Construction and identification of Kir2ds4 RNAi lentiviral vector.
- Author:
Li-Ping DOU
1
;
Wan-Ming DA
;
Chang WANG
;
Hui-Yuan KANG
;
Yu ZHAO
;
Jing-Fen SUN
;
Hai-Jie JIN
;
Quan-Shun WANG
;
Chun-Ji GAO
;
Li YU
Author Information
1. Department of Hematology, General Hospital of PLA, Beijing 100853, China.
- Publication Type:Journal Article
- MeSH:
Base Sequence;
Genetic Vectors;
genetics;
Green Fluorescent Proteins;
genetics;
metabolism;
Humans;
Lentivirus;
genetics;
metabolism;
Molecular Sequence Data;
RNA Interference;
RNA, Small Interfering;
genetics;
metabolism;
Receptors, KIR;
biosynthesis;
genetics
- From:
Journal of Experimental Hematology
2008;16(3):663-666
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
