The clustered regularly interspaced short palindromic repeats/associated proteins system for the induction of gene mutations and phenotypic changes in Bombyx mori.
To probe the general phenomena of gene mutations, Bombyx mori, the lepidopterous model organism, was chosen as the experimental model. To easily detect phenotypic variations, the piggyBac system was utilized to introduce two marker genes into the silkworm, and 23.4% transposition efficiency aided in easily breeding a new strain for the entire experiment. Then, the clustered regularly interspaced short palindromic repeats/an associated protein (Cas9) system was utilized. The results showed that the Cas9 system can induce efficient gene mutations and the base changes could be detected since the G0individuals in B. mori; and that the mutation rates on different target sites were diverse. Next, the gRNA2-targeted site that generated higher mutation rate was chosen, and the experimental results were enumerated. First, the mutation proportion in G1generation was 30.1%, and some gene mutations were not inherited from the G0generation; second, occasionally, base substitutions did not lead to variation in the amino-acid sequence, which decreased the efficiency of phenotypic changes compared with that of genotypic changes. These results laid the foundation for better use of the Cas9 system in silkworm gene editing.
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Jia
J
Song
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Jiaqian
J
Che
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Zhengying
Z
You
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Xiaogang
X
Ye
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Jisheng
J
Li
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Lupeng
L
Ye
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Yuyu
Y
Zhang
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Qiujie
Q
Qian
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China bxzhong@zju.edu.cn.
Boxiong
B
Zhong
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics*; D064113:CRISPR-Cas Systems; D019476:Insect Proteins / Q000235:genetics*; D009154:Mutation; D010641:Phenotype
D019476:Insect Proteins
Bombyx mori; CRISPR/Cas9; gene editing; inherited mutant; phenotypic change