In a recent study, researchers from Zhejiang Sci-Tech University systematically identified and analyzed the BmAGO2-associated TEs, which exceed 100 in the B. morigenome. Additionally, they also mapped the small RNAs associated with BmAGO2 in B.mori using small RNA sequencing. The transposon Bm1645 is the most abundant TE associated with BmAGO2, and Bm1645-derived small RNAs represent a small RNA pool.
They determined the expression patterns of several Bm1645-derived small RNAs by northern blotting, and the results showed there was differential expression of multiple small RNAs in normal and BmNPV-infected BmN cells and silkworms from various developmental stages. They confirmed that four TE-siRNAs could bind to BmAGO2 using EMSA and also validated the recognition sites of these four TE-siRNAs in Bm1645 by dual-luciferase reporter assays. Furthermore, qRT-PCR analysis revealed the overexpression of the four TE-siRNAs could downregulate the expression of Bm1645 in BmN cells, and the transcription of Bm1645 was upregulated by the downregulation of BmAGO2.
These results suggest Bm1645 functions as a source of small RNAs and this pool can produce many BmAGO2-associated small RNAs that regulate TE’s expression.