Understanding the quantitative functional consequences of human disease mutations requires silencing of endogenous genes and expression of mutants at close to physiological levels. Changing protein levels above or below these levels is also important for system perturbation and modelling. Fast design optimization demands flexible interchangeable cassettes for endogenous gene silencing and tuneable expression. Here, we introduce ‘TEMTAC’, a multigene recombineering and delivery system for simultaneous siRNA-based knockdown and regulated mutant (or other variant) expression with different dynamic ranges. We show its applicability by confirming known phenotypic effects for selected mutations for BRAF, HRAS, and SHP2.
|Number of pages||11|
|Publication status||Published - 27 Nov 2015|
FingerprintDive into the research topics of 'Tuneable endogenous mammalian target complementation via multiplexed plasmidbased recombineering'. Together they form a unique fingerprint.
Professor Imre Berger
- School of Biochemistry - Professor of Biochemistry