Cell lines for drug discovery: elevating target-protein levels using engineered transcription factors.
Drug discovery requires high-quality, high-throughput bioassays for lead identification and optimization. These assays are usually based on immortalized cell lines, which express the selected drug target either naturally or as a consequence of transfection with the cDNA encoding the target. Natural untransfected cell lines often fail to achieve the levels of expression required to provide assays of sufficient quality with a high enough signal-to-noise ratio. Unfortunately, the use of cDNA is increasingly restricted, as the sequences for more and more genes become subject to patent restrictions. To overcome these limitations, the authors demonstrate that engineered transcription factors with Cys2-His2 zinc finger DNA-binding domains can be used to effectively activate an endogenous gene of interest without the use of isolated cDNA of the target gene. Using this approach, the authors have generated a cell line that provides a high-quality and pharmacologically validated G-protein-coupled receptor bioassay. In principle, this technology is applicable to any gene of pharmaceutical importance in any cell type.
Sangamo BioSciences, Inc., Point Richmond Technology Center, Richmond, CA 94804, USA.
Pei-Qi
PQ
Liu
Magda F
MF
Morton
Andreas
A
Reik
Ragan
R
de la Rosa
Matthew C
MC
Mendel
Xiao-Yong
XY
Li
Casey C
CC
Case
Carl O
CO
Pabo
Veronica
V
Moreno
Ashley
A
Kempf
Jayashree
J
Pyati
Nigel P
NP
Shankley
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D015202:Protein Engineering; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D014157:Transcription Factors