Construction of a lentiviral low expression of miR-139-5 p vector and validation of its transduction efficiency in H9 c2 cells
10.3969/j.issn.1001-1978.2015.01.027
- VernacularTitle:miR-139-5p低表达重组慢病毒的构建及其在H9c2细胞的表达验证
- Author:
Yuan JIANG
;
Zhuojun HUANG
;
Zhaoyan QIANG
;
Yanna WU
- Publication Type:Journal Article
- Keywords:
miR-139-5 p;
lentiviral vectors;
H9 c2 cells;
infection;
qRT-PCR;
low expression
- From:
Chinese Pharmacological Bulletin
2015;(1):127-130
- CountryChina
- Language:Chinese
-
Abstract:
Aim To construct a lentiviral low expres-sion of miR-139-5 p vector and validate its expression efficiency in H9c2 cells. Method Target sequence was designed according to the sequence of rat miR-139-5p. Oligonucleotide duplex was synthesized and cloned into the lentiviral vector pGC-LV. The recombi-nant lentiviral vector, pHelper 1. 0, and pHelper 2. 0 were co-transfected into 293T cells, packaging virus. Then H9 c2 cells were infected with the supernatant containing lentiviral particles, and its infection effi-ciency and miR-139-5 p expression were determined by fluorescent microscope and real-time quantitative PCR, respectively. Results A lentiviral low expression of miR-139-5p vector was successfully constructed. The infection efficiency in H9c2 cells reached over 95%, and the relative expression of miR-139-5p was significantly down-regulated. Conclusion The lentiviral low expression of miR-139-5p vector is successfully constructed, and the expression of miR-139-5p in infected H9c2 cells is inhibited effectively.
