RNA-seq of Monkeypox virus-protein overexpression in human embryonic stem cells (hESCs) revealed the effects of H3L, A29L, A35R and I1L in human

Monkeypox virus (MPV) can infect human cells and induces injuries in human. However, whether and how MPV affect the function of human cells remain to be elucidated. Here, we overexpressed H3L, A29L, A35R and I1L in hESCs, separately. We appliced RNA-seq on these hESCs and want to know gene expression patterns of control and Overexpression hESCs. Subsequent data analyses will help us to know whether and how H3L, A29L, A35R or I1L affect cells in human. We separately cloned DNA seqeunces of H3L, A29L, A35R and I1L into lentivirus and infected hESCs, follwed with puro screening. The stable cell lines were rotinely maintained in mTesR1 medium in 6-well plate. Stable cell lines were collected for RNA purification and RNA-seq. Lenti-virus control cell line (control hESCs) did not express any protein. Total RNAs were extracted and purified with RNeasy Mini Kit (Qiagen). 4 biological replicates were applied for each overexpression cell line.

猴痘病毒（Monkeypox virus, MPV）可感染人类细胞并对人体造成损伤。然而，该病毒是否以及如何影响人类细胞的功能仍有待阐明。本研究中，我们分别在人类胚胎干细胞（human embryonic stem cells, hESCs）中过表达H3L、A29L、A35R与I1L基因，并对这些hESCs进行RNA测序（RNA-seq），以探究对照组与过表达组hESCs的基因表达谱。后续数据分析将有助于阐明H3L、A29L、A35R或I1L对人类细胞的影响及其作用机制。我们分别将H3L、A29L、A35R及I1L的DNA序列克隆至慢病毒载体，随后感染hESCs并进行嘌呤霉素筛选。稳定细胞系于6孔板中使用mTesR1培养基常规培养，收集后用于RNA纯化及RNA测序。慢病毒对照组细胞系（对照hESCs）不表达任何外源蛋白。总RNA采用RNeasy Mini试剂盒（Qiagen）进行提取与纯化。每个过表达细胞系均设置4个生物学重复。