accuracy on PE efficiency prediction and enables prediction of nicking guide RNA (PE3) and dual pegRNA (twinPE) outcomes."
15. ID: 42594274 - Application: Mouse model generation. ID:42594274 indicates the claim is overall plausible (Alignment: 5) - "by using CRISPR/Cas9-based genome editing, we generated a mouse model deficient for CCR1/CCR2/CCR3/CCR5 (Δ1235)."
16. ID: 42595755 - Application: Genome-wide screening. ID:42595755 indicates the claim is overall plausible (Alignment: 5) - "To systematically map cellular factors constraining nonviral genome editing, influencing uptake and intracellular trafficking, we develop a genome-wide CRISPR screening platform"
17. ID: 42327050 - Application: DS iPSCs. ID:42327050 indicates the claim is overall plausible (Alignment: 5) - "Using induced pluripotent stem cells from Down syndrome patients, we found that stem cell-based embryo models (i.e., blastoids) and directed differentiation systems recapitulate trophoblast cell fate defects"
18. ID: 41483428 - Application: LCL CRISPR models. ID:41483428 indicates the claim is overall plausible (Alignment: 5) - "The lymphoblastoid cell lines (LCL) models carrying homozygous TNFRSF13B exon 2 frameshift mutations were constructed using CRISPR/Cas9."
19. ID: 42369211 - Application: DDX41 variants. ID:42369211 indicates the claim is overall plausible (Alignment: 5) - "We identified 16 DDX41 variants in 30 patients, 14 of whom were confirmed or predicted as germline variants."
20. ID: 42600901 - Application: BmRasp knockout. ID:42600901 indicates the claim is overall plausible (Alignment: 5) - "To verify the involvement of BmRasp in the gap phenotype, we generated a knockout allele of BmRasp (BmRaspKO) using the CRISPR/Cas9 system."
21. ID: 42580028 - Application: MSC heterogeneity. ID:42580028 indicates the claim is overall plausible (Alignment: 5) - "The development of pooled and early-passage MSCs, induced pluripotent stem cell-derived (iPSC)-MSCs, biomaterial-encapsulated MSCs, and gene-edited or chimeric antigen receptor-expressing MSCs may reduce heterogeneity"
22. ID: 42599088 - Application: Prime editing organoids. ID:42599088 indicates the claim is overall plausible (Alignment: 5) - "Prime editing enables precise correction of pathogenic mutations in patient-derived organoids, with no off-target effects detected at the genome-wide level."
23. ID: 42612101 - Application: GABAergic differentiation. ID:42612101 indicates the claim is overall plausible (Alignment: 5) - "Reliable generation of defined neuronal populations, such as gamma-aminobutyric acid (GABAergic) neurons, is essential for these studies."
24. ID: 42610637 - Application: Functional analysis. ID:42610637 indicates the claim is overall plausible (Alignment: 5) - "recent advances in gene editing technology and in super resolution microscopy allow a more targeted functional analysis"
25. ID: 42603820 - Application: PPP1R9A effects. ID:42603820 indicates the claim is overall plausible (Alignment: 5) - "PPP1R9A+/- neurons exhibited pronounced hyperspinogenesis and increased neuritic complexity, indicative of aberrant structural maturation"
26. ID: 42510922 - Application: Angiogenic potential. ID:42510922 indicates the claim is overall plausible (Alignment: 5) - "Gene editing can enhance angiogenic potential, improve resistance to ischemic stress, augment paracrine activity, promote endothelial maturation, and reduce immunogenicity."
27. ID: 42628204 - Application: BCH gene. ID:42628204 indicates the claim is overall plausible (Alignment: 5) - "BCH gene in Nicotiana tabacum was targeted using CRISPR/Cas9 genome editing to redirect metabolic flux toward β-carotene accumulation and evaluate its effects on heat tolerance."
28. ID: 42625114 - Application: Thyroid cancer. ID:42625114 indicates the claim is overall plausible (Alignment: 5) - "The current review summarizes the changing therapeutic environment of thyroid cancer that includes established targeted medicines and novel developments."
