Construction and characterization of RNAi lentiviral vector targeting human CD106 gene
10.3969/j.issn.1001-3733.2016.06.008
- VernacularTitle:靶向人CD106基因RNAi慢病毒载体的构建与鉴定
- Author:
Legang SUN
;
Tingting YU
;
Ling LIU
;
Zhaohui LI
;
Honghai FU
;
Lifang WANG
- Keywords:
CD106;
RNAi;
Lentiviral vector
- From:
Journal of Practical Stomatology
2016;32(6):783-786
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct CD106-targeted RNAi lentiviral vector plasmids. Methods:4 targets aimed at CD106(Target 1, 2, 3, 4)were designed. Oligo-DNA fragment containing short hairpin frame was synthesized and reannealed, and then cloned into lentivi-ral expression vector. PCR and sequencing analysis were made for verifying the positive clones. The virus packaging plasmids were trans-fected into 293T cells to harvest siRNA lentivirus. After infection in HN12 cells, Real-time PCR and western blot were performed to de-termine the expressing level of CD106. Results:PCR and sequencing revealed that siRNA plasmids was correctly constructed. Virus with a titer of 1 × 109 TU/ml was successfully packaged at least. CD106 expression in HN12 cells could be knockdown by virus infection sig-nifically, compared with negative control lentivirus. Conclusion: The recombinant lentiviral siRNA expressing vector targeting human CD106 gene has been successfully constructed and packaged. CD106 gene in cells may be down-regulated by lentiviral siRNA.
