Mup-knockout mice generated through CRISPR/Cas9-mediated deletion for use in urinary protein analysis
Haixia Yang,Wei Zhang,Shan Lu,Guangqing Lu,Hongjuan Zhang,Yinghua Zhuang,Wang Yue,Meng-Qiu Dong,Yu Zhang,Xingang Zhou,Peng Wang,Lei Yu,Fengchao Wang,Liang Chen +13 more
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TLDR
The successful knockout of the Mup gene cluster of 2.2 Mbp using the CRISPR/Cas9 system is reported, and a Mup-knockout mouse model is developed that will be useful for the research of urinary biomarker testing that may have relevance for humans.Abstract:
Major urinary proteins (MUPs) are the most abundant protein species in mouse urine, accounting for more than 90% of total protein content. Twenty-one Mup genes and 21 pseudogenes are clustered in a region of around 2 megabase pairs (Mbp) on chromosome 4. A Mup-knockout mouse model would greatly facilitate researches in the field of proteomic analysis of mouse urine. Here, we report the successful knockout of the Mup gene cluster of 2.2 Mbp using the CRISPR/Cas9 system. Homozygous Mup-knockout mice survived to adulthood and exhibited no obvious defects. The patterns of the proteomes of non-MUP urinary proteins in homozygous Mup-knockout mice were similar to those of wild-type mice judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The sensitivity of enzyme-linked immunosorbent assay to detect non-MUP urinary protein was significantly enhanced in Mup-knockout mice. In short, we have developed a Mup-knockout mouse model. This mouse model will be useful for the research of urinary biomarker testing that may have relevance for humans.read more
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Update on the human and mouse lipocalin ( LCN ) gene family, including evidence the mouse Mup cluster is result of an “evolutionary bloom”
Georgia Charkoftaki,Yewei Wang,Monica S. McAndrews,Elspeth A. Bruford,David C. Thompson,Vasilis Vasiliou,Daniel W. Nebert +6 more
TL;DR: Although much has been learned about LCNs and MUPs in recent years, more research is necessary to allow better understanding of their physiological functions, as well as their involvement in clinical disorders.
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Regulation of Sexually Dimorphic Expression of Major Urinary Proteins
TL;DR: A phylogenetic analysis on the origins of male-biased MUP gene expression in Mus musculus suggests that this sexual dimorphism evolved by increasing male MUP expression rather than reducing female expression.
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What if? Mouse proteomics after gene inactivation
Elisabetta Gianazza,Ingrid Miller,Uliano Guerrini,Luca Palazzolo,T. Laurenzi,Chiara Parravicini,Ivano Eberini +6 more
TL;DR: This review has gathered and organized all the available evidence and then compared the proteomic data in order to stress the context-specificity of the outcome every time two or more organs were investigated in the same KO mice.
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Urinary protein analysis in mice lacking major urinary proteins
TL;DR: In this paper, the MUP-knockout (Mup-KO) mice were produced by removing the Mup gene cluster using Cas9 proteins and two guide RNAs and characterized the urinary proteins in these mice.
Journal ArticleDOI
Comparative Proteomic Analysis of Liver Tissues and Serum in db/db Mice
TL;DR: The results show differentially expressed proteins (DEPs) in hepatic and serum proteomic analysis based on the leptin-receptor-deficient mouse model with overt obesity and NAFLD to offer sensitive non-invasive serum biomarkers for diabetes andNAFLD.
References
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The lipocalin protein family: structure and function
TL;DR: It is now clear that the lipocalins exhibit great functional diversity, with roles in retinol transport, invertebrate cryptic coloration, olfaction and pheromone transport, and prostaglandin synthesis.
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Identification of protein pheromones that promote aggressive behaviour
Pablo Chamero,Tobias F. Marton,Darren W. Logan,Kelly A. Flanagan,Jason R. Cruz,Alan Saghatelian,Benjamin F. Cravatt,Lisa Stowers +7 more
TL;DR: The results substantiate the idea of MUP proteins as pheromone ligands that mediate male–male aggression through the accessory olfactory neural pathway.
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sgRNAcas9: A Software Package for Designing CRISPR sgRNA and Evaluating Potential Off-Target Cleavage Sites
TL;DR: By identifying potential off-target sites in silico, the sgRNAcas9 allows the selection of more specific target sites and aids the identification of bona fide off- target sites, significantly facilitating the design of sg RNA for genome editing applications.
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Characterization of Genomic Deletion Efficiency Mediated by Clustered Regularly Interspaced Palindromic Repeats (CRISPR)/Cas9 Nuclease System in Mammalian Cells
Matthew C. Canver,Daniel E. Bauer,Abhishek Dass,Yvette Y. Yien,Jacky Chung,Jacky Chung,Takeshi Masuda,Takahiro Maeda,Takahiro Maeda,Barry H. Paw,Stuart H. Orkin +10 more
TL;DR: This work suggests that CRISPR/Cas9 is a robust system to produce a spectrum of genomic deletions to allow investigation of genes and genetic elements.
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Proteins in urine scent marks of male house mice extend the longevity of olfactory signals
TL;DR: The nature of their response suggests that, from a distance, mice may be unable to tell whether airborne signals emanate from scent marks or from the donor himself and it is suggested that this may provide territory owners with a major advantage in defending their territories.