5′ RACE-seq of HeLa cells treated with siRNA control, siZAP or siXRN1 cells and infected with wild type HIV-1 or CpG-recoded HIV-1
5′ RACE-seq of HeLa cells treated with siRNA control, siZAP or siXRN1 cells and 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 Direct-zol RNA Miniprep Plus Kit The total RNA was purified with the Monarch Spin RNA Cleanup Kit (NEB). The 5' RACE was performed from 500 ng purified RNA using the SMARTer RACE 5'/3' Kit and according to the manufacturer's instructions. 5' RACE sequencing librairies were prepared using the Native Barcoding Kit 24 V14 (Oxford Nanopore Technologies).