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In Situ Saturating Mutagenesis Screening Identifies a Functional Genomic Locus that Regulates Ucp1 Expression

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Abstract

A better understanding of the molecular mechanisms that control the UCP1 expression in brown and beige adipocytes is essential for us to modulate adipose cell fate and promote thermogenesis, which may provide a therapeutic view for the treatment of obesity and obesity-related diseases. To systematically identify cis-element(s) that transcriptionally regulates Ucp1, we here took advantage of the high-throughput CRIPSR-Cas9 screening system, and performed an in situ saturating mutagenesis screen, by using a customized sgRNA library targeting the ~ 20 kb genomic region near Ucp1. Through the screening, we have identified several genomic loci that may contain key regulatory element for Ucp1 expression in cultured brown and white adipocytes in vitro, and in inguinal white adipose tissue in vivo. Our study highlights a broadly useful approach for studying cis-regulatory elements in a high-throughput manner.

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All unique materials generated from this study are available from the corresponding author, Qiurong Ding (qrding@sibs.ac.cn).

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Funding

This work was supported by grants from the National Key R&D Program of China (2017YFA0102800, 2017YFA0103700), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16030402), and the National Natural Science Foundation of China (31670829, 31971063).

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Contributions

YQ and QD designed experiment. YQ performed all experiments. XL performed data analysis for CRIPSR screening. YS and SL assisted with the screening. YW and CT modified the pAAV-GFP plasmid. YQ and QD analyzed the data, prepared figures, and wrote the manuscript. QD supervised the project.

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Correspondence to Qiurong Ding.

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All authors declare no conflicts of interest.

Ethics Approval

All mice were housed under standard conditions at the Shanghai Institutes for Biological Sciences (SIBS), China. All animal procedures were performed according to guidelines, and were approved by the Institutional Animal Care and Use Committee of the Shanghai Institutes for Biological Sciences.

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Qiu, Y., Liu, X., Sun, Y. et al. In Situ Saturating Mutagenesis Screening Identifies a Functional Genomic Locus that Regulates Ucp1 Expression. Phenomics 1, 15–21 (2021). https://doi.org/10.1007/s43657-020-00006-7

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  • DOI: https://doi.org/10.1007/s43657-020-00006-7

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