What is the Cre method?
The Cre-lox system is used as a genetic tool to control site specific recombination events in genomic DNA. The Cre protein is a site-specific DNA recombinase that can catalyse the recombination of DNA between specific sites in a DNA molecule.
What is the Cre lox system used for?
The Cre/lox system is one of the most powerful and versatile tools developed for mouse genetics. It gives mouse researchers sophisticated control over the location and timing of gene expression. Cre/lox is usually used to make knockout alleles, but it can also be used to activate gene expression.
Is Cre lox reversible?
Cre/loxP-mediated inversion of genomic DNA segments represents a useful tool to alter the mouse genome in a cell type- and/or tissue-specific manner. However, its applicability is hampered by the fact that this reaction is fully reversible.
What does CRE stand for?
CRE, which stands for carbapenem-resistant enterobacteriaceae, are strains of bacteria that are resistant to carbapenem, a class of antibiotics typically used as a last resort for treating severe infections when other antibiotics have failed.
How do you get CRE?
How are CRE germs spread? CRE are usually spread person to person through contact with infected or colonized people, particularly contact with wounds or stool (poop). This contact can occur via the hands of healthcare workers, or through medical equipment and devices that have not been correctly cleaned.
What is a Cre transgenic mouse?
A Cre mouse contains a Cre recombinase transgene under the direction of a tissue-specific promoter; a loxP mouse contains two loxP sites that flank a genomic segment of interest, the “floxed” locus. Typically, Cre and loxP mice are produced by using transgenic technology (Nagy 2000).
What is a Cre driver?
Cre-driver lines: A special type of transgenic mouse As shown in Figure 2 below, different types of neurons, whether inhibitory, excitatory or other have specific proteins and therefore genes that would allow for the specific expression of the exogenous Cre-recombinase.
What CRE precautions?
Wear a gown and gloves when caring for patients with CRE. Perform hand hygiene—use alcohol-based hand rub or wash hands with soap and water before and after contact with patient or their environment. Make sure labs immediately alert clinical and infection prevention staff when CRE are identified.
How is genetic code expansion used in Cre recombinase?
Genetic code expansion is used to replace a lysine residue critical to Cre recombinase function with a photocaged analogue of lysine, successfully blocking DNA recombination until irradiation with 405 nm light.
How is the expression of inducible Cre carried out?
Expression of inducible Cre uses the ER domain of the Cre-ER fusion protein to maintain the enzyme in the cell membrane until the receptor domain binds 4-hydroxytamoxifen, translocation to the cell nucleus occurs, and Cre recombinase acts on the site-specific recombinase sequences.
How is CRE used to generate conditional knockouts?
Both recombinases are highly efficient in excising DNA that lies in between the recognition sites and recombination requires only a minimal amount of recombinase expression. However, in order to generate conditional knockouts, temporal control of Cre is required.
How is double stranded DNA cut in Cre recombination?
Lox sites are directional and the two sites joined by the tetramer are parallel in orientation. The double stranded DNA is cut at both loxP sites by the Cre protein. The strands are then rejoined with DNA ligase in a quick and efficient process.
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