title: High-throughput siRNA-based functional target validation.
abstract: The drug discovery process pursued by major pharmaceutical companies for many years starts with target identification followed by high-throughput screening (HTS) with the goal of identifying lead compounds. To accomplish this goal, significant resources are invested into automation of the screening process or HTS. Robotic systems capable of handling thousands of data points per day are implemented across the pharmaceutical sector. Many of these systems are amenable to handling cell-based screening protocols as well. On the other hand, as companies strive to develop innovative products based on novel mechanisms of action(s), one of the current bottlenecks of the industry is the target validation process. Traditionally, bioinformatics and HTS groups operate separately at different stages of the drug discovery process. The authors describe the convergence and integration of HTS and bioinformatics to perform high-throughput target functional identification and validation. As an example of this approach, they initiated a project with a functional cell-based screen for a biological process of interest using libraries of small interfering RNA (siRNA) molecules. In this protocol, siRNAs function as potent gene-specific inhibitors. siRNA-mediated knockdown of the target genes is confirmed by TaqMan analysis, and genes with impacts on biological functions of interest are selected for further analysis. Once the genes are confirmed and further validated, they may be used for HTS to yield lead compounds.
authors.affiliation: Johnson & Johnson Pharmaceutical Research & Development LLC, Welsh and McKean Roads, Spring House, PA 19477-0776, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Xin
authors.forename: Alejandro
authors.initials: A
authors.lastname: Bernal
authors.forename: Frank A
authors.initials: FA
authors.lastname: Amato
authors.forename: Albert
authors.initials: A
authors.lastname: Pinhasov
authors.forename: Jack
authors.initials: J
authors.lastname: Kauffman
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Brenneman
authors.forename: Claudia K
authors.initials: CK
authors.lastname: Derian
authors.forename: Patricia
authors.initials: P
authors.lastname: Andrade-Gordon
authors.forename: Carlos R
authors.initials: CR
authors.lastname: Plata-Salamán
authors.forename: Sergey E
authors.initials: SE
authors.lastname: Ilyin
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D019295:Computational Biology; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D023281:Genomics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D034741:RNA, Small Interfering / Q000235:genetics*; D012371:Robotics; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering