title: Knockin of Cre Gene at Ins2 Locus Reveals No Cre Activity in Mouse Hypothalamic Neurons.
abstract: The recombination efficiency and cell specificity of Cre driver lines are critical for exploring pancreatic β cell biology with the Cre/LoxP approach. Some commonly used Cre lines are based on the short Ins2 promoter fragment and show recombination activity in hypothalamic neurons; however, whether this stems from endogenous Ins2 promoter activity remains controversial. In this study, we generated Ins2-Cre knockin mice with a targeted insertion of IRES-Cre at the Ins2 locus and demonstrated with a cell lineage tracing study that the Ins2 gene is not transcriptionally active in the hypothalamus. The Ins2-Cre driver line displayed robust Cre expression and activity in pancreatic β cells without significant alterations in insulin expression. In the brain, Cre activity was mainly restricted to the choroid plexus, without significant recombination detected in the hippocampus or hypothalamus by the LacZ or fluorescent tdTomato reporters. Furthermore, Ins2-Cre mice exhibited normal glucose tolerance and insulin secretion upon glucose stimulation in vivo. In conclusion, this Ins2-Cre driver line allowed high-fidelity detection of endogenous Ins2 promoter activity in vivo, and the negative activity in the hypothalamus demonstrated that this system is a promising alternative tool for studying β cell biology.
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Kejia
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Xianhua
authors.initials: X
authors.lastname: Ma
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Xusheng
authors.initials: X
authors.lastname: Chang
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Shi
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.|Department of General Surgery, Changhai Hospital, Shanghai, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Yin
authors.affiliation: Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.|Department of Endocrinology, Changzheng Hospital, Shanghai, China.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Barshop Institute for Longevity and Aging Studies, University of Texas Health Science Center at San Antonio, San Antonio, TX 78245, USA.
authors.forename: Yuguang
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Zhifang
authors.initials: Z
authors.lastname: Xie
authors.affiliation: Department of Pathophysiology, Second Military Medical University, Shanghai 200433, China.|Center for Obesity &Diabetes Research and Innovation, Second Military Medical University, Shanghai 200433, China.
authors.forename: Weiping J
authors.initials: WJ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D004789:Enzyme Activation; D055879:Gene Knock-In Techniques; D023061:Gene Order; D018390:Gene Targeting; D056426:Genetic Loci; D005822:Genetic Vectors / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D059765:Homologous Recombination; D007031:Hypothalamus / Q000166:cytology* / Q000378:metabolism*; D007328:Insulin / Q000235:genetics* / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism; D019426:Integrases / Q000235:genetics* / Q000378:metabolism*; D007515:Islets of Langerhans / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000378:metabolism*; D010641:Phenotype
chemical_list: C561967:Ins2 protein, mouse; D007328:Insulin; C045073:Cre recombinase; D019426:Integrases; D005947:Glucose
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