Encyclopedia of Cancer

Living Edition
| Editors: Manfred Schwab

Technical Knockout

  • Adi Kimchi
Living reference work entry
DOI: https://doi.org/10.1007/978-3-642-27841-9_5711-2

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...

Keywords

Phenotypic Selection Lysosomal Protease Human Carcinoma Cell Line Episomal Vector Antisense cDNA 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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References

  1. Deiss LP, Kimchi A (1991) A genetic tool used to identify thioredoxin as a mediator of a growth inhibitory signal. Science 252:117–120CrossRefPubMedGoogle Scholar
  2. 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 9:15–30CrossRefPubMedGoogle Scholar
  3. Kimchi A (1998) DAP genes: novel apoptotic genes isolated by a functional approach to gene cloning. Biochim Biophys Acta 1377:F13–F33PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Department of Molecular GeneticsWeizmann Institute of ScienceRehovotIsrael