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Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass
An ideal rAAV gene editing system not only effectively edits genes at specific site, but also prevents the spread of the virus from occurring off-target or carcinogenic risks. This is important for gene editing research at specific site in vivo. We report a single rAAV containing SaCas9 and guide RN...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8510912/ https://www.ncbi.nlm.nih.gov/pubmed/34590222 http://dx.doi.org/10.1007/s10529-021-03183-1 |
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author | Weng, Shaoting Zhao, Yitian Yu, Changhong Wang, Xiaofan Xiao, Xuehan Han, Liqiang Zhang, Kunpeng Wang, Jiang Yang, Guoyu |
author_facet | Weng, Shaoting Zhao, Yitian Yu, Changhong Wang, Xiaofan Xiao, Xuehan Han, Liqiang Zhang, Kunpeng Wang, Jiang Yang, Guoyu |
author_sort | Weng, Shaoting |
collection | PubMed |
description | An ideal rAAV gene editing system not only effectively edits genes at specific site, but also prevents the spread of the virus from occurring off-target or carcinogenic risks. This is important for gene editing research at specific site in vivo. We report a single rAAV containing SaCas9 and guide RNAs under the control of subtle EF1a and tRNA promoters. The capacity of rAAV was compressed, and the editing efficiency was similar to that of the classical Cas9 system in vitro and in vivo. And we inserted the sequence of the green fluorescent protein eGFP into rAAV. The number of cells infected with the rAAV and the region in which the rAAV spreads were known by the fluorescent expression of eGFP in cells. In addition, we demonstrated that myostatin gene in the thigh muscles of C57BL/10 mice was knocked out by the rAAV9-SaCas9 system to make muscle mass increased obviously. The protein eGFP into rAAV has significant implications for our indirect analysis of the editing efficiency of SaCas9 in the genome of the target tissue and reduces the harm caused by off-target editing and prevents other tissue mutations. The rAAV system has substantial potential in improving muscle mass and preventing muscle atrophy. |
format | Online Article Text |
id | pubmed-8510912 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-85109122021-10-19 Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass Weng, Shaoting Zhao, Yitian Yu, Changhong Wang, Xiaofan Xiao, Xuehan Han, Liqiang Zhang, Kunpeng Wang, Jiang Yang, Guoyu Biotechnol Lett Original Research Paper An ideal rAAV gene editing system not only effectively edits genes at specific site, but also prevents the spread of the virus from occurring off-target or carcinogenic risks. This is important for gene editing research at specific site in vivo. We report a single rAAV containing SaCas9 and guide RNAs under the control of subtle EF1a and tRNA promoters. The capacity of rAAV was compressed, and the editing efficiency was similar to that of the classical Cas9 system in vitro and in vivo. And we inserted the sequence of the green fluorescent protein eGFP into rAAV. The number of cells infected with the rAAV and the region in which the rAAV spreads were known by the fluorescent expression of eGFP in cells. In addition, we demonstrated that myostatin gene in the thigh muscles of C57BL/10 mice was knocked out by the rAAV9-SaCas9 system to make muscle mass increased obviously. The protein eGFP into rAAV has significant implications for our indirect analysis of the editing efficiency of SaCas9 in the genome of the target tissue and reduces the harm caused by off-target editing and prevents other tissue mutations. The rAAV system has substantial potential in improving muscle mass and preventing muscle atrophy. Springer Netherlands 2021-09-29 2021 /pmc/articles/PMC8510912/ /pubmed/34590222 http://dx.doi.org/10.1007/s10529-021-03183-1 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis 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/) . |
spellingShingle | Original Research Paper Weng, Shaoting Zhao, Yitian Yu, Changhong Wang, Xiaofan Xiao, Xuehan Han, Liqiang Zhang, Kunpeng Wang, Jiang Yang, Guoyu Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass |
title | Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass |
title_full | Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass |
title_fullStr | Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass |
title_full_unstemmed | Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass |
title_short | Construction of a rAAV-SaCas9 system expressing eGFP and its application to improve muscle mass |
title_sort | construction of a raav-sacas9 system expressing egfp and its application to improve muscle mass |
topic | Original Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8510912/ https://www.ncbi.nlm.nih.gov/pubmed/34590222 http://dx.doi.org/10.1007/s10529-021-03183-1 |
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