Approximately 25% of the progeny from this mating will be homozygous for the loxP-flanked allele and hemizygous/heterozygous for the cre transgene. These will be your experimental mice.
Can Cre be homozygous?
Cre alone can produce a phenotype
Since hemizygous mice have a wildtype chromosome present, using hemizygous instead of homozygous Cre genotypes might minimize unintended consequences of random transgene insertion. … Cre can damage DNA in fibroblasts, gastric cells, and sperm in the absence of any floxed sequence.
How do you identify Cre expressions?
There are several ways to do this:
- Check for cre mRNA transcripts using quantitative RT-PCR or Northern analysis of mRNA harvested from the target tissue. …
- Evaluate Cre protein via immunohistochemistry (IHC) or by immunoblot.
Is Cre endogenous?
The Cre enzyme can be expressed in many different organisms such as plants, bacteria, mammals, yeast. … As mammalian genomes are on average in the region of 3×109 bp there is a very low chance of finding an endogenous loxP site. For Cre to be functional in a foreign host, exogenous loxP sites must be engineered.
Can you cross two Cre lines?
Given that Vav1-Cre is expressed in the ovaries and testes, there is no way to generate both wild-type (floxed) and mutant (cre-recombined) offspring with control littermates.
How was Cre lox discovered?
Sternberg discovered the recombinase, Cre, and its specific locus of crossover, lox, while studying the maintenance of bacteriophage P1 as a stable plasmid. Recombinations mediated by Cre assist in cyclization of the DNA of infecting phage and in resolution of prophage multimers created by generalized recombination.
Why is Cre recombinase so useful?
The Cre/loxP site-specific recombination system has emerged as an important tool for the generation of conditional somatic mouse mutants. … The spatial regulation of recombination can be achieved by using cell type-specific promoters that drive expression of Cre in the cell population or tissue of interest.
What is the meaning of Cre?
Carbapenem-resistant Enterobacterales (CRE) are multidrug-resistant organisms that that can cause serious infections and require interventions in healthcare settings to prevent spread.