[Directing construction of CRISPR/Cas9 vector of SmPAL1 in Salvia miltiorrhiza by target efficiency detection in vitro].
Zhongguo Zhong Yao Za Zhi
; 43(21): 4226-4230, 2018 Nov.
Article
en Zh
| MEDLINE
| ID: mdl-30583622
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.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Salvia miltiorrhiza
/
Redes y Vías Metabólicas
/
Sistemas CRISPR-Cas
Tipo de estudio:
Diagnostic_studies
Idioma:
Zh
Revista:
Zhongguo Zhong Yao Za Zhi
Año:
2018
Tipo del documento:
Article
País de afiliación:
China