Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine

N(6)-methyladenosine (m(6)A) modification of human immunodeficiency virus type 1 (HIV-1) RNA plays a critical role in regulating viral replication and evasion of innate immunity. Here, we describe a protocol for the production of HIV-1 with altered m(6)A levels by manipulating the expression of m(6)...

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Detalles Bibliográficos
Autores principales: Phillips, Stacia, Baek, Alice, Kim, Sanggu, Chen, Shuliang, Wu, Li
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9389413/
https://www.ncbi.nlm.nih.gov/pubmed/35990737
http://dx.doi.org/10.1016/j.xpro.2022.101616
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author Phillips, Stacia
Baek, Alice
Kim, Sanggu
Chen, Shuliang
Wu, Li
author_facet Phillips, Stacia
Baek, Alice
Kim, Sanggu
Chen, Shuliang
Wu, Li
author_sort Phillips, Stacia
collection PubMed
description N(6)-methyladenosine (m(6)A) modification of human immunodeficiency virus type 1 (HIV-1) RNA plays a critical role in regulating viral replication and evasion of innate immunity. Here, we describe a protocol for the production of HIV-1 with altered m(6)A levels by manipulating the expression of m(6)A demethylases in HIV-1 producer cells. RNA from purified virions is analyzed by northern blot and dot blot for m(6)A levels prior to use in downstream assays to determine the function of m(6)A modification of viral RNA. For complete details on the use and execution of this protocol, please refer to Chen et al. (2021).
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spelling pubmed-93894132022-08-20 Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine Phillips, Stacia Baek, Alice Kim, Sanggu Chen, Shuliang Wu, Li STAR Protoc Protocol N(6)-methyladenosine (m(6)A) modification of human immunodeficiency virus type 1 (HIV-1) RNA plays a critical role in regulating viral replication and evasion of innate immunity. Here, we describe a protocol for the production of HIV-1 with altered m(6)A levels by manipulating the expression of m(6)A demethylases in HIV-1 producer cells. RNA from purified virions is analyzed by northern blot and dot blot for m(6)A levels prior to use in downstream assays to determine the function of m(6)A modification of viral RNA. For complete details on the use and execution of this protocol, please refer to Chen et al. (2021). Elsevier 2022-08-10 /pmc/articles/PMC9389413/ /pubmed/35990737 http://dx.doi.org/10.1016/j.xpro.2022.101616 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
Phillips, Stacia
Baek, Alice
Kim, Sanggu
Chen, Shuliang
Wu, Li
Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine
title Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine
title_full Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine
title_fullStr Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine
title_full_unstemmed Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine
title_short Protocol for the generation of HIV-1 genomic RNA with altered levels of N(6)-methyladenosine
title_sort protocol for the generation of hiv-1 genomic rna with altered levels of n(6)-methyladenosine
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9389413/
https://www.ncbi.nlm.nih.gov/pubmed/35990737
http://dx.doi.org/10.1016/j.xpro.2022.101616
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