[Regulation of gene co-expression after bicistronic retrovirus-mediated gene transfer into human cord blood CD34+ cells].
OBJECTIVE
To compare internal SV40 early promoter with IRES of the in regulatory effect on gene co-expression after retrovirus-mediated gene transfer into human cord blood CD34+ cells.


METHODS
Based on pLXSN, vectors pLESN and pLEIN carrying EGFP and neo gene, distinguished by the in sequences between two, genes, were constructed. After liposome-mediated gene transfer into packaging cells, high-titre virus supernatant was obtained. After supernatant infection in vitro, transduced cells were cultured in CFC assay.


RESULTS
FACS analysis demonstrated that 10% of human cord blood CD34+ cell were infected in vitro. All of G418 resistant colonies demonstrated green fluorescence in LEIN group, while only 41.1% in LESN group.


CONCLUSIONS
IRES is superior to SV40 promoter for the guarantee of co-expression in bicistronic retrovirus-mediated gene therapy in human cord blood CD34+ cells.
Liu
B
B
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850.
Wu
Y
Y
Tong
Y
Y
D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis*; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D051379:Mice; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics
D018952:Antigens, CD34