strand specific RNA sequence of BmAgo2 or Siwi depleted BmN4 cells. Bombyx mori strain:BmN4

To determine which small RNA pathway mainly contributes to silencing of BmMLV RNA during persistent infection, we utilized the CRISPR/Cas9 system to generate Ago2 or Siwi knockout (KO) BmN-4 cells. We confirmed that 58% and 50% of the alleles contained the loss-of-function mutations in Ago2 and Siwi KO cells, respectively. Strand-specific RNA-seq revealed that BmMLV RNA-derived reads had a very strong sense-strand bias, and few were mapped to the antisense strand.

为明确持续性感染期间介导BmMLV RNA沉默的主要小分子RNA通路（small RNA pathway），我们采用CRISPR/Cas9系统构建了Ago2或Siwi敲除（Knockout，KO）的BmN-4细胞。我们证实，在Ago2敲除与Siwi敲除细胞中，分别有58%和50%的等位基因携带功能丧失型突变。链特异性RNA测序（Strand-specific RNA-seq）结果显示，BmMLV RNA来源的测序读段（reads）呈现极强的正义链偏好性，仅有极少部分可比对至反义链。