Wolbachia effect on small RNA response of Drosophila cells upon viral infection. Semiliki Forest Virus-Wolbachia interaction

The aim of the study is to investigate the small RNA response upon viral infection (virus-derived short RNAs and host microRNAs) in the presence/absence of Wolbachia, an bacterial symbiont that protects against RNA viruses. We used the Drosophila cells line Jw18 as a host. This cell line is infected with Wolbachia and we created a Wolbachia-free counterpart using tetracycline treatment. Both Wolbachia-infected and Wolbachia-free cells were infected or not with Semiliki Forest Virus, generating 4 different treatments: with or without Wolbachia and with or without virus. Five independent replicates were prepared for each treatment and total RNA was extracted. Small RNA libraries were then prepared using the Truseq Small RNA Library Preparation kit with an additional step to remove the Drosophila ribosomal RNA 2S before the reverse-transcription. Small RNA libraries were sequenced on one lane of HiSeq2000 instrument.