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Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting mutant offspring (still retaining the frt-flanked neo) were bred to wildtype siblings, and maintained as such prior to sending heterozygous males to The Jackson Laboratory Repository in 2006.

Of note, the same Ch AT-IRES-Cre knock-in allele is also available as C57BL/6N-congenic (Stock No. An "IRES-Cre" sequence is inserted downstream of the stop codon such that cre expression is controlled by the endogenous Chat gene promoter. Cre recombinase activity is reported in all cholinergic neurons.

These mice may be useful for "Cre-lox" technology applications in neurobiology, including studies of motor function, learning and memory, Alzheimer's disease, and Down syndrome, and in obesity and diabetes research. A targeting vector was designed to insert an optimized internal ribosome entry site-linked Cre recombinase gene (followed by an frt-flanked neomycin resistance cassette) downstream of the stop codon of the endogenous gene.

Surprisingly, little is known of their breeding biology or reproductive success.

The North American breeding population has remained generally stable over the last 20 years, although some populations have declined in the eastern United States and along the Colorado River in the Southwest.

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