Targeted genome editing with a DNA-dependent DNA polymerase and exogenous DNA-containing templates

被引：19
|
作者：
Liu, Bin [1 ]
Dong, Xiaolong [1 ,6 ]
Zheng, Chunwei [1 ]
Keener, David [1 ]
Chen, Zexiang [1 ]
Cheng, Haoyang [1 ]
Watts, Jonathan K. [1 ,2 ,3 ]
Xue, Wen [1 ,2 ,4 ,5 ]
Sontheimer, Erik J. [1 ,2 ,3 ,5 ]
机构：
[1] Univ Massachusetts, Chan Med Sch, RNA Therapeut Inst, Worcester, MA 01605 USA
[2] Univ Massachusetts, Chan Med Sch, Li Weibo Inst Rare Dis Res, Worcester, MA 01605 USA
[3] Univ Massachusetts, Chan Med Sch, Dept Biochem & Mol Biotechnol, Worcester, MA 01605 USA
[4] Univ Massachusetts, Chan Med Sch, Dept Mol Cell & Canc Biol, Worcester, MA 01605 USA
[5] Univ Massachusetts, Chan Med Sch, Dept Mol Med, Worcester, MA 01605 USA
[6] Tessera Therapeut, Somerville, MA USA
来源：
NATURE BIOTECHNOLOGY | 2024年 / 42卷 / 07期
关键词：
FIDELITY;
D O I：
10.1038/s41587-023-01947-w
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Reverse transcriptases, used in prime editing systems, exhibit lower fidelity, processivity and dNTP affinity than many DNA-dependent DNA polymerases. We report that a DNA-dependent DNA polymerase (phi29), untethered from Cas9, enables editing from a synthetic, end-stabilized DNA-containing template at up to 60% efficiency in human cells. Compared to prime editing, DNA polymerase editing avoids autoinhibitory intramolecular base pairing of the template, facilitates template synthesis and supports larger insertions (>100 nucleotides).
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页码：1039 / 1045
页数：21
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