CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression

The efficiency of homology-directed repair (HDR) plays a crucial role in the development of animal models and gene therapy. We demonstrate that microhomology-mediated end-joining (MMEJ) constitutes a substantial proportion of DNA repair during CRISPR-mediated gene editing. Using CasRx to downregulat...

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Autores principales: Chen, Hongyu, Liu, Xingchen, Li, Lanxin, Tan, Qingtong, Li, Shiyan, Li, Li, Li, Chunyang, Fu, Jiqiang, Lu, Yong, Wang, Yan, Sun, Yidi, Luo, Zhen-Ge, Lu, Zongyang, Sun, Qiang, Liu, Zhen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10288798/
https://www.ncbi.nlm.nih.gov/pubmed/37353834
http://dx.doi.org/10.1186/s13059-023-02987-w
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author Chen, Hongyu
Liu, Xingchen
Li, Lanxin
Tan, Qingtong
Li, Shiyan
Li, Li
Li, Chunyang
Fu, Jiqiang
Lu, Yong
Wang, Yan
Sun, Yidi
Luo, Zhen-Ge
Lu, Zongyang
Sun, Qiang
Liu, Zhen
author_facet Chen, Hongyu
Liu, Xingchen
Li, Lanxin
Tan, Qingtong
Li, Shiyan
Li, Li
Li, Chunyang
Fu, Jiqiang
Lu, Yong
Wang, Yan
Sun, Yidi
Luo, Zhen-Ge
Lu, Zongyang
Sun, Qiang
Liu, Zhen
author_sort Chen, Hongyu
collection PubMed
description The efficiency of homology-directed repair (HDR) plays a crucial role in the development of animal models and gene therapy. We demonstrate that microhomology-mediated end-joining (MMEJ) constitutes a substantial proportion of DNA repair during CRISPR-mediated gene editing. Using CasRx to downregulate a key MMEJ factor, Polymerase Q (Polq), we improve the targeted integration efficiency of linearized DNA fragments and single-strand oligonucleotides (ssODN) in mouse embryos and offspring. CasRX-assisted targeted integration (CATI) also leads to substantial improvements in HDR efficiency during the CRISPR/Cas9 editing of monkey embryos. We present a promising tool for generating monkey models and developing gene therapies for clinical trials. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-023-02987-w.
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spelling pubmed-102887982023-06-24 CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression Chen, Hongyu Liu, Xingchen Li, Lanxin Tan, Qingtong Li, Shiyan Li, Li Li, Chunyang Fu, Jiqiang Lu, Yong Wang, Yan Sun, Yidi Luo, Zhen-Ge Lu, Zongyang Sun, Qiang Liu, Zhen Genome Biol Method The efficiency of homology-directed repair (HDR) plays a crucial role in the development of animal models and gene therapy. We demonstrate that microhomology-mediated end-joining (MMEJ) constitutes a substantial proportion of DNA repair during CRISPR-mediated gene editing. Using CasRx to downregulate a key MMEJ factor, Polymerase Q (Polq), we improve the targeted integration efficiency of linearized DNA fragments and single-strand oligonucleotides (ssODN) in mouse embryos and offspring. CasRX-assisted targeted integration (CATI) also leads to substantial improvements in HDR efficiency during the CRISPR/Cas9 editing of monkey embryos. We present a promising tool for generating monkey models and developing gene therapies for clinical trials. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-023-02987-w. BioMed Central 2023-06-23 /pmc/articles/PMC10288798/ /pubmed/37353834 http://dx.doi.org/10.1186/s13059-023-02987-w Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Method
Chen, Hongyu
Liu, Xingchen
Li, Lanxin
Tan, Qingtong
Li, Shiyan
Li, Li
Li, Chunyang
Fu, Jiqiang
Lu, Yong
Wang, Yan
Sun, Yidi
Luo, Zhen-Ge
Lu, Zongyang
Sun, Qiang
Liu, Zhen
CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression
title CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression
title_full CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression
title_fullStr CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression
title_full_unstemmed CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression
title_short CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression
title_sort cati: an efficient gene integration method for rodent and primate embryos by mmej suppression
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10288798/
https://www.ncbi.nlm.nih.gov/pubmed/37353834
http://dx.doi.org/10.1186/s13059-023-02987-w
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