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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Mol Genet Metab. 2018 Apr 12;124(4):243–253. doi: 10.1016/j.ymgme.2018.04.001

Figure 1. Development of the Cps1cKO (tm1c) mouse.

Figure 1

The strategy for successful development of the mouse is diagrammed. 5′ LRPCR image and targeting confirmation of the F1 (tm1a) mice produced from the chimeric backcrossing: This incorporates a gene specific forward primer in the endogenous region upstream of the 5′ homology arm paired with a reverse primer in the lacZ cassette (in the en2/Frt junction). 3′ loxP image: This demonstrates the loxP confirmation of the targeted F1 mice produced from the chimeric backcrossing to ensure that the intact targeted allele has passed through the germline and proves it is the conditionally ready allele (tm1a). cKO (Tm1c) image: Screening of animals that are positive for the Flpo transgene demonstrates that successful recombination at the Frt sites has occurred, generating the conditional knockout (tm1c). This utilized a forward primer in the en2/Frt portion of the lacZ cassette and a reverse primer in the floxed region. With administration of AAV8-Cre recombinase, successful recombination at the loxP sites occurs, generating the knockout (tm1d). (LRPCR, long range polymerase chain reaction; DTA, Diphtheria Toxin A gene; Green ovals, FRT sites; red triangles, loxP sites; numbered black rectangles, exons)