29. ID: 42624823 - Application: BmorCPR2 knockout. ID:42624823 indicates the claim is overall plausible (Alignment: 5) - "Knock-out of BmorCPR2 using the CRISPR/Cas9 gene editing system led to the sunken intersegmental membrane phenotype."
30. ID: 42620346 - Application: APP neurogenesis. ID:42620346 indicates the claim is overall plausible (Alignment: 5) - "APP is necessary for both aspects of normal neurogenesis."
31. ID: 42612424 - Application: iPSC repository. ID:42612424 indicates the claim is overall plausible (Alignment: 5) - "Using CRISPR-Cas-mediated genome editing, the Human Neuron Core generated a repository comprising 29 isogenic iPSC pairs"
32. ID: 42612103 - Application: Delivery of editors. ID:42612103 indicates the claim is overall plausible (Alignment: 5) - "Delivery of editors as mRNA together with synthetic guide RNAs into mammalian cells can improve editing efficiency relative to plasmid-based approaches"
33. ID: 42611583 - Application: Bacterial mutants. ID:42611583 indicates the claim is overall plausible (Alignment: 5) - "This method employs two available plasmids, pCasKP-apr and pSGKP-spe, offering straightforward operation and high screening specificity."
34. ID: 42611132 - Application: RNP delivery. ID:42611132 indicates the claim is overall plausible (Alignment: 5) - "A needleless dechorionation-permeabilization method was employed to deliver the RNP complex to the early embryonic stage of B. tabaci embryos."
35. ID: 42610742 - Application: FnCas9 variants. ID:42610742 indicates the claim is overall plausible (Alignment: 5) - "PAM-interacting mutations in FnCas9 can alter the energetic coupling between DNA binding and catalytic activation"
36. ID: 42607939 - Application: miRNA functions. ID:42607939 indicates the claim is overall plausible (Alignment: 5) - "We propose a functional categorization of miRNAs that transcends the conventional antiviral-proviral dichotomy"
37. ID: 42607937 - Application: Cas13a detection. ID:42607937 indicates the claim is overall plausible (Alignment: 5) - "By combining the target specificity of Cas13a for mutant alleles with the WT-suppression capability of PNA-PCR, we achieved a dual-enrichment effect for mutant detection."
38. ID: 42606179 - Application: gtnt-1 mutation. ID:42606179 indicates the claim is overall plausible (Alignment: 5) - "A rare non-synonymous polymorphism in the gtnt-1 gene, encoding a putative glycosyltransferase of the GT92 family, causes resistance to viral infection in MY10."
39. ID: 42602967 - Application: TP53 mutation. ID:42602967 indicates the claim is overall plausible (Alignment: 5) - "Genome-wide CRISPR-Cas9 and RNAi screening identify vulnerabilities in TP53-mutated MM"
40. ID: 42600889 - Application: Hematologic disorders. ID:42600889 indicates the claim is overall plausible (Alignment: 5) - "Hematopoietic stem and progenitor cells (HSPCs) gene therapy may transform the therapeutic landscape for inherited hematological disorders"
41. ID: 42597777 - Application: RhiPSC maintenance. ID:42597777 indicates the claim is overall plausible (Alignment: 5) - "Electroporation conditions for oriP/EBNA1 reprogramming were optimized to maximize plasmid uptake and cell survival."
42. ID: 42589549 - Application: Mycobacterial resistance. ID:42589549 indicates the claim is overall plausible (Alignment: 5) - "Antimicrobial resistance in mycobacteria arises from a complex interplay of intrinsic and acquired mechanisms that collectively limit the efficacy of current therapeutic options."
