New publication in Nucleic Acids Research

In the recently published NAR paper by Associate Eric Paul Bennett and colleagues, it is shown that the combined use of genome editing tools and last generation Tet-On systems can prevent leakiness and adverse effects associated with the lack of tight control of tetracycline-based inducible systems. 

The principle is based on precise integration of inducible transcriptional elements (coined PrIITE) targeted to: (i) exons of an endogenous gene of interest (GOI) and (ii) a safe harbour locus. Using PrIITE cells harboring a GFP reporter or CDX2 transcription factor, it was possible to demonstrate discrete inducibility of gene expression with complete abrogation of leakiness. CDX2 PrIITE cells generated by this approach uncovered novel CDX2 downstream effector genes. These results provide a strategy for characterization of dose-dependent effector functions of essential genes that require absence of endogenous gene expression.

The work was recently described in the UCPH Faculty of Health and Medical Sciences News. 

Pinto R, Hansen L, Hintze J, Almeida R, Larsen S, Coskun M, Davidsen J, Mitchelmore C, David L, Troelsen JT, Bennett EP (2017): Precise integration of inducible transcriptional elements (PrIITE) enables absolute control of gene expression. Nucleic Acids Res (Epub ahead of print)