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Related Experiment Videos

Virus-based expression systems facilitate rapid target in vivo functionality validation and high-throughput

Andrew L Darrow1, Kelly A Conway, Anil H Vaidya

  • 1Bioinformatics Group Leader, Johnson & Johnson Pharmaceutical Research & Development, Welsh and McKean Roads, Spring House, PA 19477, USA.

Journal of Biomolecular Screening
|July 12, 2003
PubMed
Summary

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This study presents a viral vector strategy for efficient drug discovery target validation and high-throughput screening (HTS). The method combines adenovirus gene transfer with baculovirus expression for rapid protein production and assay development.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Drug Discovery

Background:

  • Target validation is a critical bottleneck in modern drug discovery.
  • Efficient methods are needed to validate gene targets and develop high-throughput screening (HTS) assays.

Purpose of the Study:

  • To develop a generalized strategy combining viral gene transfer and expression systems for target validation and HTS assay development.
  • To demonstrate the utility of this approach for functional target validation and protein production.

Main Methods:

  • Adenovirus-mediated gene transfer for in vivo and in vitro target functionality validation.
  • Baculovirus expression system for producing sufficient protein quantities for HTS.
  • Incorporation of green fluorescent protein (GFP) for accelerated plaque purification.

Related Experiment Videos

  • Use of epitope and affinity tags for protein identification and purification.

    • Main Results:

    • Functional target validation was achieved in cell-based models and mouse cortex via adenovirus-mediated gene delivery.
    • Target overexpression led to the accumulation of a disease-related biomarker in vitro and in vivo.
    • A baculovirus system successfully produced adequate target protein for HTS.

    Conclusions:

    • The combined viral vector strategy offers a generalized method for rapid target validation and HTS assay development.
    • This approach is applicable to numerous gene targets identified in discovery programs.
    • The modular design facilitates seamless transition between target validation and HTS.