@article{c1e228ce5abe4e27bea8dbce5386c607,
title = "Knock-in mouse models for studying somatostatin and cholecystokinin expressing cells",
abstract = "Background: Somatostatin (SST) and cholecystokinin (CCK) are peptide hormones that regulate the endocrine system, cell proliferation and neurotransmission. New method: We utilized the novel Easi-CRISPR system to generate two knock-in mouse strains with Cre recombinase in SST- and CCK-expressing cells and validated their utility in the developing and adult brain tissues. Results: The full nomenclature for the newly generated strains are C57BL/6-Sstem1(P2A-iCre-T2A-mCherry)Mirn and C57BL/6-Cckem1(iCre-T2A-mCherry-P2A)Mirn. For the Sst locus, a P2A-iCre-T2A-mCherry cassette was inserted immediately upstream of the stop codon (C terminus fusion). For the Cck locus, iCre-P2A-mCherry-T2A cassette was inserted at the start codon (N terminus fusion). Knock-in mice were generated using the Easi-CRISPR method. Developmental and adult SST and CCK expressions were preserved and showed an appropriate expression pattern in both models, with an active fluorescent tag in both animal lines. Comparison with existing methods: Knock-in mouse models to study cell types that produce these critically important molecules are limited to date. The knock-in mice we generated can be used as reporters to study development, physiology, or pathophysiology of SST and CCK expressing cells – without interference with native expression of SST and CCK. In addition, they can be used as Cre driver models to conditionally delete floxed genes in SST and CCK expressing cells across various tissues. Conclusions: These two mouse models serve as valuable tools for in vitro and in vivo research studies related to SST and CCK biology across the lifespan and across different tissue types.",
keywords = "CRISPR/Cas9, Cholecystokinin, Cre, Easi-CRISPR, Somatostatin, ZsGreen1",
author = "Marta Balog and Allison Anderson and Gurumurthy, {Channabasavaiah B.} and Quadros, {Rolen M.} and Zeljka Korade and Karoly Mirnics",
note = "Funding Information: We are thankful to the UNMC DNA Sequencing Core Facility and the UNMC Comparative Medicine Department for their outstanding technical support of our experiments. This work was supported by National Institutes of Health (NIH), National Institues of Mental Health (NIMH), USA for R01 MH067234. CBG would like to acknowledge funding support from the National Institutes of Health, National Human Genome Research Institute, USA for R35HG010719; National Institute of General Medical Sciences for the R21 grant. The funding source had no influence on the study design, data collection or analyses, the decision to publish, or the preparation of this manuscript. The authors declare no conflict of interest. The datasets used and/or analyzed during the current study are available from the corresponding author within reasonable limits. Sst-Cre and Cck-Cre mice will be deposited into the Jackson Laboratories animal model repository immediately upon publication. Authors{\textquoteright} contributions: ZK and KM conceived and designed the study. CBG and RQ generated the mice. MB performed IHC and imaging. AA maintained the mouse colony, performed the genotyping, and assisted with tissue preparation for IHC. MB, AA, CG, RQ, ZK and KM interpreted the results and wrote the manuscript. All authors have read and given approval of the final version of the manuscript. Funding Information: We are thankful to the UNMC DNA Sequencing Core Facility and the UNMC Comparative Medicine Department for their outstanding technical support of our experiments. This work was supported by National Institutes of Health (NIH), National Institues of Mental Health (NIMH), USA for R01 MH067234 . CBG would like to acknowledge funding support from the National Institutes of Health, National Human Genome Research Institute , USA for R35HG010719 ; National Institute of General Medical Sciences for the R21 grant. The funding source had no influence on the study design, data collection or analyses, the decision to publish, or the preparation of this manuscript. The authors declare no conflict of interest. Publisher Copyright: {\textcopyright} 2022 Elsevier B.V.",
year = "2022",
month = nov,
day = "1",
doi = "10.1016/j.jneumeth.2022.109704",
language = "English (US)",
volume = "381",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
}