Gene knockdown

: "For knockdowns in combat sports, see Knockout." For|the song|Knockdown (song)

Gene knockdown refers to techniques by which the expression of one or more of an organism's genes is reduced, either through genetic modification (a change in the DNA of one of the organism's chromosomes) or by treatment with a reagent such as a short DNA or RNA oligonucleotide with a sequence complementary to either an mRNA transcript or a gene. If genetic modification of DNA is done, the result is a "knockdown organism". If the change in gene expression is caused by an oligonucleotide binding to an mRNA or temporarily binding to a gene, this results in a temporary change in gene expression without modification of the chromosomal DNA and is referred to as a "transient knockdown".

In a transient knockdown, the binding of this oligonucleotide to the active gene or its transcripts causes decreased expression through blocking of transcription (in the case of gene-binding), degradation of the mRNA transcript (e.g. by small interfering RNA (siRNA) or RNase-H dependent antisense) or blocking either mRNA translation, pre-mRNA splicing sites or nuclease cleavage sites used for maturation of other functional RNAs such as miRNAcite journal | last = Summerton | first = J | year = 2007 | title = Morpholino, siRNA, and S-DNA Compared: Impact of Structure and Mechanism of Action on Off-Target Effects and Sequence Specificity | journal = Med Chem. | volume = 7 |issue = 7 | pages = 651–660 | url = http://www.ncbi.nlm.nih.gov/pubmed/17430206 | format = Pubmed ] (e.g. by Morpholino oligos or other RNase-H independent antisensecite journal | last = Summerton | first = J | year = 1999 | title = Morpholino Antisense Oligomers: The Case for an RNase-H Independent Structural Type | journal = Biochimica et Biophysica Acta | volume = 1489 | pages = 141–58 | url = http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=10807004 | format = Pubmed ] ). The most direct use of transient knockdowns for learning about a gene that has been sequenced, but has an unknown or incompletely known function, an experimental approach known as reverse genetics. Researchers draw inferences from how the knockdown differs from individuals in which the gene of interest is operational. Transient knockdowns are often used in developmental biology because oligos can be injected into single-celled zygotes and will be present in the daughter cells of the injected cell through embryonic development [cite journal | last = Nasevicius | first = A | coauthors = Ekker SC | year = 2000 | title = Effective targeted gene 'knockdown' in zebrafish | journal = Nature Genetics | volume = 26 | issue = 2 | pages = 216–20 | url = http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11017081 | format = Pubmed | doi = 10.1038/79951 ] .

So far knockdown organisms with permanent alterations in their DNA have been engineered chiefly for research purposes. Also known simply as knockdowns, these organisms are most commonly used for reverse genetics, especially in species such as mice or rats for which transient knockdown technologies cannot easily be applied.

References

ee also

* Gene knockout