title: pHUSH: a single vector system for conditional gene expression.
abstract: BACKGROUND
  Conditional expression vectors have become a valuable research tool to avoid artefacts that may result from traditional gene expression studies. However, most systems require multiple plasmids that must be independently engineered into the target system, resulting in experimental delay and an increased potential for selection of a cell subpopulation that differs significantly from the parental line. We have therefore developed pHUSH, an inducible expression system that allows regulated expression of shRNA, miRNA or cDNA cassettes on a single viral vector.
  
  
  RESULTS
  Both Pol II and Pol III promoters have been successfully combined with a second expression cassette containing a codon-optimized tetracycline repressor and selectable marker. We provide examples of how pHUSH has been successfully employed to study the function of target genes in a number of cell types within in vitro and in vivo assays, including conditional gene knockdown in a murine model of brain cancer.
  
  
  CONCLUSIONS
  We have successfully developed and employed a single vector system that enables Doxycycline regulated RNAi or transgene expression in a variety of in vitro and in vivo model systems. These studies demonstrate the broad application potential of pHUSH for conditional genetic engineering in mammalian cells.
authors: Department of Molecular Biology, Genentech Inc, South San Francisco, California, USA. Daniel.Gray@ucsf.edu
authors: Daniel C
authors: DC
authors: Gray
authors: Klaus P
authors: KP
authors: Hoeflich
authors: Li
authors: L
authors: Peng
authors: Zhenyu
authors: Z
authors: Gu
authors: Alvin
authors: A
authors: Gogineni
authors: Lesley J
authors: LJ
authors: Murray
authors: Mike
authors: M
authors: Eby
authors: Noelyn
authors: N
authors: Kljavin
authors: Somasekar
authors: S
authors: Seshagiri
authors: Mary J
authors: MJ
authors: Cole
authors: David P
authors: DP
authors: Davis
mesh_terms: D015870:Gene Expression / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics*; D014162:Transfection / Q000379:methods*
chemical_list: D011994:Recombinant Proteins