ArticleNucleic acids research2015
Multi-kilobase homozygous targeted gene replacement in human induced pluripotent stem cells.
Article in Nucleic acids research, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 87 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
87 citing papers in PubMed, 170 citations in OpenAlex.
- Toward transgenic multicellular parasites: CRISPR-Cas12a germline editing ofScience advances · 2026Article
- Improving Gene Knock-In Efficiencies in Sheep Primary Cells Using a CRISPR/Cas9-Gal4 System.Current issues in molecular biology · 2025Article
- KDM6A facilitates Xist upregulation at the onset of X inactivation.Biology of sex differences · 2025Article
- In vivo affinity maturation of mouse B cells reprogrammed to express human antibodies.Nature biomedical engineering · 2024Article
- Recent advances in CRISPR-Cas9-based genome insertion technologies.Molecular therapy. Nucleic acids · 2024Review
- Outlook on genome editing application to cattle.Journal of veterinary science · 2024Review
- Making gene editing accessible in resource limited environments: recommendations to guide a first-time user.Frontiers in genome editing · 2024Review
- A structural peculiarity of Antarctic fish IgM drives the generation of an engineered mAb by CRISPR/Cas9.Frontiers in bioengineering and biotechnology · 2024Article
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- Advances in CRISPR/Cas gene therapy for inborn errors of immunity.Frontiers in immunology · 2023Review
- Transcription-coupled donor DNA expression increases homologous recombination for efficient genome editing.Nucleic acids research · 2022Article
- Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells.Nature communications · 2022Article
- STRAIGHT-IN enables high-throughput targeting of large DNA payloads in human pluripotent stem cells.Cell reports methods · 2022Article
- Sex-biased and parental allele-specific gene regulation by KDM6A.Biology of sex differences · 2022Article
- Large-scale genome editing based on high-capacity adenovectors and CRISPR-Cas9 nucleases rescues full-length dystrophin synthesis in DMD muscle cells.Nucleic acids research · 2022Article
- Biallelic and gene-wide genomic substitution for endogenous intron and retroelement mutagenesis in human cells.Nature communications · 2022Article
- An update on precision genome editing by homology-directed repair in plants.Plant physiology · 2022Article
- Applications of and considerations for using CRISPR-Cas9-mediated gene conversion systems in rodents.Nature protocols · 2022Review
- Inherent hepatocytic heterogeneity determines expression and retention of editedProceedings of the National Academy of Sciences of the United States of America · 2021Article
- Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells.Scientific reports · 2021Article
27 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Sequence-specific nucleases such as TALEN and the CRISPR/Cas9 system have so far been used to disrupt, correct or insert transgenes at precise locations in mammalian genomes. We demonstrate efficient 'knock-in' targeted replacement of multi-kilobase genes in human induced pluripotent stem cells (iPSC). Using a model system replacing endogenous human genes with their mouse counterpart, we performed a comprehensive study of targeting vector design parameters for homologous recombination. A 2.7 kilobase (kb) homozygous gene replacement was achieved in up to 11% of iPSC without selection. The optimal homology arm length was around 2 kb, with homology length being especially critical on the arm not adjacent to the cut site. Homologous sequence inside the cut sites was detrimental to targeting efficiency, consistent with a synthesis-dependent strand annealing (SDSA) mechanism. Using two nuclease sites, we observed a high degree of gene excisions and inversions, which sometimes occurred more frequently than indel mutations. While homozygous deletions of 86 kb were achieved with up to 8% frequency, deletion frequencies were not solely a function of nuclease activity and deletion size. Our results analyzing the optimal parameters for targeting vector design will inform future gene targeting efforts involving multi-kilobase gene segments, particularly in human iPSC.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.