RNA-seq of HeLa CRISPR control and ZAP, TRIM25 or KHNYN knockout cells mock infected or infected with wild type HIV-1 or CpG-recoded HIV-1
RNA-seq of HeLa CRISPR control and ZAP, TRIM25 or KHNYN knockout cells mock infected or infected with wild type HIV-1 or CpG-recoded HIV-1
Cells were infected at MOI=3. The medium was replaced 18 hours (h) post inoculation. Cell lysates and supernatants were collected at 48 hours post-infection. Supernatants were spun down at 300 ×g for five minutes and analysed by TZM-bl assay and immunoblotting. Total RNA was extracted using the RNeasy Mini Kit (Qiagen). RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina following manufacturer's instructions (NEB, Ipswich, MA, USA). Briefly, mRNAs were first enriched with Oligo(dT) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNAs were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by limited-cycle PCR.