首页|CRISPR/Cas9-mediated NIInR2 mutants:Analyses of residual mRNA and truncated proteins
CRISPR/Cas9-mediated NIInR2 mutants:Analyses of residual mRNA and truncated proteins
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国家科技期刊平台
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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(NIInR2)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 NIInR2 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 NIInR2 protein.RNA interference experiments indicated that the knockdown of NIInR2 expression in the E5_D17 mutant line increased the proportion of wing deformities from 11.1 to 65.6％,suggesting that the residual NIInR2 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.
CRISPR/Cas9Nilaparvata lugensresidual mRNAskipping exontruncated protein
Jun Lü、Jingxiang Chen、Yutao Hu、Lin Chen、Shihui Li、Yibing Zhang、Wenqing Zhang
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State Key Laboratory of Biocontrol/School of Life Sciences,Sun Yat-sen University,Guangzhou 510275,China
Development of Food Science and Engineering,Moutai Institute,Renhuai 564507,China
国家科技期刊平台
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