Gene knockout -Procedure, limitations and applications


  • Gene knockout is a one type of genetic technique that used in study structure and function of gene. Gene knock out also known as gene knockin. Gene knockout is a technique that also important in investigate the effect of gene.


  • Gene knockout technique are also known as gene replacement.
  • This technique was developed by capecchi in 1980.
  • This technique are important in the apply homologous recombination to changing in the genome of living organisms. this term also refers as a knocking out of gene.

    gene knockout
    Gene knockout

⇒ There are also other form of gene distribution.

  1. Gene knockdown that used in the reduce the statement of gene.
  2. Knock In that used in the replacement of one allele to another allele.
  • There are three types of gene knockout :-
  1. Double knockout

  2. Triple knockout

  3. Quadruple knockout

  • Knocking out of two genes its known as double knockout and in the same term knocking out of three gene its known as triple knockout.
  • Knocking out of four or five gene its known as quadruple knockout.


  • In the first step of this process we generate knocking out(CKO) targeting vector.
  • After the generate of vector we do electroporation of that vector into embrayonic stem(ES) cells.
  • We culture the embrayonic stem cells in fully laboratory condition at room temperature.
  • Then we pick a resistant colony from culture of embrayonic stem cells.
  • After we idenify embrayonic stem (ES) cells in southern blotting technique.
  • Sometimes low range of PCR are also used in the identification of embraonic stem cells.
  • Then we take targated embrayonic stem cell via through of microinjection technique.
  • Transfer of that stem cell into the uterus of pseudopregnent female mice.
  • After we identified chimeric cells in the uterus of mice.
  • Then we generate progeny that are carry the targeted allele.


  • Not each human disease can be identified by using of gene knockout technology.
  • No physical command on the this technology.
  • In gene knockout technology we do limited study on the embrayonic stem cells.
  • Not easy to find function of human genes in this technology.
  • Gene knockout technique also fail in make observable change in the mouse.
  • In gene knockout technology genetically ready embrayo can not grow into adult mice.
  • Reduction of adult mice is more difficult to find gene functions in humans health.


  • This technique are also important in the give important information about the functions of gene.
  • knockout technique information about the human genome as a genes are common between human and mice.
  • This technology is also help in study different model of cancer, obesity, heart disease, diabetes, arthritis, anxiety, and Parkinson disease.
  • They are also used in the study of gene products, biochemical pathways and developmental pathways.
  • Also important in study of different kinds of drug and therapies.
  • Also important in the modification of genome.
  • This technique also important information about mutations such as important in find deletion mutation and site specific mutations.
  • In this technique we also study about the recessive alleles.
  • Gene knockout technology also help in the study of functions of gene. Gene knockout also provides important information about mutations. Gene knockout technology are important in the made up of genome.
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