Rapid simultaneous cloning of drug targets from multiple mammalian species.
A method for the routine, rapid and simultaneous cloning of drug targets from multiple mammalian species is described. This expedites the generation of recombinant proteins and cell lines that can provide alternatives to animal experiments. This was achieved by the collection of RNA from a comprehensive range of tissues from a variety of species, and the optimisation of cDNA synthesis. This "zooplate" has been successfully used for the simultaneous amplification and cloning of drug targets from multiple species. These products have subsequently been used to develop in vitro assays that support efficacy and safety studies in new drug discovery programmes. Within the framework of the Three Rs, these reagents can reduce the number of animals required to provide material for ex vivo assays and can refine the in vivo studies that are still necessary.
Jupp
Rachael
R
Department of Molecular Biology, AstraZeneca R & D Charnwood, Loughborough, Leicestershire LE11 5RH, UK.
Dusanjh
Palminder K
PK
Walding
Andrew
A
McHale
Mark
M
Belfield
Graham P
GP
Delaney
Stephen J
SJ
D000826:Animal Testing Alternatives / Q000379:methods*; D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D012313:RNA / Q000737:chemistry / Q000235:genetics; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
C515187:CXCR3 protein, human; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine; D011994:Recombinant Proteins; D012313:RNA