ArticleNucleic acids research2018
Highly efficient genome editing via CRISPR-Cas9 in human pluripotent stem cells is achieved by transient BCL-XL overexpression.
Article in Nucleic acids research, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 77 papers.
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Who cites it
77 citing papers in PubMed.
- Tgfbr3 is required for the long-term maintenance of self-renewal potential in murine hematopoietic stem cells.Blood science (Baltimore, Md.) · 2026Article
- CRISPR/Cas9-based repair of a heterozygous HNF1A mutation in patient-derived hiPSCs.Human genetics · 2026Article
- Arrayed dual-gRNA CRISPR screening platform forMolecular therapy. Advances · 2026Article
- Universal conditional knockout approach to multiple species, from fish to human induced pluripotent stem cells.Nucleic acids research · 2026Article
- CRISPR and the Future of Cardiac Disease Therapy: A New Genetic Frontier.International journal of molecular sciences · 2026Review
- Omics-aided design genome editing strategy for challenging human immortalized cell models.PloS one · 2026Article
- Programmable multistep CRISPR gene activation via control of RNA polymerase III termination.Science advances · 2025Article
- G2B: an optimized lentiviral vector with enhanced titer and β-globin expression for improved β-thalassemia gene therapy.Blood science (Baltimore, Md.) · 2025Article
- Improving Gene Knock-In Efficiencies in Sheep Primary Cells Using a CRISPR/Cas9-Gal4 System.Current issues in molecular biology · 2025Article
- Human Genome REWRITE for Off-the-Shelf Stem Cells Reveals an "Epigenetic Ghost".bioRxiv : the preprint server for biology · 2025Article
- Focused-ultrasound-mediated CRISPR-Cas9 gene editing in human induced pluripotent stem cells.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Article
- CRISPR/Cas9-Based Protocol for Precise Genome Editing in Induced Pluripotent Stem Cells.Bio-protocol · 2024Article
- A p21 reporter iPSC line for evaluating CRISPR-Cas9 and vector-induced stress responses.Stem cells (Dayton, Ohio) · 2024Article
- Genetically engineered human induced pluripotent stem cells for the production of brain-targeting extracellular vesicles.Stem cell research & therapy · 2024Article
- Article
- OliTag-seq enhances in cellulo detection of CRISPR-Cas9 off-targets.Communications biology · 2024Article
- Therapeutic validation of MMR-associated genetic modifiers in a human ex vivo model of Huntington disease.American journal of human genetics · 2024Article
- A high efficiency precision genome editing method with CRISPR in iPSCs.Scientific reports · 2024Article
- Comprehensive review of CRISPR-based gene editing: mechanisms, challenges, and applications in cancer therapy.Molecular cancer · 2024Review
- Correction of Griscelli Syndrome Type 2 causing mutations in theTurkish journal of biology = Turk biyoloji dergisi · 2024Article
17 more citing papers are in PubMed but not listed here.
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20 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Genome editing of human induced pluripotent stem cells (iPSCs) is instrumental for functional genomics, disease modeling, and regenerative medicine. However, low editing efficiency has hampered the applications of CRISPR-Cas9 technology in creating knockin (KI) or knockout (KO) iPSC lines, which is largely due to massive cell death after electroporation with editing plasmids. Here, we report that the transient delivery of BCL-XL increases iPSC survival by ∼10-fold after plasmid transfection, leading to a 20- to 100-fold increase in homology-directed repair (HDR) KI efficiency and a 5-fold increase in non-homologous end joining (NHEJ) KO efficiency. Treatment with a BCL inhibitor ABT-263 further improves HDR efficiency by 70% and KO efficiency by 40%. The increased genome editing efficiency is attributed to higher expressions of Cas9 and sgRNA in surviving cells after electroporation. HDR or NHEJ efficiency reaches 95% with dual editing followed by selection of cells with HDR insertion of a selective gene. Moreover, KO efficiency of 100% can be achieved in a bulk population of cells with biallelic HDR KO followed by double selection, abrogating the necessity for single cell cloning. Taken together, these simple yet highly efficient editing strategies provide useful tools for applications ranging from manipulating human iPSC genomes to creating gene-modified animal models.
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