Precise integration of inducible transcriptional elements (PrIITE) enables absolute control of gene expression

• Tipo: Artigo em Revista Científica Internacional
• Data: 2017

Autores

Pinto, R (Autor) Outra; Hansen, L (Autor) Outra; Hintze, J (Autor) Outra; Raquel Almeida (Autor) FCUP; Larsen, S (Autor) Outra; Coskun, M (Autor) Outra; Davidsen, J (Autor) Outra; Mitchelmore, C (Autor) Outra; David L (Autor) FMUP; Troelsen, JT (Autor) Outra; Bennett, EP (Autor) Outra

Revista

Título: Nucleic Acids Research

Vol. 45

ISSN: 0305-1048

Editora: Oxford University Press

Indexação

ISI Web of Knowledge - 14 Citações

Scopus - 13 Citações

Outras Informações

ID Authenticus: P-00M-QZZ

DOI: 10.1093/nar/gkx371

Abstract (EN): Tetracycline-based inducible systems provide powerful methods for functional studies where gene expression can be controlled. However, the lack of tight control of the inducible system, leading to leakiness and adverse effects caused by undesirable tetracycline dosage requirements, has proven to be a limitation. Here, we report that the combined use of genome editing tools and last generation Tet-On systems can resolve these issues. Our 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 harbor locus. Using PrIITE cells harboring a GFP reporter or CDX2 transcription factor, we demonstrate discrete inducibility of gene expression with complete abrogation of leakiness. CDX2 PrIITE cells generated by this approach uncovered novel CDX2 downstream effector genes. Our results provide a strategy for characterization of dose-dependent effector functions of essential genes that require absence of endogenous gene expression.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 15