[Construction and identification of a recombinant lentivirus harboring small interfering RNA targeting mouse CD99 antigen-like 2 gene].

Abstract

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.

• Authors: Gong Zhang, Fang Liu, Xiao-yan Chen, Wei-yi Fang, Hui-ling Li, Zuo-ping Huang, Hong-juan Chu, Xin Wang, Tong Zhao
• Journal: Nan fang yi ke da xue xue bao = Journal of Southern Medical University (Nan Fang Yi Ke Da Xue Xue Bao) Vol. 29 Issue 2 Pg. 228-31 (Feb 2009) ISSN: 1673-4254 [Print] China
• PMID: 19246285 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 12E7 Antigen
• Antigens, CD
• Cd99 protein, mouse
• DNA, Complementary
• RNA, Small Interfering
• Recombinant Proteins
• enhanced green fluorescent protein
• Green Fluorescent Proteins

Topics

• 12E7 Antigen
• Animals
• Antigens, CD (genetics)
• Base Sequence
• DNA, Complementary (genetics)
• Genetic Vectors
• Green Fluorescent Proteins (genetics, metabolism)
• Hodgkin Disease (pathology)
• Lentivirus (genetics, metabolism)
• Mice
• Molecular Sequence Data
• RNA Interference
• RNA, Small Interfering (genetics)
• Recombinant Proteins (genetics)
• Tumor Cells, Cultured