| Title: | Functional characterization of SlitPBP3 in Spodoptera litura by CRISPR/Cas9 mediated genome editing |
| Author(s): | Zhu GH; Xu J; Cui Z; Dong XT; Ye ZF; Niu DJ; Huang YP; Dong SL; |
| Address: | "Education Ministry Key Laboratory of Integrated Management of Crop Disease and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai Institutes for Biological Sciences, Shanghai 200032, China. Education Ministry Key Laboratory of Integrated Management of Crop Disease and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: sldong@njau.edu.cn" |
| DOI: | 10.1016/j.ibmb.2016.05.006 |
| ISSN/ISBN: | 1879-0240 (Electronic) 0965-1748 (Linking) |
| Abstract: | "Functional gene analysis by using genome editing techniques is limited only in few model insects. Here, we reported an efficient and heritable gene mutagenesis analysis in an important lepidopteran pest, Spodoptera litura, using the CRISPR/Cas9 system. By using this system, we successfully obtained the homozygous S. litura strain by targeting the pheromone binding protein 3 gene (SlitPBP3), which allowed us to elucidate the role of this gene in the olfaction of the female sex pheromones. By co-injection of Cas9 mRNA and sgRNA into S. litura eggs, highly efficient chimera mutation in SlitPBP3 loci was detected both in injected eggs (39.1%) and in the resulting individual moths (87.5%). We used the mutant moths as parents to obtain the G1 offspring and the homozygous mutant strain in G2. The function of SlitPBP3 was explored by Electroantennogram (EAG) recordings with a homozygous mutant strain. The result showed that the EAG responses were significantly decreased in mutant males than in control males when treated with the major sex pheromone component (Z9,E11-14:Ac) and a minor component (Z9-14:Ac) at higher dosages. The results demonstrate that s SlitPBP3 gene plays a minor role in the perception of the female sex pheromones. Furthermore, our study provides a useful methodology with the CRISPR/Cas9 system for gene in vivo functional study, particular for lepidopteran species in which the RNAi approach is not efficient" |
| Keywords: | "Animals Base Sequence Carrier Proteins/chemistry/*genetics/metabolism *Chemotaxis Cloning, Molecular *Clustered Regularly Interspaced Short Palindromic Repeats DNA, Complementary/genetics/metabolism Female Gene Editing Insect Proteins/chemistry/*genetics/;" |
| Notes: | "MedlineZhu, Guan-Heng Xu, Jun Cui, Zhen Dong, Xiao-Tong Ye, Zhan-Feng Niu, Dong-Juan Huang, Yong-Ping Dong, Shuang-Lin eng Research Support, Non-U.S. Gov't England 2016/05/19 Insect Biochem Mol Biol. 2016 Aug; 75:1-9. doi: 10.1016/j.ibmb.2016.05.006. Epub 2016 May 15" |
