Study the balance of T cell dynamics in autoimmunity, inflammation, and cancer
Simplify your studies of T cell dynamics with SBI’s ready-to-use, pre-built Treg and Th17 vectors and cell lines. With pMSCV-Mouse RORγt-EF1α-GFP-T2A-Puro for RORγt Overexpression, you can choose between three different ways to study RORγt overexpression from the MSCV promoter for strong expression in hematopoietic and stem cells:
- Lentivector-based for transfection-resistant cell lines
Create stable RORγt-overexpressing cell lines by integrating the pMSCV-Mouse RORγt-EF1α-GFP-T2A-Puro expression cassette into the genome using SBI’s highly regarded and reliable lentivector technology. With our lentivector technology, you can take advantage of our high-titer lentivector packaging system to efficiently deliver your RORγt overexpression cassette to cells using transduction—great for transfection-resistant cell lines.
- PiggyBac Transposon-based* for infection-resistant cell lines
Create stable RORγt-overexpressing cell lines by integrating the pMSCV-Mouse RORγt-EF1α-GFP-T2A-Puro expression cassette into the genome with SBI’s easy and consistent PiggyBac Transposon System. With our PiggyBac Transposon System, you get easy and consistent transgenesis for RORγt overexpression in infection-resistant cell lines.
Pre-built Jurkat cell line to save time and labor
Quickly get your Treg studies off the ground with our pre-built Jurkat cell line that already has the pMSCV-Mouse RORγt-EF1α-GFP-T2A-Puro expression cassette integrated into the genome. We transduced Jurkat cells with the pMSCV-Mouse RORγt-EF1α-GFP-T2A-Puro expression cassette and have validated RORγt overexpression, so you don’t have to—just culture cells and start your studies.
Academic customers can purchase PiggyBac Transposon System components for internal research purposes for indefinite use, whereas commercial customers must sign a customer agreement for a six-month, limited-use license to test the technology.
For end user license information, see the following:
* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Transposagen Biopharmaceuticals, Inc.
How It Works
A quick introduction to SBI’s lentivector packaging and PiggyBac Transposon technologies
Start producing lentivirus
The PiggyBac Transposon System’s Cut-and-Paste Mechanism
The efficient PiggyBac Transposon System uses a cut-and-paste mechanism to transfer DNA from the PiggyBac Vector into the genome. If only temporary genomic integration is desired, the Excision-only PiggyBac Transposase can be transiently expressed for footprint-free removal of the insert, resulting in reconstitution of the original genome sequence.
Figure 1. The PiggyBac Transposon System’s cut-and-paste mechanism.
- The Super PiggyBac Transposase binds to specific inverted terminal repeats (ITRs) in the PiggyBac Cloning and Expression Vector and excises the ITRs and intervening DNA.
- The Super PiggyBac Transposase inserts the ITR-Expression Cassette-ITR segment into the genome at TTAA sites.
- The Excision-only Super PiggyBac Transposase can be used to remove the ITR-Expression Cassette-ITR segment from the genome, for footprint-free removal
Robust overexpression of RORγt
Figure 2. SBI’s human and mouse RORγt lentivector and PiggyBac plasmids provide robust RORγt overexpression. The human and mouse RORγt lentivectors Cat.# TCL400A-1 (human) and Cat.# TCL300A-1 (mouse) and the PiggyBac transposon constructs Cat.# TCL400PB-1 (human) and Cat.# TCL300PB-1 (mouse) were transiently transfected into 293FT cells. After 24 hours, cellular lysates were prepared and Western blot analyses performed to detect the RORγt protein expression using an anti-RORγt antibody that detects both human and mouse proteins. Robust overexpression of both human (lanes 1,3) and mouse (lanes 2,4) RORγt proteins compared to the non-transfected cell lysate control (lane 5) can be clearly observed, with tubulin as a loading control.
Figure 3. SBI’s MSCV-Mouse RORγt-EF1α-GFP-T2A-Puro Jurkat Cell Line shows strong RORγt overexpression. Human Jurkat T cells were transduced with either Cat.# TCL400A-1 (human) or Cat.# TCL300A-1 (mouse) pre-packaged lentivirus and stable cell lines established using puromycin selection for seven days. Overexpression of RORγt was evaluated by measuring GFP levels using flow cytometry and Western blot analysis of cellular lysates and using anti-RORγt antibody. Both methods show that SBI’s RORγt-overexpressing Jurkat Cell Line possesses strong RORγt overexpression.