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A conditional gene knockout allows gene deletion in a tissue in a tissue specific manner. This is required in place of a gene knockout if the null mutation would lead to embryonic death, [13] or a specific tissue or cell type is of specific interest. This is done by introducing short sequences called loxP sites around the gene.
A knockout mouse (left) that is a model for obesity, compared with a normal mouse. There are several thousand different strains of knockout mice. [3] Many mouse models are named after the gene that has been inactivated.
A specific gene in mouse brain thought to be involved in the onset of Alzheimer's disease which codes for the enzyme cyclin-dependent kinase 5 (Cdk5) was knocked out. Such mice were found to be 'smarter' than normal mice and were able to handle complex tasks more intelligently compared to 'normal' mice bred in the laboratory.
Gene knock-in originated as a slight modification of the original knockout technique developed by Martin Evans, Oliver Smithies, and Mario Capecchi.Traditionally, knock-in techniques have relied on homologous recombination to drive targeted gene replacement, although other methods using a transposon-mediated system to insert the target gene have been developed. [3]
This figure depicts how Floxing is used in scientific research for spatial and temporal control of gene expression. In genetic engineering, floxing refers to the insertion of a DNA sequence (which is then said to be floxed) between two LoxP sequences, creating an artificial gene cassette which can then be conditionally deleted (knocked out), translocated, or inverted in a process called Cre ...
Vector containing DNA sequence similar to the gene to be modified is introduced to the cell, and by a process of recombination replaces the target gene in the chromosome. This method can be used to introduce a mutation or knock out a gene, for example as used in the production of knockout mice. [29]
Gene knockdown is an experimental technique by which the expression of one or more of an organism's genes is reduced. The reduction can occur either through genetic modification or by treatment with a reagent such as a short DNA or RNA oligonucleotide that has a sequence complementary to either gene or an mRNA transcript.
The gene can be later knocked out (inactivated) at a specific time-point or tissue-type in mutant mice derived from the mutant ES Cells, by appropriate breeding to other transgenic mice. This conditionality is a key property of the entire resource, and it allows a more nuanced study of the effects of the gene-knockout.
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