Year: 2022
Type: article
Abstract: 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) .
Source: STAR Protocols
Institutions: University of Iowa, The Ohio State University, Wuhan University
Language: English
Topic: RNA modifications and cancer
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,500
Awards: G1258833559, G1412571906, G1573650897, Mechanisms of HIV-1 RNA Methylation in Regulating Viral Replication, Targeting HIV-1 RNA modifications in latently infected CD4+ T cells for therapeutic development, G3363898203, SAMHD1-mediated regulation of HIV-1 innate immunity and viral gene expression, HIV-1-induced upregulation of m6A modifications of cellular RNA in CD4+ T-cells, G5858236458, G630064964, G8371663866, G9000168083, G9000177849, G9000213050, G9000221277, G9000265723, G9000290112, Cancer Center Support Grant, Targeting HIV-1 RNA modifications in latently infected CD4+ T cells for therapeutic development
OpenAlex ID: W4293152486