PRJNA542626
Influenza A virus (A/Puerto Rico/8/1934(H1N1))
5' RACE was performed to investigate cap-snatching from an IAV mini-gene system. Human lung epithelial cells (A549) cells were co-transfected with plasmids expressing the A/Puerto Rico/8/1934 (H1N1) RdRp subunits (i.e. PA, PB1, PB2, and NP) and a mini-gene construct expressing a negative-sense RNA transcript comprising the Firefly luciferase ORF flanked by various UTRs from A/Puerto Rico/8/1934 (H1N1). mRNA was extracted and host primers profiled by high-throughput sequencing using Illumina HiSeq 2000. Four A/Puerto Rico/8/1934 (H1N1) UTR combinations were tested: 5'NP-Luc-NP3' (NPhomo), 5'PB1-Luc-PB13' (PB1homo), 5'PB1-Luc-NP3' (NPhetero), and 5'NP-Luc-PB13' (PB1hetero).