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).

引用

页码：1039 / 1045

页数：21

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