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Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells
Cpf1, an RNA-guided DNA endonuclease that belongs to a new class II CRISPR system, has recently been harnessed for genome editing. Herein, we report an RNase-resistant caged truncated pre-tRNA-like crRNA (catRNA) that confers precise and efficient gene editing with the Lachnospiraceae bacterium Cpf1...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6037731/ https://www.ncbi.nlm.nih.gov/pubmed/30002872 http://dx.doi.org/10.1038/s41421-018-0035-0 |
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author | Zhang, Xuhua Xu, Linping Fan, Ruihua Gao, Quanli Song, Yunfeng Lyu, Xiaodong Ren, Jiangtao Song, Yongping |
author_facet | Zhang, Xuhua Xu, Linping Fan, Ruihua Gao, Quanli Song, Yunfeng Lyu, Xiaodong Ren, Jiangtao Song, Yongping |
author_sort | Zhang, Xuhua |
collection | PubMed |
description | Cpf1, an RNA-guided DNA endonuclease that belongs to a new class II CRISPR system, has recently been harnessed for genome editing. Herein, we report an RNase-resistant caged truncated pre-tRNA-like crRNA (catRNA) that confers precise and efficient gene editing with the Lachnospiraceae bacterium Cpf1 (LbCpf1) and enables the reprogramming of catalytically dead LbCpf1 (dCpf1) lacking DNA endonuclease activity into a transcriptional modulator. Specific gene knock-outs and knock-ins were increased 3.2-fold and 4.3-fold, respectively, with catRNA compared to that induced by conventional crRNA. A much higher augmentation of gene disruption (up to 37-fold) was observed when electroporation was used. We report herein that catRNA enables efficient gene activation with dCpf1 activators. Our study reveals the potential of catRNA and a versatile application of the CRISPR/Cpf1 system, establishing a simple approach for selective gene perturbation in mammalian cells. |
format | Online Article Text |
id | pubmed-6037731 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-60377312018-07-12 Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells Zhang, Xuhua Xu, Linping Fan, Ruihua Gao, Quanli Song, Yunfeng Lyu, Xiaodong Ren, Jiangtao Song, Yongping Cell Discov Article Cpf1, an RNA-guided DNA endonuclease that belongs to a new class II CRISPR system, has recently been harnessed for genome editing. Herein, we report an RNase-resistant caged truncated pre-tRNA-like crRNA (catRNA) that confers precise and efficient gene editing with the Lachnospiraceae bacterium Cpf1 (LbCpf1) and enables the reprogramming of catalytically dead LbCpf1 (dCpf1) lacking DNA endonuclease activity into a transcriptional modulator. Specific gene knock-outs and knock-ins were increased 3.2-fold and 4.3-fold, respectively, with catRNA compared to that induced by conventional crRNA. A much higher augmentation of gene disruption (up to 37-fold) was observed when electroporation was used. We report herein that catRNA enables efficient gene activation with dCpf1 activators. Our study reveals the potential of catRNA and a versatile application of the CRISPR/Cpf1 system, establishing a simple approach for selective gene perturbation in mammalian cells. Nature Publishing Group UK 2018-07-10 /pmc/articles/PMC6037731/ /pubmed/30002872 http://dx.doi.org/10.1038/s41421-018-0035-0 Text en © The Author(s) 2018 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Zhang, Xuhua Xu, Linping Fan, Ruihua Gao, Quanli Song, Yunfeng Lyu, Xiaodong Ren, Jiangtao Song, Yongping Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells |
title | Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells |
title_full | Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells |
title_fullStr | Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells |
title_full_unstemmed | Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells |
title_short | Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells |
title_sort | genetic editing and interrogation with cpf1 and caged truncated pre-trna-like crrna in mammalian cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6037731/ https://www.ncbi.nlm.nih.gov/pubmed/30002872 http://dx.doi.org/10.1038/s41421-018-0035-0 |
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