Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds

CRISPR-Cas12a/Cpf1, a single RNA-guided endonuclease system, provides a promising tool for genome engineering. However, only three Cas12a orthologs have been employed for mammalian genome editing, and the editing efficiency as well as targeting coverage still requires improvements. Here, we harness...

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Autores principales: Teng, Fei, Li, Jing, Cui, Tongtong, Xu, Kai, Guo, Lu, Gao, Qingqin, Feng, Guihai, Chen, Chuanyuan, Han, Dali, Zhou, Qi, Li, Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Short Report
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362571/
https://www.ncbi.nlm.nih.gov/pubmed/30717767
http://dx.doi.org/10.1186/s13059-019-1620-8
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author Teng, Fei
Li, Jing
Cui, Tongtong
Xu, Kai
Guo, Lu
Gao, Qingqin
Feng, Guihai
Chen, Chuanyuan
Han, Dali
Zhou, Qi
Li, Wei
author_facet Teng, Fei
Li, Jing
Cui, Tongtong
Xu, Kai
Guo, Lu
Gao, Qingqin
Feng, Guihai
Chen, Chuanyuan
Han, Dali
Zhou, Qi
Li, Wei
author_sort Teng, Fei
collection PubMed
description CRISPR-Cas12a/Cpf1, a single RNA-guided endonuclease system, provides a promising tool for genome engineering. However, only three Cas12a orthologs have been employed for mammalian genome editing, and the editing efficiency as well as targeting coverage still requires improvements. Here, we harness six novel Cas12a orthologs for genome editing in human and mouse cells, some of which utilize simple protospacer adjacent motifs (PAMs) that remarkably increase the targeting range in the genomes. Moreover, we identify optimized CRISPR RNA (crRNA) scaffolds that can increase the genome editing efficiency of Cas12a. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13059-019-1620-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-63625712019-02-14 Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds Teng, Fei Li, Jing Cui, Tongtong Xu, Kai Guo, Lu Gao, Qingqin Feng, Guihai Chen, Chuanyuan Han, Dali Zhou, Qi Li, Wei Genome Biol Short Report CRISPR-Cas12a/Cpf1, a single RNA-guided endonuclease system, provides a promising tool for genome engineering. However, only three Cas12a orthologs have been employed for mammalian genome editing, and the editing efficiency as well as targeting coverage still requires improvements. Here, we harness six novel Cas12a orthologs for genome editing in human and mouse cells, some of which utilize simple protospacer adjacent motifs (PAMs) that remarkably increase the targeting range in the genomes. Moreover, we identify optimized CRISPR RNA (crRNA) scaffolds that can increase the genome editing efficiency of Cas12a. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13059-019-1620-8) contains supplementary material, which is available to authorized users. BioMed Central 2019-02-05 /pmc/articles/PMC6362571/ /pubmed/30717767 http://dx.doi.org/10.1186/s13059-019-1620-8 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Short Report
Teng, Fei
Li, Jing
Cui, Tongtong
Xu, Kai
Guo, Lu
Gao, Qingqin
Feng, Guihai
Chen, Chuanyuan
Han, Dali
Zhou, Qi
Li, Wei
Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds
title Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds
title_full Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds
title_fullStr Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds
title_full_unstemmed Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds
title_short Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds
title_sort enhanced mammalian genome editing by new cas12a orthologs with optimized crrna scaffolds
topic Short Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362571/
https://www.ncbi.nlm.nih.gov/pubmed/30717767
http://dx.doi.org/10.1186/s13059-019-1620-8
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