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Conditional gene knockout is a technique used to eliminate a specific gene in a certain tissue, such as the liver. [ 1 ][ 2 ] This technique is useful to study the role of individual genes in living organisms. It differs from traditional gene knockout because it targets specific genes at specific times rather than being deleted from beginning ...
The Cre recombinase can recognize cryptic sites in the host genome and induce unauthorized recombination, damaging host DNA. This tool is suitable for deleting antibiotic resistance genes, but above all it allows conditional knockouts that can be induced at specific times in the cell type of choice.
The term "floxing" is a portmanteau constructed from the phrase "flanking/flanked by LoxP". The floxing method is essential in the development of scientific model systems as it allows researchers to have spatial and temporal alteration of gene expression. [ 2 ] The Cre-Lox system is widely used to manipulate gene expression in model organisms ...
Cre recombinase. Cre recombinase (Cre) is able to recombine specific sequences of DNA without the need for cofactors. The enzyme recognizes 34 base pair DNA sequences called loxP ("locus of crossover in phage P1"). Depending on the orientation of target sites with respect to one another, Cre will integrate/excise or invert DNA sequences.
Cre recombinase is a tyrosine recombinase enzyme derived from the P1 bacteriophage. The enzyme uses a topoisomerase I -like mechanism to carry out site specific recombination events. The enzyme (38 kDa) is a member of the integrase family of site specific recombinase and it is known to catalyse the site specific recombination event between two ...
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, [12] or a specific tissue or cell type is of specific interest. This is done by introducing short sequences called loxP sites around the gene.
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]
The Tet system has advantages over Cre, FRT, and ER (estrogen receptor) conditional gene expression systems. In the Cre and FRT systems, activation or knockout of the gene is irreversible once recombination is accomplished, whereas, in Tet and ER systems, it is reversible.