By-Lovnish Thakur ASU2014010100099 Integrated Biotech 3rd Sem Course-Genetic Engineering Gmo's

KNOCKOUT MICE
By-Lovnish Thakur
ASU2014010100099
Integrated Biotech 3rd sem
Course-Genetic Engineering; Gmo’s
What are knockout mice?
A knockout mouse is a mouse in which a specific
gene has been inactivated or“knocked out” by
replacing it or disrupting it with an artificial
piece of DNA.
• The loss of gene activity often causes changes in

a mouse's phenotype and thus provides
valuable information on the function of the
gene.
Nobel Prize in Physiology in 2007
Mario R. Capecchi, Sir Martin J. Evans & Oliver

Smithies
"for their discoveries of principles for introducing

specific gene modifications in mice by the use of
embryonic stem cells".
Procedure
1. The gene to be knocked out is isolated from a
mouse gene library
2. Then a new DNA sequence is engineered which
is very similar to the original gene and its
immediate neighbor sequence, except that it is
changed sufficiently to make it inoperable.
3. From a mouse morula stem cells are isolated;
these can be grown in vitro. For this example, we
will take a stem cell from a white mouse.
4. The stem cells from step 3 are combined with the
new sequence from step 2.
• via electroporation
• selected using antibiotic
5. The stem cells from step 4 are inserted into mouse

blastocyst cells.
6. Blastocysts are then implanted into the uterus of
female mice, to complete the pregnancy.
7. The blastocysts contain two types of stem cells:
• The original ones
• The newly engineered ones
8. The newborn mice will therefore be chimeras:
parts of their bodies result from the original
stem cells, other parts result from the
engineered stem cells.
9. Newborn mice are only useful if the newly
engineered sequence was incorporated into the
germ cells (egg or sperm cells).
10. Cross these new mice with others and watch
for offspring that are all white. These are then
further inbred to produce mice that carry no
functional copy of the original gene.
Transgenic vs Knockout mice
A transgenic mouse typically
expresses one or more
copies of a gene (cDNA)that
is integrated into its genome
in a random fashion.
knockout mouse is a mouse

in which both alleles of a
gene are deleted in a
targeted fashion by
homologous recombination
Knockout vs Knockin
knockout in which a
gene or part of a gene is
deleted
Importance of knockout mice in
biology & medicine
1. provides valuable information about the function(s) of that gene.
2. genes are common between humans and mice, knockout mice gives
researchers information about the function of that gene in humans.
3. useful in studying and modeling different kinds of

• cancer,
• Obesity
• heart disease
• diabetes
• arthritis,
• anxiety,
• aging and
• Parkinson disease.
4. Knockout mice also offer a biological context in which drugs and other
therapies can be developed and tested
SOME EXAMPLES
A laboratory mouse in which a gene affecting hair growth has
been knocked out (left), is shown next to a normal lab mouse
A knockout mouse (left) that is a model of
obesity, compared with a normal mouse.
Limitations
• 15 percent of gene knockouts are developmentally lethal, which
means that the genetically altered embryos cannot grow into
adult mice.
• lack of adult mice- more difficult to determine a gene's function in

relation to human health.
• In some instances, the gene may serve a different function in

adults than in developing embryos.
• Knocking out a gene also may fail to produce an observable

change in a mouse
• There is variability in the whole procedure depending largely on

the strain from which the stem cells have been derived.
References
• http://www.cellmigration.org/resource/komo
use/komouse_approaches.shtml
• Cell Migration Consortium Transgenic &
Knockout Mouse Initiative
• http://learn.genetics.utah.edu/content/tech/t
ransgenic/
• http://www.eucomm.org/information/protoc
ols/
THANK YOU

By-Lovnish Thakur ASU2014010100099 Integrated Biotech 3rd Sem Course-Genetic Engineering Gmo's

Uploaded by

Copyright:

Available Formats

By-Lovnish Thakur ASU2014010100099 Integrated Biotech 3rd Sem Course-Genetic Engineering Gmo's

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

By-Lovnish Thakur ASU2014010100099 Integrated Biotech 3rd Sem Course-Genetic Engineering Gmo's

Uploaded by

Copyright:

Available Formats

KNOCKOUT MICE

• The loss of gene activity often causes changes in

Mario R. Capecchi, Sir Martin J. Evans & Oliver

"for their discoveries of principles for introducing

5. The stem cells from step 4 are inserted into mouse

knockout mouse is a mouse

3. useful in studying and modeling different kinds of

• lack of adult mice- more difficult to determine a gene's function in

• In some instances, the gene may serve a different function in

• Knocking out a gene also may fail to produce an observable

• There is variability in the whole procedure depending largely on

You might also like