Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor

Soo Ji Park; Tae Yeong Jeong; Seung Kyun Shin; Da Eun Yoon; Soo Yeon Lim; Sol Pin Kim; Jungmin Choi; Hyunji Lee; Jeong Im Hong; Jinhee Ahn; Je Kyung Seong; Kyoungmi Kim

doi:10.1186/s13059-021-02389-w

Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor

Soo Ji Park, Tae Yeong Jeong, Seung Kyun Shin, Da Eun Yoon, Soo Yeon Lim, Sol Pin Kim, Jungmin Choi, Hyunji Lee, Jeong Im Hong, Jinhee Ahn, Je Kyung Seong, Kyoungmi Kim

Department of Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

43 Citations (Scopus)

Abstract

Prime editors, novel genome-editing tools consisting of a CRISPR-Cas9 nickase and an engineered reverse transcriptase, can induce targeted mutagenesis. Nevertheless, much effort is required to optimize and improve the efficiency of prime-editing. Herein, we introduce two strategies to improve the editing efficiency using proximal dead sgRNA and chromatin-modulating peptides. We used enhanced prime-editing to generate Igf2 mutant mice with editing frequencies of up to 47% and observed germline transmission, no off-target effects, and a dwarf phenotype. This improved prime-editing method can be efficiently applied to cell research and to generate mouse models.

Original language	English
Article number	170
Journal	Genome Biology
Volume	22
Issue number	1
DOIs	https://doi.org/10.1186/s13059-021-02389-w
Publication status	Published - 2021 Dec

Keywords

Adamts20
Chromatin-modulating peptides
Dwarf phenotype
Germline transmission
Igf2
Mouse cells and embryos
Prime editor
Proximal dead sgRNA

ASJC Scopus subject areas

Ecology, Evolution, Behavior and Systematics
Genetics
Cell Biology

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10.1186/s13059-021-02389-w

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title = "Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor",

abstract = "Prime editors, novel genome-editing tools consisting of a CRISPR-Cas9 nickase and an engineered reverse transcriptase, can induce targeted mutagenesis. Nevertheless, much effort is required to optimize and improve the efficiency of prime-editing. Herein, we introduce two strategies to improve the editing efficiency using proximal dead sgRNA and chromatin-modulating peptides. We used enhanced prime-editing to generate Igf2 mutant mice with editing frequencies of up to 47% and observed germline transmission, no off-target effects, and a dwarf phenotype. This improved prime-editing method can be efficiently applied to cell research and to generate mouse models.",

keywords = "Adamts20, Chromatin-modulating peptides, Dwarf phenotype, Germline transmission, Igf2, Mouse cells and embryos, Prime editor, Proximal dead sgRNA",

author = "Park, {Soo Ji} and Jeong, {Tae Yeong} and Shin, {Seung Kyun} and Yoon, {Da Eun} and Lim, {Soo Yeon} and Kim, {Sol Pin} and Jungmin Choi and Hyunji Lee and Hong, {Jeong Im} and Jinhee Ahn and Seong, {Je Kyung} and Kyoungmi Kim",

note = "Funding Information: This study was supported by the Chung Yang, Cha Young Sun, & Jang Hi Joo Memorial Fund. This study was also supported by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) of Korea (Korea Mouse Phenotyping Project, NRF-2013M3A9D5072550, NRF-2020M3A9D5A01082439, NRF-2019R1A2C2087198, and NRF- 2019M3A9H1103792). Publisher Copyright: {\textcopyright} 2021, The Author(s).",

year = "2021",

month = dec,

doi = "10.1186/s13059-021-02389-w",

language = "English",

volume = "22",

journal = "Genome Biology",

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number = "1",

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AU - Park, Soo Ji

AU - Jeong, Tae Yeong

AU - Shin, Seung Kyun

AU - Yoon, Da Eun

AU - Lim, Soo Yeon

AU - Kim, Sol Pin

AU - Choi, Jungmin

AU - Lee, Hyunji

AU - Hong, Jeong Im

AU - Ahn, Jinhee

AU - Seong, Je Kyung

AU - Kim, Kyoungmi

N1 - Funding Information: This study was supported by the Chung Yang, Cha Young Sun, & Jang Hi Joo Memorial Fund. This study was also supported by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) of Korea (Korea Mouse Phenotyping Project, NRF-2013M3A9D5072550, NRF-2020M3A9D5A01082439, NRF-2019R1A2C2087198, and NRF- 2019M3A9H1103792). Publisher Copyright: © 2021, The Author(s).

PY - 2021/12

Y1 - 2021/12

N2 - Prime editors, novel genome-editing tools consisting of a CRISPR-Cas9 nickase and an engineered reverse transcriptase, can induce targeted mutagenesis. Nevertheless, much effort is required to optimize and improve the efficiency of prime-editing. Herein, we introduce two strategies to improve the editing efficiency using proximal dead sgRNA and chromatin-modulating peptides. We used enhanced prime-editing to generate Igf2 mutant mice with editing frequencies of up to 47% and observed germline transmission, no off-target effects, and a dwarf phenotype. This improved prime-editing method can be efficiently applied to cell research and to generate mouse models.

AB - Prime editors, novel genome-editing tools consisting of a CRISPR-Cas9 nickase and an engineered reverse transcriptase, can induce targeted mutagenesis. Nevertheless, much effort is required to optimize and improve the efficiency of prime-editing. Herein, we introduce two strategies to improve the editing efficiency using proximal dead sgRNA and chromatin-modulating peptides. We used enhanced prime-editing to generate Igf2 mutant mice with editing frequencies of up to 47% and observed germline transmission, no off-target effects, and a dwarf phenotype. This improved prime-editing method can be efficiently applied to cell research and to generate mouse models.

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KW - Chromatin-modulating peptides

KW - Dwarf phenotype

KW - Germline transmission

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SN - 1474-7596

VL - 22

JO - Genome Biology

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Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor

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Keywords

ASJC Scopus subject areas

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