pLR: a lentiviral backbone series to stable transduction of bicistronic genes and exchange of promoters.
Gene transfer based on lentiviral vectors allow the integration of exogenous genes into the genome of a target cell, turning these vectors into one of the most used methods for stable transgene expression in mammalian cells, in vitro and in vivo. Currently, there are no lentivectors that allow the cloning of different genes to be regulated by different promoters. Also, there are none that permit the analysis of the expression through an IRES (internal ribosome entry site)-- reporter gene system. In this work, we have generated a series of lentivectors containing: (1) a malleable structure to allow the cloning of different target genes in a multicloning site (mcs); (2) unique site to exchange promoters, and (3) IRES followed by one of two reporter genes: eGFP or DsRed. The series of the produced vectors were named pLR (for lentivirus and RSV promoter) and were fairly efficient with a strong fluorescence of the reporter genes in direct transfection and viral transduction experiments. This being said, the pLR series have been found to be powerful biotechnological tools for stable gene transfer and expression.
Laboratório de Sinalização e Plasticidade Celular, Departamento de Biofísica, Universidade Federal do Rio Grande do Sul-UFRGS, 91501-970 Porto Alegre, RS, Brazil.
José Eduardo
JE
Vargas
Gabrielle
G
Salton
Andressa
A
Sodré de Castro Laino
Tiago Dalberto
TD
Pires
Martin
M
Bonamino
Guido
G
Lenz
Andrés
A
Delgado-Cañedo
D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014161:Transduction, Genetic; D019076:Transgenes
D049452:Green Fluorescent Proteins