[Expression of human micro-dystrophin gene after retrovirus infection in mdx mice bone marrow-derived mesenchymal stem cells].
OBJECTIVE
To construct the retroviral vector containing human micro-dystrophin gene and detect the expression of human micro-dystrophin in mdx mice bone marrow-derived mesenchymal stem cells (MSCs) after retrovirus infection.


METHODS
Retroviral vector for micro-dystrophin gene was constructed and transferred into the packing cell PA317 mediated by Lipofectamine 2000. The retroviral supernatant containing the target genes were subsequently used to infect mdx mice MSCs. Micro-dystrophin expression was examined by methods of immunofluorescence staining and reverse transcriptase-polymerase chain reaction.


RESULTS
Micro-dystrophin retroviral vector was successfully constructed and transferred into PA317 cells, and 48 h after infection with the recombinant retrovirus in mdx mice MSCs, 319 bp fragment could be detected by electrophoresis in the RT-PCR products. The red particles could be detected in some infected mdx mice MSCs with immunofluorescence staining. CONCLUSION mdx mice MSCs infected with retrovirus containing micro-dystrophin gene can express micro-dystrophin protein.
Yu
Mei-Juan
MJ
Stem Cells and Tissue Engineering Research Center, Sun Yat-Sen University, Guangzhou 510080, China.
Zhang
Cheng
C
Wang
Shu-Hui
SH
Zhang
Ya-Ni
YN
Yao
Xiao-Li
XL
Lu
Xi-Lin
XL
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D016189:Dystrophin / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D018101:Mice, Inbred mdx; D009137:Muscular Dystrophy, Animal / Q000378:metabolism; D012192:Retroviridae Infections; D014162:Transfection
D016189:Dystrophin