APOBEC3 induces mutations during repair of CRISPR–Cas9-generated DNA breaks
| Autor: | Jia Wei, Jianying Wang, Lei Yan, Wei Li, Xingxu Huang, Bian Hu, Min Zhuang, Bei Yang, Liqun Lei, Jimin Gao, Hongquan Chen, Wei Xue, Jia Chen, Yiqiang Cui, Lijie Wang, Bin Shen, Jiahao Sha, Wanjing Shang, Li Yang |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
DNA Repair DNA repair Oligonucleotides Deamination DNA Single-Stranded Cytidine medicine.disease_cause Cytosine Deaminase 03 medical and health sciences chemistry.chemical_compound Cytidine deamination INDEL Mutation Structural Biology Cytidine Deaminase medicine Humans DNA Breaks Double-Stranded APOBEC Deaminases RNA Small Interfering Molecular Biology Mutation Mutagenesis Recombinational DNA Repair food and beverages Sequence Analysis DNA Cytidine deaminase Cell biology genomic DNA HEK293 Cells 030104 developmental biology chemistry CRISPR-Cas Systems DNA HeLa Cells |
| Zdroj: | Nature Structural & Molecular Biology. 25:45-52 |
| ISSN: | 1545-9985 1545-9993 |
| DOI: | 10.1038/s41594-017-0004-6 |
| Popis: | The APOBEC-AID family of cytidine deaminase prefers single-stranded nucleic acids for cytidine-to-uridine deamination. Single-stranded nucleic acids are commonly involved in the DNA repair system for breaks generated by CRISPR-Cas9. Here, we show in human cells that APOBEC3 can trigger cytidine deamination of single-stranded oligodeoxynucleotides, which ultimately results in base substitution mutations in genomic DNA through homology-directed repair (HDR) of Cas9-generated double-strand breaks. In addition, the APOBEC3-catalyzed deamination in genomic single-stranded DNA formed during the repair of Cas9 nickase-generated single-strand breaks in human cells can be further processed to yield mutations mainly involving insertions or deletions (indels). Both APOBEC3-mediated deamination and DNA-repair proteins play important roles in the generation of these indels. Therefore, optimizing conditions for the repair of CRISPR-Cas9-generated DNA breaks, such as using double-stranded donors in HDR or temporarily suppressing endogenous APOBEC3s, can repress these unwanted mutations in genomic DNA. |
| Databáze: | OpenAIRE |
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