ArticleGenome research2018
Precise and efficient nucleotide substitution near genomic nick via noncanonical homology-directed repair.
Article in Genome research, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed.
- Production and Storage of High-Capacity Adenoviral Vectors for the Delivery of Advanced CRISPR Systems.Methods in molecular biology (Clifton, N.J.) · 2027Article
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- On RNA-programmable gene modulation as a versatile set of principles targeting muscular dystrophies.Molecular therapy : the journal of the American Society of Gene Therapy · 2024Review
- A combined transcriptomic and physiological approach to understanding the adaptive mechanisms to cope with oxidative stress inMicrobiology spectrum · 2023Article
- Inducing multiple nicks promotes interhomolog homologous recombination to correct heterozygous mutations in somatic cells.Nature communications · 2023Article
- High-efficiency and multilocus targeted integration in CHO cells using CRISPR-mediated donor nicking and DNA repair inhibitors.Biotechnology and bioengineering · 2023Article
- Precise homology-directed installation of large genomic edits in human cells with cleaving and nicking high-specificity Cas9 variants.Nucleic acids research · 2023Article
- Flow cytometry-based quantification of genome editing efficiency in human cell lines using the L1CAM gene.PloS one · 2023Article
- In search of an ideal template for therapeutic genome editing: A review of current developments for structure optimization.Frontiers in genome editing · 2023Review
- Correction of a CD55 mutation to quantify the efficiency of targeted knock-in via flow cytometry.Molecular biology reports · 2022Article
- Precise CRISPR-Cas-mediated gene repair with minimal off-target and unintended on-target mutations in human hematopoietic stem cells.Science advances · 2022Article
- CRISPR-based genome editing through the lens of DNA repair.Molecular cell · 2022Review
- Flow cytometry-based quantification of targeted knock-in events in human cell lines using a GPI-anchor biosynthesis gene PIGP.Bioscience reports · 2021Article
- Experimental strategies to achieve efficient targeted knock-in via tandem paired nicking.Scientific reports · 2021Article
- Broadening the reach and investigating the potential of prime editors through fully viral gene-deleted adenoviral vector delivery.Nucleic acids research · 2021Article
- Different DNA repair pathways are involved in single-strand break-induced genomic changes in plants.The Plant cell · 2021Article
- Next-Generation CRISPR Technologies and Their Applications in Gene and Cell Therapy.Trends in biotechnology · 2021Review
- HeterozygousScience immunology · 2021Observational
- Use of single guided Cas9 nickase to facilitate precise and efficient genome editing in human iPSCs.Scientific reports · 2021Article
- Precise and broad scope genome editing based on high-specificity Cas9 nickases.Nucleic acids research · 2021Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR/Cas9, which generates DNA double-strand breaks (DSBs) at target loci, is a powerful tool for editing genomes when codelivered with a donor DNA template. However, DSBs, which are the most deleterious type of DNA damage, often result in unintended nucleotide insertions/deletions (indels) via mutagenic nonhomologous end joining. We developed a strategy for precise gene editing that does not generate DSBs. We show that a combination of single nicks in the target gene and donor plasmid (SNGD) using Cas9D10A nickase promotes efficient nucleotide substitution by gene editing. Nicking the target gene alone did not facilitate efficient gene editing. However, an additional nick in the donor plasmid backbone markedly improved the gene-editing efficiency. SNGD-mediated gene editing led to a markedly lower indel frequency than that by the DSB-mediated approach. We also show that SNGD promotes gene editing at endogenous loci in human cells. Mechanistically, SNGD-mediated gene editing requires long-sequence homology between the target gene and repair template, but does not require CtIP, RAD51, or RAD52. Thus, it is considered that noncanonical homology-directed repair regulates the SNGD-mediated gene editing. In summary, SNGD promotes precise and efficient gene editing and may be a promising strategy for the development of a novel gene therapy approach.
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