RT Journal Article
SR Electronic
T1 Collateral cleavage of 28s rRNA by RfxCas13d causes death of mice
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.01.17.476700
DO 10.1101/2022.01.17.476700
A1 Yunfei Li
A1 Junjie Xu
A1 Xuefei Guo
A1 Zhiwei Li
A1 Lili Cao
A1 Shengde Liu
A1 Ying Guo
A1 Guodong Wang
A1 Yujie Luo
A1 Zeming Zhang
A1 Xuemei Wei
A1 Yingchi Zhao
A1 Tongtong Liu
A1 Xiao Wang
A1 Huawei Xia
A1 Ming Kuang
A1 Qirui Guo
A1 Junhong Li
A1 Luoying Chen
A1 Yibing Wang
A1 Qi Li
A1 Fengchao Wang
A1 Qinghua Liu
A1 Fuping You
YR 2022
UL http://biorxiv.org/content/early/2022/01/18/2022.01.17.476700.abstract
AB The CRISPR-Cas13 system is an RNA-guided RNA-targeting system, and has been widely used in transcriptome engineering with potentially important clinical applications. However, it is still controversial whether Cas13 exhibits collateral activity in mammalian cells. Here, we found that knocking down gene expression using RfxCas13d in the adult brain neurons caused death of mice, which was not resulted from the loss of target gene function or off-target effects. Mechanistically, we showed that RfxCas13d exhibited collateral activity in mammalian cells, which is positively correlated with the abundance of target RNA. The collateral activity of RfxCas13d could cleave 28s rRNA into two fragments, leading to translation attenuation and activation of the ZAKα-JNK/p38-immediate early gene (IEG) pathway. These results provide new mechanistic insights into the collateral activity of RfxCas13d and warn that the biosafety of CRISPR-Cas13 system needs further evaluation before applying it to clinical treatments.Competing Interest StatementThe authors have declared no competing interest.