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Published on: October 7, 2018
[Construction and identification of a recombinant lentivirus harboring small interfering RNA targeting mouse CD99
Gong Zhang1, Fang Liu, Xiao-yan Chen
1Department of Pathology, Nanfang Hospital, Southern Medical University, and Key Lab for Molecular Tumor Pathology of Guangdong Province, Southern Medical University, Guangzhou 510515, China. zhanggong7446@163.com
Objective:
To construct a recombinant lentivirus harboring RNA interference sequence targeting mouse CD99 antigen-like 2 (mCD99L2) gene and observe its infection efficiency of 293FT cells.
Methods:
Four pairs of small interfering RNAs (siRNAs) targeting mCD99L2 cDNA were designed, synthesized and linked to the lentivirus vector SD1259 to construct the lentivirus shuttle plasmids. After sequencing, the 4 lentivirus shuttle plasmids were transfected into 293FT cells in the presence of packaging plasmids. Forty-eight hours later, the supernatant was collected and the titer and infection efficiency of the recombinant lentivirus were determined according to the expression of the reporter gene enhanced green fluorescent protein (EGFP) under fluorescent microscope.
Results:
DNA sequencing demonstrated that mCD99L2 siRNAs were successfully cloned to the lentiviral vector SD1259. The titer of concentrated virus was 1x10(7)/ml in the supernatant of the infected cells.
Conclusion:
The recombinant lentivirus containing siRNA targeting mCD99L2 gene has been successfully constructed, which provide the basis for future establishment of visualized cell model and animal model of Hodgkin's lymphoma.