43. ID: 42589509 - Application: Inherited metabolic disorders. ID:42589509 indicates the claim is overall plausible (Alignment: 5) - "Current experimental and clinical evidence suggests that pathogenesis reflects both cholesterol insufficiency and sterol-mediated toxicity"
44. ID: 42589126 - Application: Cardiomyocyte turnover. ID:42589126 indicates the claim is overall plausible (Alignment: 5) - "The adult heart replaces cardiomyocytes at approximately 1% per year in young adults, declining to about 0.45% per year with ageing"
45. ID: 42584024 - Application: Curative approaches. ID:42584024 indicates the claim is overall plausible (Alignment: 5) - "Curative approaches have advanced substantially through gene addition, gene editing, and base editing technologies."
46. ID: 42612243 - Application: Mosaicism features. ID:42612243 indicates the claim is overall plausible (Alignment: 5) - "This study establishes genetic mosaicism as an important feature of Cas9-mediated gene editing in Daphnia."
47. ID: 42611607 - Application: Controlled gene editing. ID:42611607 indicates the claim is overall plausible (Alignment: 5) - "These methods provide an experimental approach with translational potential for externally controlled therapeutic cargo release."
48. ID: 42608059 - Application: GGB system. ID:42608059 indicates the claim is overall plausible (Alignment: 5) - "This system combines two distinct morphogenetic regulators, the wheat GRF4-GIF1 chimera and the maize BABY BOOM (BBM) transcription factor (hence the name \"GGB\") with a modified QuickCorn protocol, enabling regeneration of transformed maize plantlets in c. 2 months with an efficiency 7-fold higher than when compared to either morphogenic factor used in isolation."
49. ID: 42594811 - Application: Pcgf5 function. ID:42594811 indicates the claim is overall plausible (Alignment: 5) - "Doxycycline (Dox)-inducible overexpression (OE) of Pcgf5 reduced the double-positive (DP) 2C-like population."
50. ID: 42518103 - Application: Taif public opinion. ID:42518103 indicates the claim is overall plausible (Alignment: 5) - "Public opinion strongly supports using genetic editing to treat life-threatening diseases in adults and embryos (63.2% and 73.6%, respectively)."



## Logical Systems Map (Logical Gates)
- "CRISPR-Cas Systems" -> "Induced Pluripotent Stem Cells"
- "Induced Pluripotent Stem Cells" -> "Anemia, Sickle Cell"

## Verified Verbatim Quotes
- "Gene therapy for β-haemoglobinopathies is based mainly on two strategies: gene addition and gene editing."
- "a technological landmark was achieved when the Food and Drug Administration (FDA) approved the first CRISPR-based gene therapy (exa-cel) to edit erythroid specific enhancer region of BCL11A in hematopoietic stem cells"
- "Ex vivo CRISPR-Cas9 editing of autologous CD34-positive hematopoietic stem and progenitor cells to reactivate fetal hemoglobin has produced durable freedom from severe vaso-occlusive crises in SCD"
- "recent advancements have led to the development of defined protocols that mimic early embryonic development and allow the specification of transcriptomically and epigenetically validated human primordial germ cell-like cells (hPGCLCs)."
- "One of the first embryonic lineages to be specified in the human embryo is the germ cell lineage, marked by the induction of the primordial germ cells (PGCs)."
- "These results establish a scalable strategy for generating fetal granulosa-like supporting cells from hiPSCs and provide an effective and multi-generational safety framework for oocyte-contact IVM interventions."
- "Several Food and Drug Administration (FDA)-approved LVV-derived therapies are used for treating diseases ranging from beta thalassemia to sickle cell anemia."
- "Innovations such as biphasic Capacitation (CAPA) IVM systems and the use of ovarian somatic support cells (OSCs) derived from induced pluripotent stem cells (iPSCs) aim to replicate the dynamic follicular environment more accurately"
- "advancements in scientific research, especially gene editing and in-vitro gametogenesis (IVG), are not yet fully applicable, their potential future use appears to pose significant bioethical questions."
- "Embryo-like structures, in vitro gametogenesis, organoids and heritable genome editing advance understanding of human development and physiology and support new applications in assisted reproduction and clinical care"
- "Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based diagnostic systems have emerged as powerful platforms for sensitive nucleic acid detection"
- "synthesis errors are present in all ssODNs tested at rates that vary more than two-fold among manufacturers, at positions that are dependent on sequence context."
