Characterization and Validation of Cre‐Driver Mouse Lines

Abstract

Conditional gene manipulations in mice are increasingly popular strategies in biomedical research. These approaches rely on the production of conditional genetically engineered mutant mouse (GEMM) lines with mutations in protein-encoding genes. These conditional GEMMs are then bred with one or several transgenic mouse lines expressing a site-specific recombinase, most often the Cre recombinase, in a tissue-specific manner. Conditional GEMMs can only be exploited if Cre transgenic mouse lines are available to generate somatic mutations, and thus the number of Cre transgenic lines has significantly increased over the last 15 years. Once produced, these transgenic lines must be validated for reliable, efficient, and specific Cre expression and Cre-mediated recombination. In this overview, the minimum level of information that is ideally required to validate a Cre-driver transgenic line is first discussed. The vagaries associated with validation procedures are considered next, and some solutions are proposed to assess the expression and activity of constitutive or inducible Cre recombinase before undertaking extensive breeding experiments and exhaustive phenotyping. Curr. Protoc. Mouse Biol. 1:1-15. © 2011 by John Wiley & Sons, Inc.

Identifying Essential Cell Types and Circuits in Autism Spectrum Disorders

“…In depth analyses of the sufficiency of these cell types to create ASD-like behavior disruptions are certainly needed. Determination of sufficiency in conditional deletion experiments must take into account that drivers of recombination have varying levels of specificity (Gofflot et al, 2011). Ultimately, converging lines of evidence, from multiple mouse models and human neuroanatomy will help to define the cell types and circuits which form the basis of ASD symptoms.…”

Atp6ap2 ablation in adult mice impairs viability through multiple organ deficiencies

“…To dissociate the general defects obtained with the ROSA-Cre deleter, we generated mice Atp6ap2 deletion specifically in the intestine using Villin-creERT2 mice (Atp6ap2 vil ). The murine Villin promoter provides stable and homogeneous expression of transgenes in small and large intestine along the crypt-villus axis, in differentiated enterocytes, as well as in the immature, undifferentiated cells of the crypt 34 . Four Atp6ap2 vil were injected with 1 mg of Tamoxifen for 5 days and were sacrificed 12 weeks after the last tamoxifen injection for the Atp6ap2 vilTAM mice.…”

“… 33 and the Tg( Villin-creERT2 ) allele, denoted here as Villin-CreERT2, was described in ref. 34 . After tamoxifen or control induction Atp6ap2 cKO/Y Tg( ROSA26-creERT2 ) animals are termed Atp6ap2 RosaTAM and Atp6ap2 RosaVEH , respectively.…”

Preserved dopaminergic homeostasis and dopamine‐related behaviour in hemizygous TH‐Cre mice

“…Importantly though, Cre‐driver lines might harbour aberrant phenotypes as a consequence of Cre‐induced abnormalities. High expression levels of Cre recombinase have been associated with cellular toxicity leading to compromised neuronal development (Forni et al ., ), significantly altered expression levels of endogenous proteins (Backman et al ., ) and integration‐mediated mutations or passenger‐genes (Lusis et al ., ), which all may cause aberrant phenotypes and confound the interpretation of observations (Gofflot et al ., ).…”