Dual inducible vectors and cell lines
View Patent ↗The invention pertains to a novel cell line, an HIV tat-rev dependent GFP-Gaussia luciferase Reporter cell line, known henceforth as the GGR cell line, that detects pseudotype and replication competent HIV (cloned or uncloned isolates, in cell media or human serum) rapidly and with high sensitivity. This GGR cell line provides an improved method of characterizing the entry phenotype of HIV envelope genes, and detecting and examining primary HIV samples in the context of laboratory research, clinical trial monitoring, and medical diagnostics. Examples include, but are not limited to, determining the functional HIV viral load, responsiveness to treatment, characterization of viral co-receptor usage (testing for viral co-receptor usage, i.e., CCR5 vs CXCR4, as required prior to prescribing FDA-approved CCR5 inhibitors), and characterization of other viral or drug resistance phenotypic properties to guide treatment.
1. A vector comprising the transactivators of the tetracycline and ponasterone inducible systems and the FRT-LacZeo selection cassette wherein the vector allows inducible expression of two genes of interest and a high expression of the selectable markers LacZ and Zeocin, wherein the transactivator of tetracycline is operably linked to VGECR of the transactivator of ponasterone by a P2A ribosome skipping sequence, and wherein a RSV 5′ LTR promoter is operably linked to RKR of the transactivator of ponasterone.
2. The vector according to claim 1 , wherein the vector is represented in FIG. 13 .
3. A cell line comprising the vector identified in claim 1 .
4. A method for creating a cell line that can be made to regulate the expression of any two genes placed under the respective inducible promoter comprising transfecting a cell with the vector of FIG. 13 .
5. A vector as set forth in FIG. 32 .
6. A cell line comprising the vector identified in claim 5 .
7. A cell line created from the method according to claim 4 .
8. A method for controlling the expression of any two genes of interest comprising transfecting a cell with the vector of FIG. 13 .