Directing construction of CRISPR/Cas9 vector of SmPAL1 in Salvia miltiorrhiza by target efficiency detection in vitro.
10.19540/j.cnki.cjcmm.20180726.007
- Author:
Jing-Ren QIU
1
;
Yue-Kai SU
1
;
Zhen-Qiao SONG
1
;
Xin-Sheng FANG
1
;
Jing-Yu LI
1
;
Jin ZHANG
1
;
Jian-Hua WANG
1
Author Information
1. College of Agronomy, Shandong Agricultural University, Taian 271018, China.
- Publication Type:Journal Article
- Keywords:
CRISPR/Cas9;
Salvia miltiorrhiza;
gene editing;
phenylalanine ammonia-lyase;
target efficienty detection
- MeSH:
CRISPR-Cas Systems;
Clustered Regularly Interspaced Short Palindromic Repeats;
Metabolic Networks and Pathways;
genetics;
Salvia miltiorrhiza;
chemistry;
genetics
- From:
China Journal of Chinese Materia Medica
2018;43(21):4226-4230
- CountryChina
- Language:Chinese
-
Abstract:
To construct CRISPR/Cas9 vectors for the editing of SmPAL1 in the phenylpropane metabolic pathway of Salvia miltiorrhiza, CIRSPR/Cas9 target sites of SmPAL1 were designed by online software. Its target efficiencies were detected in vitro by enzyme digestion and sequences with highly efficiency were constructed into CRISPR/Cas9 vectors. Three possible CRISPR target sequences (SmPAL1-g1, SmPAL1-g2, SmPAL1-g3) were designed and the enzyme digestion efficiencies were 53.3%, 76.6% and 10.0%. SmPAL1-g1 and SmPAL1-g2 were constructed into vector VK005-03 named as VK005-03-g1 and VK005-03-g2. The results of sequencing showed that the two CRISPR/Cas target sequences were all constructed into VK005-03. Here we first laid the foundation for the study of SmPAL1 and provided an effective strategy for the screening of sgRNA.
