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同源修复模板DNA的链间交联增强了人类细胞中的基因编辑。

Interstrand crosslinking of homologous repair template DNA enhances gene editing in human cells.

作者信息

Ghasemi Hannah I, Bacal Julien, Yoon Amanda C, Tavasoli Katherine U, Cruz Carmen, Vu Jonathan T, Gardner Brooke M, Richardson Chris D

机构信息

Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA, USA.

出版信息

Nat Biotechnol. 2023 Oct;41(10):1398-1404. doi: 10.1038/s41587-022-01654-y. Epub 2023 Feb 27.

DOI:10.1038/s41587-022-01654-y
PMID:36849829
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10460463/
Abstract

We describe a strategy to boost the efficiency of gene editing via homology-directed repair (HDR) by covalently modifying the template DNA with interstrand crosslinks. Crosslinked templates (xHDRTs) increase Cas9-mediated editing efficiencies by up to fivefold in K562, HEK293T, U2OS, iPS and primary T cells. Increased editing from xHDRTs is driven by events on the template molecule and requires ataxia telangiectasia and Rad3-related (ATR) kinase and components of the Fanconi anemia pathway.

摘要

我们描述了一种通过用链间交联共价修饰模板DNA来提高同源定向修复（HDR）基因编辑效率的策略。交联模板（xHDRTs）在K562、HEK293T、U2OS、诱导多能干细胞（iPS）和原代T细胞中可将Cas9介导的编辑效率提高多达五倍。xHDRTs编辑效率的提高是由模板分子上的事件驱动的，并且需要共济失调毛细血管扩张症和Rad3相关（ATR）激酶以及范可尼贫血途径的成分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/52af26147921/41587_2022_1654_Fig11_ESM.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/c664acdc8085/41587_2022_1654_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/3d3559e75048/41587_2022_1654_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/69fea1fc9056/41587_2022_1654_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/bc8028fdf05f/41587_2022_1654_Fig4_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/cd9a6d08f390/41587_2022_1654_Fig5_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/07c4a64dcff8/41587_2022_1654_Fig6_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/118e8062e0fa/41587_2022_1654_Fig7_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/18c134d2f120/41587_2022_1654_Fig8_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/38d4c2cf4e7b/41587_2022_1654_Fig9_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/77efcdc92ff8/41587_2022_1654_Fig10_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c415/10567570/52af26147921/41587_2022_1654_Fig11_ESM.jpg

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