Tuneable endogenous mammalian target complementation via multiplexed plasmidbased recombineering

Violeta Beltran-Sastre, Hannah Benistry, Julia Burnier, Imre Berger, Luis Serrano, Christina Kiel

Research output: Contribution to journalArticle (Academic Journal)

1 Citation (Scopus)
265 Downloads (Pure)

Abstract

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.
Original languageEnglish
Article number17432
Number of pages11
JournalScientific Reports
Volume5
DOIs
Publication statusPublished - 27 Nov 2015

Keywords

  • Diseases
  • Proteins

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