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Thermo Scientific™ Jump In™ T-REx™ CHO-K1 Kit
Description
The Jump-In™ T-REx™ CHO-K1 Kit allows the targeted integration of genetic material into a specific pre-engineered R4 site in the Jump-In™ CHO-K1 cell line, to create an inducible isogenic stable cell line with less effort and in less time than traditional cell engineering methods. These cells stably express the tetracycline repressor protein, allowing for inducible expression of your retargeted gene of interest upon the addition of doxycycline.
The high retargeting efficiency, made possible by the R4 sites in the CHO-K1 cell line, allows the use of the isogenic pool for additional experiments without the need for clonal selection. Alternatively, the high retargeting efficiency allows for the easy selection of a positive stable clone expressing your gene of interest.
The Jump-In™ T-REx™ CHO-K1 Kit allows users to:
- Quickly and efficiently develop stably engineered isogenic cell pools in about half the time compared to traditional cell engineering methods
- Inducibly express your gene of interest using doxycycline
- Utilize isogenic expression from a defined genomic locus as the ideal solution for comparative analysis of gene families, isoforms, or orthologs
- Generate multiple cell lines in parallel using the simplified work flow
- Easily access the technology without complicated licenses or restrictions to interpret
Save Time with Rapid and Efficient Generation of Engineered Cell Lines
With the Jump-In™ T-REx™ CHO-K1 Kit you can generate functional cell pools in as little as 2 weeks without laborious clone isolation and analysis, and the streamlined workflow makes it easier to generate several cell lines at the same time. Even generation of clonal cell lines can be done with reduced time and effort due to the high percentage of positive clones. In addition, the Jump-In™ technology gives you the freedom to generate an unlimited number of cell lines without restrictive licensing requirements.
Expand Your Experimental Capabilities
The Jump-In™ T-REx™ CHO-K1 Kit allows for inducible expression of your toxic or unstable gene of interest and is the ideal solution for targets and assays where transient engineering technologies are problematic. The kit also provides a convenient way to create target panels of gene families, isoforms, or orthologs. Genes coding for large proteins or multi-unit proteins are not a problem since the Gateway™ destination vectors accept large inserts.
The kit includes:
- Jump-In™ T-REx™ CHO-K1 Cells, 2 x 1 mL vials
- pJTI™ R4 Dest CMV TO pA Vector, 100 μg—A Gateway™ Destination vector that drives inducible expression of your gene of interest under control of the strong CMV promoter by inclusion of the Tet-Operon.
- pJTI™ R4 Int Vector, 100 μg—Expresses the R4 integrase to facilitate the retargeting of your gene of interest into the R4 site.
Sold separately:
- pJTI™ R4 CMV-TO MCS pA Vector (Cat. No. A15004)—A multi-site cloning vector that drives inducible expression of your gene of interest under control of the strong CMV promoter by inclusion of the Tet-Operon.
- pJTI™ R4 EXP CMV-TO EmGFP pA Vector (Cat. No. A15005)—A control vector with inducible expression of GFP under control of the strong CMV promoter by inclusion of the Tet-Operon.
Specifications
Specifications
| Cell Line | CHO-K1 |
| Content And Storage | • Jump-In™ T-REx™ CHO-K1 Cells, 2 x 1 mL vials (store in liquid nitrogen) • pJTI™ R4 DEST CMV TO pA vector, 100 μg (store at -20°C) • pJTI™ R4 Int vector, 100 μg (store at -20°C) |
| Delivery Type | Transfection |
| Format | Kit |
| Promoter | CMV/TO |
| Vector | pJTI™ R4 DEST CMV TO pA , pJTI™ R4 Int |
| Expression System | Mammalian |
| For Use With (Application) | Assay development |
| Inducing Agent | Tetracycline |
| Key Functions | Stable Cell Line Development, Targeted Integration, Regulated Expression |
| Show More |
Frequently Asked Questions (FAQs)
When a co-transfection is performed, there is no way of testing the double stable cell line for functional TetR or GeneSwitch protein, respectively. On the other hand, when sequential transfection is performed, one can functionally test the generated T-REx or GeneSwitch cell line by transiently transfecting the lacZ expression control plasmid and then picking a clone that shows the lowest basal level of expression of lacZ in the absence of the inducer, and the highest level of lacZ in the presence of the inducer. This clone can then be expanded and used to transfect the T-REx or GeneSwitch expression construct, as the case may be.
With the GeneSwitch system, it is possible to have the absolute lowest basal levels of expression of the gene of interest, whereas the T-REx system may be a little leaky due to the inevitable presence of tetracycline in FBS. The induced level of expression in the GeneSwitch system can be even higher than that seen with the CMV promoter. The disadvantage of the GeneSwitch system is that the expression does not appear to switch off very easily in culture, although it has been demonstrated to function beautifully in transgenics. The T-REx system, on the other hand, can be switched on and off by the addition and removal of the inducer.
The Flp-In T-REx system combines the targeted integration offered by the Flp-In system with the powerful inducible expression offered by the T-REx system. It allows generation of isogenic, inducible, stable cell lines and permits polyclonal selection of these cell lines. Once the Flp-In T-REx host cell line containing an integrated FRT site has been created, subsequent generation of Flp-In T-REx cell lines expressing the gene(s) of interest is rapid and efficient.
Doxycycline may be used as an alternative inducing agent in the T-REx system. It is similar to tetracycline in its mechanism of action, and exhibits similar dose-response and induction characteristics as tetracycline in the T-REx system. Doxycycline has been shown to have a longer half-life than tetracycline (48 hours vs. 24 hours, respectively). We do not offer doxycycline, but it may be obtained from Sigma (Cat. No. D9891).
We offer three unique mammalian expression systems for inducible/regulated expression of the gene of interest:
- T-REx system
- Flp-In T-REx system
- GeneSwitch system
Please see below to see how they compare with one another:
System -- Basal Expression Level -- Induced Expression Level -- Response time to Maximal Expression -- Transgenic Appliation
T-Rex system -- Low -- Highest -- High -- Suitable
Flp-In T-REx system -- Lower -- High -- 24-48 hrs -- Suitable
GeneSwitch system -- Lowest -- High -- 24-48 hrs -- Suitable
For Research Use Only. Not for use in diagnostic procedures.
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