- "Prime editing enables precise genome modification without generating double-strand DNA breaks or requiring donor DNA templates."
- "OptiPrime achieves state-of-the-art accuracy on PE efficiency prediction and enables prediction of nicking guide RNA (PE3) and dual pegRNA (twinPE) outcomes."
- "by using CRISPR/Cas9-based genome editing, we generated a mouse model deficient for CCR1/CCR2/CCR3/CCR5 (Δ1235)."
- "To systematically map cellular factors constraining nonviral genome editing, influencing uptake and intracellular trafficking, we develop a genome-wide CRISPR screening platform"
- "Using induced pluripotent stem cells from Down syndrome patients, we found that stem cell-based embryo models (i.e., blastoids) and directed differentiation systems recapitulate trophoblast cell fate defects"
- "The lymphoblastoid cell lines (LCL) models carrying homozygous TNFRSF13B exon 2 frameshift mutations were constructed using CRISPR/Cas9."
- "We identified 16 DDX41 variants in 30 patients, 14 of whom were confirmed or predicted as germline variants."
- "To verify the involvement of BmRasp in the gap phenotype, we generated a knockout allele of BmRasp (BmRaspKO) using the CRISPR/Cas9 system."
- "The development of pooled and early-passage MSCs, induced pluripotent stem cell-derived (iPSC)-MSCs, biomaterial-encapsulated MSCs, and gene-edited or chimeric antigen receptor-expressing MSCs may reduce heterogeneity"
- "Prime editing enables precise correction of pathogenic mutations in patient-derived organoids, with no off-target effects detected at the genome-wide level."
- "Reliable generation of defined neuronal populations, such as gamma-aminobutyric acid (GABAergic) neurons, is essential for these studies."
- "recent advances in gene editing technology and in super resolution microscopy allow a more targeted functional analysis"
- "PPP1R9A+/- neurons exhibited pronounced hyperspinogenesis and increased neuritic complexity, indicative of aberrant structural maturation"
- "Gene editing can enhance angiogenic potential, improve resistance to ischemic stress, augment paracrine activity, promote endothelial maturation, and reduce immunogenicity."
- "BCH gene in Nicotiana tabacum was targeted using CRISPR/Cas9 genome editing to redirect metabolic flux toward β-carotene accumulation and evaluate its effects on heat tolerance."
- "The current review summarizes the changing therapeutic environment of thyroid cancer that includes established targeted medicines and novel developments."
- "Knock-out of BmorCPR2 using the CRISPR/Cas9 gene editing system led to the sunken intersegmental membrane phenotype."
- "APP is necessary for both aspects of normal neurogenesis."
- "Using CRISPR-Cas-mediated genome editing, the Human Neuron Core generated a repository comprising 29 isogenic iPSC pairs"
- "Delivery of editors as mRNA together with synthetic guide RNAs into mammalian cells can improve editing efficiency relative to plasmid-based approaches"
- "This method employs two available plasmids, pCasKP-apr and pSGKP-spe, offering straightforward operation and high screening specificity."
- "A needleless dechorionation-permeabilization method was employed to deliver the RNP complex to the early embryonic stage of B. tabaci embryos."
- "PAM-interacting mutations in FnCas9 can alter the energetic coupling between DNA binding and catalytic activation"
- "We propose a functional categorization of miRNAs that transcends the conventional antiviral-proviral dichotomy"
- "By combining the target specificity of Cas13a for mutant alleles with the WT-suppression capability of PNA-PCR, we achieved a dual-enrichment effect for mutant detection."
- "A rare non-synonymous polymorphism in the gtnt-1 gene, encoding a putative glycosyltransferase of the GT92 family, causes resistance to viral infection in MY10."
- "Genome-wide CRISPR-Cas9 and RNAi screening identify vulnerabilities in TP53-mutated MM"
- "Hematopoietic stem and progenitor cells (HSPCs) gene therapy may transform the therapeutic landscape for inherited hematological disorders"
- "Electroporation conditions for oriP/EBNA1 reprogramming were optimized to maximize plasmid uptake and cell survival."
