Abstract
PIWI-interacting RNAs (piRNAs) and their associated PIWI proteins are essential for germ line development and have been found in Bombyx mori. In the past decade genomic editing tools have been developed, one of which, called CRISPR has arisen as an attractive option due to its ease of use and cost efficient methods. This study will focus on adopting the CRISPR system in the Bombyx mori cell culture line BmN4s in the first step to using CRISPR to understand piRNA biogenesis in Bombyx mori. We demonstrate that sgRNAs in conjunction with Cas9, targeting ZsGreen can reduce but not eliminate the levels of ZsGreen fluorescence. However, targeting proteins involved in piRNA biogenesis using both normal and modified sgRNA designs such as Siwi has yet to be observed via protein and genetic analysis. Future studies will focus on optimization of the CRISPR/Cas9 system before studying factors involved in the piRNA pathway in Bombyx mori.