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


CONCLUSIONS
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.
Zhang
Gong
G
Department 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
Liu
Fang
F
Chen
Xiao-yan
XY
Fang
Wei-yi
WY
Li
Hui-ling
HL
Huang
Zuo-ping
ZP
Chu
Hong-juan
HJ
Wang
Xin
X
Zhao
Tong
T
D000071443:12E7 Antigen; D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006689:Hodgkin Disease / Q000473:pathology*; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D014407:Tumor Cells, Cultured
D000071443:12E7 Antigen; D015703:Antigens, CD; C522605:Cd99 protein, mouse; D018076:DNA, Complementary; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins