The E3 ligase TRIM56 is a host restriction factor of Zika virus and depends on its RNA-binding activity but not miRNA regulation, for antiviral function

Yang, Darong; Li, Nan L.; Wei, Dahai; Liu, Baoming; Guo, Fang; Elbahesh, Husni; Zhang, Yunzhi; Zhou, Zhi; Chen, Guo-Yun; Li, Kui

HeLa cells with stable, shRNA-mediated knockdown of TRIM56 (HeLa-shT56) and control cells stably transduced with a non-targeting shRNA (HeLa-shCtrl) were described in a previous study [24] . To deplete TRIM56 expression in SVGA cells, we transduced the cells with lentiviral shRNA-094 specifically targeting human TRIM56 [24] and selected cell populations resistant to puromycin. For comparison, control cells were generated by transduction with a non-targeting control lentiviral shRNA packaged from pLKO.1-shRNA-scramble (Addgene# 1864), followed by selection in puromycin-containing medium. For transient knockdown of TLR3, a synthetic siRNA specifically targeting human TLR3 (siTLR3) [40] was transfected into cells by Lipofectamine 2000 for the indicated times as per manufacturer's instructions (Invitrogen). As a control for comparison, a non-targeting negative control siRNA (Invitrogen AM4636) was used in lieu of siTLR3.