Encyclopedia of Cancer

2011 Edition
| Editors: Manfred Schwab

Technical Knockout

  • Adi Kimchi
Reference work entry
DOI: https://doi.org/10.1007/978-3-642-16483-5_5711

Definition

Technical knockout (TKO) is a method for function-based gene isolation in mammalian cells.

Characteristics

Technical knockout application has resulted in the isolation of several novel genes involved in apoptosis and in the genetic dissection of certain processes within this important biological process.

In general, TKO is based on two concepts, both directed at overcoming impediments to the use of genetic screens in cultured cells.
  • One concept is the use of nucleic acids as “virtual mutagens” for trapping genes based on their function. This was achieved by transfections with antisense cDNA libraries to randomly inactivate gene expression, followed by a strong phenotypic selection.

  • The other concept was to develop efficient strategies for introduction and recovery of complex cDNA libraries, to allow multiple rounds of phenotypic selection and rapid screens for identification of functionally relevant genes. To this aim, an expression vector that provides an efficient gene...

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References

  1. 1.
    Kimchi A (1998) DAP genes: novel apoptotic genes isolated by a functional approach to gene cloning. Biochim Biophys Acta 1377:F13–F33PubMedGoogle Scholar
  2. 2.
    Deiss LP, Kimchi A (1991) A genetic tool used to identify thioredoxin as a mediator of a growth inhibitory signal. Science 252:117–120PubMedCrossRefGoogle Scholar
  3. 3.
    Deiss LP, Feinstein E, Berissi H et al (1995) Identification of a novel serine/threonine kinase and a novel 15-kD protein as potential mediators of the γ-interferon-induced cell death. Genes Dev 9:15–30PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2011

Authors and Affiliations

  1. 1.Department of Molecular GeneticsWeizmann Institute of ScienceRehovotIsrael