Small RNA-seq analysis of human airway epithelium infected by Human Bocavirus 1

Human Bocavirus 1(HBoV1), which belongs to the genus Bocaparvovirus of the family Parvoviridae, infects well differentiated human airway epithelium which is at mitotically quiescent state. To systematicaly investigate the host and viral small RNA expression after HBoV1 infection of primary human airway epithelium cultured at an air-liquid interface (HAE-ALI), small RNA-seq was applied to study the small RNA transcriptome profile of HAE-ALI infected by HBoV1. Primary human airway epithelial cells were cultured on collagen-coated, semipermeable membrane inserts of 0.33 cm2 and were polarized/differentiated at an ALI for 3-4 weeks. The well differentiated HAE-ALI cells were infected with HBoV1 at a multiplicity of infection (MOI) of 1,000 vgc/cell. At 7 days post infection, HBoV1 infected-cell were collected and RNA samples were prepared using the miRNeasy Mini Kit by following the manufacturer’s instruction. HAE cells from two Transwells were used as one RNA sample, totally 6 RNA samples from HBoV1 infected HAE-ALI cultures were performed for RNA isolation. All RNA samples were analyzed on an Agilent 2100 Bioanalyzer with an Agilent RNA 6000 Nano Kit to ensure an RNA Integrity Number (RIN) ≥ 8.0. Three RNA samples were chosen randomly for small RNA-seq.