Polyswitch lentivectors: "all-in-one" lentiviral vectors for drug-inducible gene expression, live selection, and recombination cloning

Hum Gene Ther. 2011 Oct;22(10):1255-67. doi: 10.1089/hum.2010.179. Epub 2011 Sep 27.

Abstract

Lentiviral vectors are now widely considered one of the safest and most efficient tools for gene delivery and stable gene expression. Even though inducible gene expression cassettes are mandatory for many genetic engineering strategies, most current systems suffer from various issues, such as the requirement of two vectors, which decreases the overall efficiency of the transduction, leakiness and/or insufficient levels of activation of the inducible promoter, lack of selectable marker, low titers, or general issues associated with the cloning of large plasmids. In this article, we describe the design and functional characterization of a set of "all-in-one" multicistronic autoinducible lentivectors. They combine: (1) an optimized drug-inducible promoter; (2) a multicistronic strategy to express living color, selectable marker, and transactivator; and (3) acceptor sites for easy recombination cloning of genes of interest. These polyswitch lentivectors have good titers, very low basal activity, and reversible high induced activity, and can accept a growing number of genes already cloned in entry plasmids. These combined features make them a novel, powerful, and versatile tool for current and future genetic engineering approaches.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Toxins / genetics
  • Blotting, Western
  • DNA Primers / genetics
  • Gene Expression Regulation / genetics*
  • Gene Transfer Techniques*
  • Genetic Engineering / methods*
  • Genetic Vectors / genetics*
  • Lentivirus*
  • Promoter Regions, Genetic / drug effects
  • Promoter Regions, Genetic / genetics
  • RNA Splice Sites / genetics
  • Real-Time Polymerase Chain Reaction
  • Trans-Activators / genetics

Substances

  • Bacterial Proteins
  • Bacterial Toxins
  • CcdB protein, Plasmid F
  • DNA Primers
  • RNA Splice Sites
  • Trans-Activators