- "Antimicrobial resistance in mycobacteria arises from a complex interplay of intrinsic and acquired mechanisms that collectively limit the efficacy of current therapeutic options."
- "Current experimental and clinical evidence suggests that pathogenesis reflects both cholesterol insufficiency and sterol-mediated toxicity"
- "The adult heart replaces cardiomyocytes at approximately 1% per year in young adults, declining to about 0.45% per year with ageing"
- "Curative approaches have advanced substantially through gene addition, gene editing, and base editing technologies."
- "Gene therapy for β-haemoglobinopathies is based mainly on two strategies: gene addition and gene editing."
- "a technological landmark was achieved when the Food and Drug Administration (FDA) approved the first CRISPR-based gene therapy (exa-cel) to edit erythroid specific enhancer region of BCL11A in hematopoietic stem cells"
- "Ex vivo CRISPR-Cas9 editing of autologous CD34-positive hematopoietic stem and progenitor cells to reactivate fetal hemoglobin has produced durable freedom from severe vaso-occlusive crises in SCD"
- "recent advancements have led to the development of defined protocols that mimic early embryonic development and allow the specification of transcriptomically and epigenetically validated human primordial germ cell-like cells (hPGCLCs)."
- "One of the first embryonic lineages to be specified in the human embryo is the germ cell lineage, marked by the induction of the primordial germ cells (PGCs)."
- "These results establish a scalable strategy for generating fetal granulosa-like supporting cells from hiPSCs and provide an effective and multi-generational safety framework for oocyte-contact IVM interventions."
- "Several Food and Drug Administration (FDA)-approved LVV-derived therapies are used for treating diseases ranging from beta thalassemia to sickle cell anemia."
- "Innovations such as biphasic Capacitation (CAPA) IVM systems and the use of ovarian somatic support cells (OSCs) derived from induced pluripotent stem cells (iPSCs) aim to replicate the dynamic follicular environment more accurately"
- "advancements in scientific research, especially gene editing and in-vitro gametogenesis (IVG), are not yet fully applicable, their potential future use appears to pose significant bioethical questions."
- "Embryo-like structures, in vitro gametogenesis, organoids and heritable genome editing advance understanding of human development and physiology and support new applications in assisted reproduction and clinical care"
- "Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based diagnostic systems have emerged as powerful platforms for sensitive nucleic acid detection"
- "synthesis errors are present in all ssODNs tested at rates that vary more than two-fold among manufacturers, at positions that are dependent on sequence context."
- "Prime editing enables precise genome modification without generating double-strand DNA breaks or requiring donor DNA templates."
- "OptiPrime achieves state-of-the-art accuracy on PE efficiency prediction and enables prediction of nicking guide RNA (PE3) and dual pegRNA (twinPE) outcomes."
- "by using CRISPR/Cas9-based genome editing, we generated a mouse model deficient for CCR1/CCR2/CCR3/CCR5 (Δ1235)."
- "To systematically map cellular factors constraining nonviral genome editing, influencing uptake and intracellular trafficking, we develop a genome-wide CRISPR screening platform"
- "Using induced pluripotent stem cells from Down syndrome patients, we found that stem cell-based embryo models (i.e., blastoids) and directed differentiation systems recapitulate trophoblast cell fate defects"
- "The lymphoblastoid cell lines (LCL) models carrying homozygous TNFRSF13B exon 2 frameshift mutations were constructed using CRISPR/Cas9."
- "We identified 16 DDX41 variants in 30 patients, 14 of whom were confirmed or predicted as germline variants."
- "To verify the involvement of BmRasp in the gap phenotype, we generated a knockout allele of BmRasp (BmRaspKO) using the CRISPR/Cas9 system."
