Directing construction of CRISPR/Cas9 vector of SmPAL1 in Salvia miltiorrhiza by target efficiency detection in vitro / 中国中药杂志
Zhongguo Zhong Yao Za Zhi
; (24): 4226-4230, 2018.
Article
en Zh
| WPRIM
| ID: wpr-775354
Biblioteca responsable:
WPRO
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
Base de datos:
WPRIM
Asunto principal:
Química
/
Salvia miltiorrhiza
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Redes y Vías Metabólicas
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Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas
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Sistemas CRISPR-Cas
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Genética
Tipo de estudio:
Diagnostic_studies
Idioma:
Zh
Revista:
Zhongguo Zhong Yao Za Zhi
Año:
2018
Tipo del documento:
Article