Genome-wide CRISPR-Cas screen identifies CRNKL-1 as a nuclear retention factor of HIV-1 unspliced RNA

ABSTRACT: The HIV-1 regulatory Rev protein binds to unspliced or incompletely-spliced HIV-1 RNAs and thereby targets these RNAs for nuclear export and translation. Without Rev, these intron-retaining RNAs are trapped in the nucleus. To identify cellular factors involved in nuclear trapping of intron-retaining HIV-1 RNAs, a genome-wide lentiviral CRISPR/Cas knockout vector library was screened for enhanced reporter gene expression from the unspliced HIV-1 RNA. Reporter cell line establishment A dual-color proviral reporter plasmid, HIV-dual-GT, was first constructed. gag is ligated by a BFP; nef is replaced by a DsRed; rev, pol and other accerssory genes are mutatd by point mutations. tat remains functional for active transcription. Jurkat CD4 T cells were transduced by this proviral vector. Two c

SUBMITTER: Han Xiao 

PROVIDER: S-BSST243 | biostudies-other |

REPOSITORIES: biostudies-other