An easy method for preparation of Cre-loxP regulated fluorescent adenoviral expression vectors and its application for direct reprogramming into hepatocytes.

Date publication

décembre 2016

Journal

Biotechnology reports (Amsterdam, Netherlands)

Auteurs

Membres identifiés du Cancéropôle Est :
Dr WASYLYK Bohdan

Tous les auteurs :
Kurihara C, Nakade K, Pan J, Huang J, Wasylyk B, Obata Y

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/28352551

Résumé

The recombinant adenoviral gene expression system is a powerful tool for gene delivery. However, it is difficult to obtain high titers of infectious virus, principally due to the toxicity of the expressed gene which affects on virus replication in the host HEK293 cells. To avoid these problems, we generated a Cre-loxP-regulated fluorescent universal vector (termed pAxCALRL). This vector produces recombinant adenoviruses that express the red fluorescent protein (RFP) instead of the inserted gene during proliferation, which limits toxicity and can be used to monitor viral replication. Expression of the gene of interest is induced by co-infection with an adenovirus that expresses Cre-recombinase (AxCANCre). Recombinant adenovirus produced by this system that express Hnf4α and Foxa2 were used to reprogram mouse embryo fibroblast (MEF) into induced-hepatocyte-like cells (iHep) following several rounds of infection, demonstrating the efficacy of this new system.

Référence

Biotechnol Rep (Amst). 2016 Dec;12:26-32