CRISPR/Cas9-mediated NlInR2 mutants:Analyses of residual mRNA and truncated proteins  

作　　者：Jun Lü Jingxiang Chen Yutao Hu Lin Chen Shihui Li Yibing Zhang Wenqing Zhang 

机构地区：[1]State Key Laboratory of Biocontrol/School of Life Sciences,Sun Yat-sen University,Guangzhou 510275,China [2]Development of Food Science and Engineering,Moutai Institute,Renhuai 564507,China

出　　处：《Journal of Integrative Agriculture》2024年第6期2006-2017,共12页农业科学学报（英文版）

基　　金：the National Natural Science Foundation of China(31730073).

摘　　要：CRISPR/Cas9 technology is a powerful genome manipulation tool in insects.However,little is known about whether mRNA and protein of a target gene are completely cleared in homozygous mutants.This study generated homozygous mutants of the insulin receptor gene 2(NlInR2)in the brown planthopper(Nilaparvata lugens)using CRISPR/Cas9 genome editing.Both frameshift mutants,E5_D17 and E6_I7,differentiated towards long wings,but there were differences in wing morphology,with E5_D17 showing wing deformities.Subsequent investigations revealed the presence of residual expression of NlInR2 mRNA in both mutants,as well as the occurrence of spliceosomes featuring exon skipping splicing in E5_D17.Additionally,the E5_D17 exhibited the detection of N-terminally truncated NlInR2 protein.RNA interference experiments indicated that the knockdown of NlInR2 expression in the E5_D17 mutant line increased the proportion of wing deformities from 11.1 to 65.6%,suggesting that the residual NlInR2 mRNA of the E5_D17 mutant might have retained some genetic functions.Our results imply that systematic characterization of residual protein expression or function in CRISPR/Cas9-generated mutant lines is necessary for phenotypic interpretation.

关 键 词：CRISPR/Cas9 Nilaparvata lugens residual mRNA skipping exon truncated protein 

分 类 号：S433[农业科学—农业昆虫与害虫防治]