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^*

^*Corresponding author for this work

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

109 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

Bibliographical note

Publisher Copyright:
© 2021, The Author(s).

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

Cite this

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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",

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year = "2021",

month = dec,

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

language = "English",

volume = "22",

journal = "Genome Biology",

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T1 - Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor

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

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.

KW - Adamts20

KW - Chromatin-modulating peptides

KW - Dwarf phenotype

KW - Germline transmission

KW - Igf2

KW - Mouse cells and embryos

KW - Prime editor

KW - Proximal dead sgRNA

UR - https://www.scopus.com/pages/publications/85107198784

U2 - 10.1186/s13059-021-02389-w

DO - 10.1186/s13059-021-02389-w

M3 - Article

C2 - 34082781

AN - SCOPUS:85107198784

SN - 1474-7596

VL - 22

JO - Genome Biology

JF - Genome Biology

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M1 - 170

ER -

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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