5' Transgenes drive leaky expression of 3' transgenes in Cre-inducible bi-cistronic vectors.

ABSTRACT: Molecular cloning techniques enabling contemporaneous expression of two or more protein-coding sequences provide an invaluable tool for understanding the molecular regulation of cellular functions. The Cre-lox system is used for inducing the expression of recombinant proteins encoded within a bi-/poly-cistronic cassette. However, leak expression of transgenes is often observed in the absence of Cre recombinase activity, compromising the utility of this approach. To investigate the mechanism of leak expression, we generated Cre-inducible bi-cistronic vectors to monitor the expression of transgenes positioned either 5' or 3' of a 2A peptide or internal ribosomal entry site (IRES) sequence. Cells transfected with these bi-cistronic vectors exhibited Cre-independent leak expression specificall

SUBMITTER: Osanai Y 

PROVIDER: S-EPMC11298883 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature