GSM6284790: A1_RNA; Homo sapiens; RNA-Seq
ZIKV IbH 30656 NR-50066 was isolated from the blood of a human in Ibadan, Nigeria and was obtained by M.K. through BEI Resources, NIAID, NIH, as part of the WRCEVA program. DENV-2 strain 16681 was obtained by M.K. from the World Arbovirus Reference Center, University of Texas Medical Branch, Galveston, TX. 6-well plates were coated with poly-L-Ornithine (PLO) and laminin before seeding hNPCs ( 8x105 cells per well) (Topol et al., 2015). 24h post-plating the cells were infected with ZIKV or DENV at a multiplicity of infection of 1 and incubated at 37°C. Viral inocula were removed at 3 hours post infection (hpi). Cells were collected and processed for ribosome footprinting at 72 hpi. Human neuronal progenitor cells (hNPCs) (n=2) were infected with ZIKV-Ibh and DV-2 16681 (72 hr) followed by cycloheximide treatment for 10 minutes. Total RNA and ribosome-protected fragments were isolated following published protocol (Ingolia et al., 2012). Small RNA libraries were generated using SMARTer smRNA kit from Illumina. Deep sequencing libraries were generated from these fragments and sequenced on the HiSeq 2500 platform. Genome annotation was from the human genome sequence GRCh37 downloaded from Ensembl public database: http:// www.ensembl.org. For virus genome we used the reference genomes for ZIKV-Ibh and DV-2 16681 downloaded from the NCBI database (Coordinators, 2016).