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Concept
Insertional mutagenesis
Résumé
In molecular biology, insertional mutagenesis is the creation of mutations in DNA by the addition of one or more base pairs. Such insertional mutations can occur naturally, mediated by viruses or transposons, or can be artificially created for research purposes in the lab.
Signature-tagged mutagenesis
This is a technique used to study the function of genes. A transposon such as the P element of Drosophila melanogaster is allowed to integrate at random locations in the genome of the organism being studied. Mutants generated by this method are then screened for any unusual phenotypes. If such a phenotype is found then it can be assumed that the insertion has caused the gene relating to the usual phenotype to be inactivated. Because the sequence of the transposon is known, the gene can be identified, either by sequencing the whole genome and searching for the sequence, or by using the polymerase chain reaction to amplify specifically that gene.
Because many viruses integrate their own genomes into the genomes of their host cells in order to replicate, mutagenesis caused by viral infections is a fairly common occurrence. Not all integrating viruses cause insertional mutagenesis, however.
Some DNA insertions will lead to no noticeable mutation. Historically, lentiviral vectors included strong viral promoters which had a side effect of insertional mutagenesis, nuclear DNA mutations that effect the function of a gene. These strong viral promotors were shown to be the main cause of cancer formation. As a result, viral promotors have been replaced by cellular promotors and regulatory sequences. For those viruses such as gammaretroviruses that tend to integrate their DNA in genetically unfavorable locations, the severity of any ensuing mutation depends entirely on the location within the host's genome wherein the viral DNA is inserted. If the DNA is inserted into the middle of an essential gene, the effects on the cell will be drastic. Additionally, insertion into a gene's promoter region can have equally drastic effects.
Source officielle
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