2022
DOI: 10.1186/s12915-022-01227-0
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A CRISPR toolbox for generating intersectional genetic mouse models for functional, molecular, and anatomical circuit mapping

Abstract: Background The functional understanding of genetic interaction networks and cellular mechanisms governing health and disease requires the dissection, and multifaceted study, of discrete cell subtypes in developing and adult animal models. Recombinase-driven expression of transgenic effector alleles represents a significant and powerful approach to delineate cell populations for functional, molecular, and anatomical studies. In addition to single recombinase systems, the expression of two recomb… Show more

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Cited by 14 publications
(10 citation statements)
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“…With type-specific marker genes identified with scRNA-seq, one can leverage an intersectional genetic strategy employing a dual recombinase system using both Cre recombinase as well as a second recombinase, flippase (Flp), to express the reporter genes in each specific LC neuronal type (or subtype). For this purpose, triple transgenic mice could be generated by crossing the Cre/Flp or Cre/Dre double-dependent reporters crossed with Dbh -cre (labeling all noradrenergic neurons) and an Flp driver line (expressing Flp in the marker gene locus) to label and manipulate each cell type (or subtype) while animals are trained to perform behavioral tasks (Lusk et al, 2022; Madisen et al, 2015).…”
Section: Discussionmentioning
confidence: 99%
“…With type-specific marker genes identified with scRNA-seq, one can leverage an intersectional genetic strategy employing a dual recombinase system using both Cre recombinase as well as a second recombinase, flippase (Flp), to express the reporter genes in each specific LC neuronal type (or subtype). For this purpose, triple transgenic mice could be generated by crossing the Cre/Flp or Cre/Dre double-dependent reporters crossed with Dbh -cre (labeling all noradrenergic neurons) and an Flp driver line (expressing Flp in the marker gene locus) to label and manipulate each cell type (or subtype) while animals are trained to perform behavioral tasks (Lusk et al, 2022; Madisen et al, 2015).…”
Section: Discussionmentioning
confidence: 99%
“…To validate the automated selection of breaths by BASSPRO, we compared the breaths selected by BASSPRO to those selected manually by experts in 16 representative traces from the 14 months timepoint in our Alzheimer’s dataset. Three experts (as defined by a record of peer-reviewed publication in respiratory waveform analysis 46 ) were engaged to select periods of calm, quiet breathing from regions of interest (i.e., stable gas concentrations for room air and challenge gas exposure) for analysis and comparison. The same files were then run through Breathe Easy using default parameters.…”
Section: Resultsmentioning
confidence: 99%
“…Essentially, this function takes as many points as are available for temperature and creates a linear interpolator that connects those measurements throughout the recording. This artificially provides body temperature measurements for the entire recording rather than two to three distinct temperatures copied throughout the recording as is currently applied 6, 34 . Currently, this interpolation cannot be turned off and is applied in all cases where temperature is not continuously recorded.…”
Section: Methodsmentioning
confidence: 99%
“…With type-specific marker genes identified with scRNA-seq, one can leverage an intersectional genetic strategy employing a dual recombinase system using both Cre recombinase as well as a second recombinase, flippase (Flp), to express the reporter genes in each specific LC neuronal type (or subtype). For this purpose, triple transgenic mice could be generated by crossing the Cre/Flp or Cre/Dre double-dependent reporters crossed with Dbh -cre (labeling all noradrenergic neurons) and an Flp driver line (expressing Flp in the marker gene locus) to label and manipulate each cell type (or subtype) while animals are trained to perform behavioral tasks ( Lusk et al, 2022 ; Madisen et al, 2015 ; Plummer et al, 2015 ).…”
Section: Discussionmentioning
confidence: 99%