Egg tanning improves the efficiency of CRISPR/Cas9-mediated mutant locust production by enhancing defense ability after microinjection  

作　　者：ZHANG Ting-ting WEN Ting-mei YUE Yang YAN Qiang DU Er-xia FAN San-hong Siegfried ROTH LI Sheng ZHANG Jian-zhen ZHANG Xue-yao ZHANG Min 

机构地区：[1]Research Institute of Applied Biology,Shanxi University,Taiyuan 030006,P.R.China [2]School of Life Science,Shanxi University,Taiyuan 030006,P.R.China [3]State Key Laboratory of Cotton Biology/Key Laboratory of Plant Stress Biology,School of Life Sciences,Henan University,Kaifeng 475004,P.R.China [4]Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology,Institute of Insect Science and Technology,School of Life Sciences,South China Normal University,Guangzhou 510631,P.R.China [5]Institute for Developmental Biology,University of Cologne,Cologne 50674,Germany

出　　处：《Journal of Integrative Agriculture》2021年第10期2716-2726,共11页农业科学学报（英文版）

基　　金：This work was supported by the National Natural Science Foundation of China(32070502,31730074,32072419 and 31501905).

摘　　要：The mutant efficiency and hatching ratio are two key factors that significantly affect the construction of genome-modified mutant insects.In the construction of CRISPR/Cas9-mediated dsLmRNase2^(–/–)mutant locusts,we found that the tanned eggs which experienced a 20-min contact with the oocyst exhibited a higher success rate compared to fresh newly-laid eggs that were less tanned.However,the heritable efficiency of the dsLmRNase2 deletion to the next generation G_(1 )progeny was similar between adults derived from the tanned or less tanned engineered eggs.Further,the similar effective mutant ratios in the normally developed eggs and G_(0) adults of tanned and less tanned eggs also indicated that tanning did not reduce the absolute mutation efficiency induced by CRISPR/Cas9.Moreover,we found that the syncytial division period,which was longer than the time for tanning,conferred a window period for microinjection treatment with efficient mutation in both tanned and less tanned eggs.We further found that tanned eggs exhibited a higher hatching rate due to a reduced infection rate following microinjection.Both the anti-pressure and ultrastructure analyses indicated that the tanned eggs contained compressed eggshells to withstand increased external pressure.In summary,tanned eggs possess stronger defense responses and higher efficiency of genome editing,providing an improved model for developing Cas9-mediated gene editing procedures in locusts.

关 键 词：brown tanned eggs CRISPR/Cas9 mutant locusts MICROINJECTION defense ability 

分 类 号：S433.2[农业科学—农业昆虫与害虫防治]