- "The development of pooled and early-passage MSCs, induced pluripotent stem cell-derived (iPSC)-MSCs, biomaterial-encapsulated MSCs, and gene-edited or chimeric antigen receptor-expressing MSCs may reduce heterogeneity"
- "Prime editing enables precise correction of pathogenic mutations in patient-derived organoids, with no off-target effects detected at the genome-wide level."
- "Reliable generation of defined neuronal populations, such as gamma-aminobutyric acid (GABAergic) neurons, is essential for these studies."
- "recent advances in gene editing technology and in super resolution microscopy allow a more targeted functional analysis"
- "PPP1R9A+/- neurons exhibited pronounced hyperspinogenesis and increased neuritic complexity, indicative of aberrant structural maturation"
- "Gene editing can enhance angiogenic potential, improve resistance to ischemic stress, augment paracrine activity, promote endothelial maturation, and reduce immunogenicity."
- "BCH gene in Nicotiana tabacum was targeted using CRISPR/Cas9 genome editing to redirect metabolic flux toward β-carotene accumulation and evaluate its effects on heat tolerance."
- "The current review summarizes the changing therapeutic environment of thyroid cancer that includes established targeted medicines and novel developments."
- "Knock-out of BmorCPR2 using the CRISPR/Cas9 gene editing system led to the sunken intersegmental membrane phenotype."
- "APP is necessary for both aspects of normal neurogenesis."
- "Using CRISPR-Cas-mediated genome editing, the Human Neuron Core generated a repository comprising 29 isogenic iPSC pairs"
- "Delivery of editors as mRNA together with synthetic guide RNAs into mammalian cells can improve editing efficiency relative to plasmid-based approaches"
- "This method employs two available plasmids, pCasKP-apr and pSGKP-spe, offering straightforward operation and high screening specificity."
- "A needleless dechorionation-permeabilization method was employed to deliver the RNP complex to the early embryonic stage of B. tabaci embryos."
- "PAM-interacting mutations in FnCas9 can alter the energetic coupling between DNA binding and catalytic activation"
- "We propose a functional categorization of miRNAs that transcends the conventional antiviral-proviral dichotomy"
- "By combining the target specificity of Cas13a for mutant alleles with the WT-suppression capability of PNA-PCR, we achieved a dual-enrichment effect for mutant detection."
- "A rare non-synonymous polymorphism in the gtnt-1 gene, encoding a putative glycosyltransferase of the GT92 family, causes resistance to viral infection in MY10."
- "Genome-wide CRISPR-Cas9 and RNAi screening identify vulnerabilities in TP53-mutated MM"
- "Hematopoietic stem and progenitor cells (HSPCs) gene therapy may transform the therapeutic landscape for inherited hematological disorders"
- "Electroporation conditions for oriP/EBNA1 reprogramming were optimized to maximize plasmid uptake and cell survival."
- "Antimicrobial resistance in mycobacteria arises from a complex interplay of intrinsic and acquired mechanisms that collectively limit the efficacy of current therapeutic options."
- "Current experimental and clinical evidence suggests that pathogenesis reflects both cholesterol insufficiency and sterol-mediated toxicity"
- "The adult heart replaces cardiomyocytes at approximately 1% per year in young adults, declining to about 0.45% per year with ageing"
- "Curative approaches have advanced substantially through gene addition, gene editing, and base editing technologies."
- "This study establishes genetic mosaicism as an important feature of Cas9-mediated gene editing in Daphnia."
- "These methods provide an experimental approach with translational potential for externally controlled therapeutic cargo release."
- "This system combines two distinct morphogenetic regulators, the wheat GRF4-GIF1 chimera and the maize BABY BOOM (BBM) transcription factor (hence the name "GGB") with a modified QuickCorn protocol, enabling regeneration of transformed maize plantlets in c. 2 months with an efficiency 7-fold higher than when compared to either morphogenic factor used in isolation."
- "Doxycycline (Dox)-inducible overexpression (OE) of Pcgf5 reduced the double-positive (DP) 2C-like population."
- "Public opinion strongly supports using genetic editing to treat life-threatening diseases in adults and embryos (63.2% and 73.6%, respectively)."