[Retrovirus vector expresses luciferase and TPO gene under the control of doxycycline in NIH3T3 cells].
OBJECTIVE
To use the retrovirus vector Tet-On regulation system to control the expression of TPO and luciferase gene by doxycycline (Dox).


METHODS
Packing the recombinant Tet-On retrovirus, and infecting it to NIH3T3 cells to establish recombined Tet-On retrovirus cell lines, RevTet-On3T3. To construct the pRevTRETPO, the new cell line was transferred to PT67 cell to package the recombinant retrovirus which contained both the TRE and TPO. This virus was RevTet-On3T3 used to establish the two inserting recombinant retrovirus cell strain RevTet-On3T3TPO. Doxycycline was used to control the expression of TPO in RevTet-On3T3.


RESULTS
When Dox (5 mg/L) was added to the RevTet-On3T3 cells, a higher activity of luciferase was seen (mean, 5.9 x 10(4) RLU.S) compared with the mean activity (2.6 x 10(3) RLU.S) in the absence of Dox. When Dox was added to the RevTet-On3T3TPO cells, cell populations had high TPO expression (mean, 12.8 ng/ml) in 5 mg/L of Dox (On state), and a low TPO expression (mean, 0.56 ng/ml) in the absence of Dox (Off state).


CONCLUSIONS
The RevTet-On gene expression system could modulate the expression of multiple genes by tetracycline and its derivatives, providing a useful tool for gene therapy.
Wei
X
X
Institute of Hematology, People's Hospital, Beijing Medical University, Beijing 100044, China.
Fu
S
S
Wu
S
S
D016475:3T3 Cells; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004318:Doxycycline / Q000494:pharmacology*; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D005822:Genetic Vectors; D008156:Luciferases / Q000235:genetics*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D013926:Thrombopoietin / Q000235:genetics*
D000900:Anti-Bacterial Agents; D013926:Thrombopoietin; D008156:Luciferases; D004318:Doxycycline