title: The 14th Ile residue is essential for Leptin function in regulating energy homeostasis in rat.
abstract: LEPTIN (LEP) is a circulating hormone released primarily from white adipocytes and is crucial for regulating satiety and energy homeostasis in humans and animals. Using the CRISPR technology, we created a set of Lep mutant rats that carry either null mutations or a deletion of the 14(th) Ile (LEP(∆I14)) in the mature LEP protein. We examined the potential off-target sites (OTS) by whole-genome high-throughput sequencing and/or Sanger-sequencing analysis and found no OTS in mutant rats. Mature LEP(∆I14) is incessantly produced and released to blood at a much elevated level due to the feedback loop. Structure modeling of binding conformation between mutant LEP(∆I14) and LEPTIN receptor (LEPR) suggests that the conformation of LEP(∆I14) impairs its binding with LEPR, consistent with its inability to activate STAT3-binding element in the luciferase reporter assay. Phenotypic study demonstrated that Lep(∆I14) rats recapitulate phenotypes of Lep-null mutant rats including obesity, hyperinsulinemia, hepatic steatosis, nephropathy, and infertility. Compared to the existing ob/ob mouse models, this Lep(∆I14/∆I14) rat strain provides a robust tool for further dissecting the roles of LEP in the diabetes related kidney disease and reproduction problem, beyond its well established function in regulating energy homeostasis.
authors.affiliation: Tongji University, School of Life Sciences and Technology, 1239 Siping Road, Shanghai 200092, China.
authors.forename: Shuyang
authors.initials: S
authors.lastname: Xu
authors.affiliation: Tongji University, School of Life Sciences and Technology, 1239 Siping Road, Shanghai 200092, China.
authors.forename: Xianmin
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.affiliation: Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, 800 Dong Chuan Road, Shanghai 200240, China.
authors.forename: Youtian
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Department of Regenerative Medicine, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Jinping
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Tongji University, School of Life Sciences and Technology, 1239 Siping Road, Shanghai 200092, China.
authors.forename: Di
authors.initials: D
authors.lastname: He
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Department of Regenerative Medicine, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Department of Regenerative Medicine, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Lu
authors.affiliation: Department of Human Genetics, David Geffen School of Medicine, University of California Los Angeles, CA 90095 USA.
authors.forename: Guizhen
authors.initials: G
authors.lastname: Du
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Department of Regenerative Medicine, Tongji University School of Medicine, Shanghai 200065, China.|Department of Human Genetics, David Geffen School of Medicine, University of California Los Angeles, CA 90095 USA.
authors.forename: Youjin
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Tiancheng
authors.initials: T
authors.lastname: Liu
authors.affiliation: Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Guohui
authors.initials: G
authors.lastname: Ding
authors.affiliation: Tongji University, School of Life Sciences and Technology, 1239 Siping Road, Shanghai 200092, China.
authors.forename: Jiayu
authors.initials: J
authors.lastname: Chen
authors.affiliation: Tongji University, School of Life Sciences and Technology, 1239 Siping Road, Shanghai 200092, China.
authors.forename: Shaorong
authors.initials: S
authors.lastname: Gao
authors.affiliation: Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, 800 Dong Chuan Road, Shanghai 200240, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wu
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Department of Regenerative Medicine, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Xue
authors.affiliation: Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Yixue
authors.initials: Y
authors.lastname: Li
authors.affiliation: Tongji University, School of Life Sciences and Technology, 1239 Siping Road, Shanghai 200092, China.|Department of Human Genetics, David Geffen School of Medicine, University of California Los Angeles, CA 90095 USA.
authors.forename: Guoping
authors.initials: G
authors.lastname: Fan
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D064113:CRISPR-Cas Systems; D004195:Disease Models, Animal; D004734:Energy Metabolism; D005260:Female; D006706:Homeostasis; D007532:Isoleucine / Q000378:metabolism*; D020738:Leptin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008820:Mice, Obese; D008958:Models, Molecular; D009154:Mutation; D009765:Obesity / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D054411:Receptors, Leptin / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D000073336:Whole Genome Sequencing
chemical_list: D020738:Leptin; D054411:Receptors, Leptin; D007532:Isoleucine