Express challenging proteins in mammalian cells

Walkthrough: Comparison of three approaches to express challenging proteins in mammalian cells

In this video, we walk you through our poster, providing extra background and insights.

We are very excited to share this advance to our TARGATT™ large knock-in platform, as it enables applications in mammalian cells, such as expressing GPCRs and other membrane proteins in addition to toxic proteins.

If you’d like to download the poster, please complete the form on the left.

Frequently asked questions

Why isn’t there more induction when using pool-based enrichment for approach A?

Drug-based selection does not eliminate all parental cells, so some do not have the GOI donor. Also, a single non-toxic treatment with TAT-Cre does not reach all cells.

Why was there variation in the amount of induction when using clonal-isolation for approach A?

This is a consequence of the extent to which TAT-Cre treatments can reach cells. Also, these were all single tests (i.e. n=1 for each).

In what scenarios would the ~2% background expression and low induction efficiencies of approach B be acceptable?

Situations where large-scale inductions are needed, but TAT-Cre treatments are either not practical, or are not affordable.

Why are the induction efficiencies higher for approach C?

The GFP marker allows for tight exclusion of cells (via flow cytometry gating) that have not excised the stop element.