PT  - JOURNAL ARTICLE
AU  - Tyler Joseph Dause
AU  - Elizabeth Diana Kirby
TI  - Poor concordance of floxed sequence recombination in single neural stem cells: Implications for cell autonomous studies
AID  - 10.1523/ENEURO.0470-19.2020
DP  - 2020 Feb 18
TA  - eneuro
PG  - ENEURO.0470-19.2020
4099  - http://www.eneuro.org/content/early/2020/02/18/ENEURO.0470-19.2020.short
4100  - http://www.eneuro.org/content/early/2020/02/18/ENEURO.0470-19.2020.full
AB  - To manipulate target gene function in specific adult cell populations, tamoxifen-dependent CreERT2 is widely used to drive inducible, site-specific recombination of loxP flanked sequences. In studies of cell autonomous target gene function, it is common practice to combine these CreERT2-lox systems with a ubiquitously expressed stop-floxed fluorescent reporter gene to identify single cells supposedly undergoing target gene recombination. Here, we studied the reliability of using Cre-induced recombination of one gene to predict recombination in another gene at the single cell level in adult hippocampal neural stem and progenitor cells (NSPCs). Using both probabilistic predictions in a generic experimental paradigm, as well as a mouse model with two separate stop-floxed reporters plus a Nestin promoter-driven CreERT2, we found that, in individual cells, recombination of one gene was a poor predictor of recombination in another. This poor concordance in floxed sequence recombination across genes suggests that use of stop-floxed reporters to investigate cell autonomous gene function may not be universally reliable and could lead to false conclusions.Significance Statement We investigate the reliability of a widely used transgenic mouse model in studies of adult NSPCs. Ligand-dependent Cre recombinases, such as the CreERT2 model, are a fundamental tool for inducible gene modification used to investigate gene function in many cell populations. It is common practice to combine NSPC-specific CreERT2-lox systems with a ubiquitously expressed stop-floxed fluorescent reporter gene to identify single cells undergoing target gene recombination in studies of cell autonomous gene function. Our probabilistic predictions and experimental data suggest that use of stop-floxed reporters to investigate cell autonomous gene function in NSPCs may lead to false conclusions because recombination in separate genes can show poor concordance in individual cells.