Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models

In this protocol, we describe how to utilize the unnatural amino acid (UAA) incorporation system to read through endogenous premature termination codons in a Duchenne muscular dystrophy mouse model. We detail how to screen and optimize tRNA-enzyme pairs for efficient UAA incorporation, deliver the s...

Descripción completa

Detalles Bibliográficos
Autores principales: Zheng, Zhetao, Shi, Ningning, Xia, Qing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9792948/
https://www.ncbi.nlm.nih.gov/pubmed/36527714
http://dx.doi.org/10.1016/j.xpro.2022.101950
_version_ 1784859746472296448
author Zheng, Zhetao
Shi, Ningning
Xia, Qing
author_facet Zheng, Zhetao
Shi, Ningning
Xia, Qing
author_sort Zheng, Zhetao
collection PubMed
description In this protocol, we describe how to utilize the unnatural amino acid (UAA) incorporation system to read through endogenous premature termination codons in a Duchenne muscular dystrophy mouse model. We detail how to screen and optimize tRNA-enzyme pairs for efficient UAA incorporation, deliver the system intraperitoneally or intramuscularly in pathogenic mice by an adeno-associated viral (AAV) vector, and evaluate the restoration of endogenous dystrophin and increase in muscle strength after AAV injection. For complete details on the use and execution of this protocol, please refer to Shi et al. (2021).(1)
format Online
Article
Text
id pubmed-9792948
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-97929482022-12-28 Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models Zheng, Zhetao Shi, Ningning Xia, Qing STAR Protoc Protocol In this protocol, we describe how to utilize the unnatural amino acid (UAA) incorporation system to read through endogenous premature termination codons in a Duchenne muscular dystrophy mouse model. We detail how to screen and optimize tRNA-enzyme pairs for efficient UAA incorporation, deliver the system intraperitoneally or intramuscularly in pathogenic mice by an adeno-associated viral (AAV) vector, and evaluate the restoration of endogenous dystrophin and increase in muscle strength after AAV injection. For complete details on the use and execution of this protocol, please refer to Shi et al. (2021).(1) Elsevier 2022-12-16 /pmc/articles/PMC9792948/ /pubmed/36527714 http://dx.doi.org/10.1016/j.xpro.2022.101950 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Zheng, Zhetao
Shi, Ningning
Xia, Qing
Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models
title Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models
title_full Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models
title_fullStr Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models
title_full_unstemmed Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models
title_short Adeno-associated viral delivery of engineered tRNA-enzyme pairs into nonsense mutation mouse models
title_sort adeno-associated viral delivery of engineered trna-enzyme pairs into nonsense mutation mouse models
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9792948/
https://www.ncbi.nlm.nih.gov/pubmed/36527714
http://dx.doi.org/10.1016/j.xpro.2022.101950
work_keys_str_mv AT zhengzhetao adenoassociatedviraldeliveryofengineeredtrnaenzymepairsintononsensemutationmousemodels
AT shiningning adenoassociatedviraldeliveryofengineeredtrnaenzymepairsintononsensemutationmousemodels
AT xiaqing adenoassociatedviraldeliveryofengineeredtrnaenzymepairsintononsensemutationmousemodels

Ejemplares similares