Regulatable adenovector harboring the GFP and Yamanaka genes for implementing regenerative medicine in the brain

LEHMANN, MARIANNE; CANATELLI-MALLAT, MARTINA; CHIAVELLINI, PRISCILA; MOREL, GUSTAVO R.; REGGIANI, PAULA C.; HEREÑÚ, CLAUDIA B.; GOYA, RODOLFO G.

GENE THERAPY

NATURE PUBLISHING GROUP

Lugar: Londres; Año: 2019

0969-7128

iological rejuvenation by partial cell reprogramming is an emerging avenue of research. In this context, regulatable pluripotency gene expression systems are the most widely used at present. We have constructed a regulatable bidirectional adenovector expressing the humanized green fluorescent protein (GFP) and oct4, sox2, klf4, and c-myc genes (known as the Yamanaka genes or OSKM). The OSKM genes are arranged as a bicistronic tandem (hSTEMCCA tandem), which is under the control of a Tet-Off bidirectional promoter that also controls the expression of the gFP gene. Separately, a constitutive cassette expresses the regulatory protein tTA. Vector DNA was transfected in HEK293 Cre cells, which were additionally infected with the helper adenovector H14, unable to package its DNA due to the Cre recombinase produced by the HEK293 Cre cells. The newly generated vector was expanded by six iterated coinfections of the above cells which were lysed at the end of the process and the adenovector pur