An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expression
Abstract Glucose, a primary fuel source under homeostatic conditions, is transported into cells by membrane transporters such as glucose transporter 1 (GLUT1). Due to its essential role in maintaining energy homeostasis, dysregulation of GLUT1 expression and function can adversely affect many physio...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-48361-5 |
| _version_ | 1827603918701461504 |
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| author | Yajuan Shi Ketaki A. Katdare Hyosung Kim Jonah C. Rosch Emma H. Neal Sidney Vafaie-Partin Joshua A. Bauer Ethan S. Lippmann |
| author_facet | Yajuan Shi Ketaki A. Katdare Hyosung Kim Jonah C. Rosch Emma H. Neal Sidney Vafaie-Partin Joshua A. Bauer Ethan S. Lippmann |
| author_sort | Yajuan Shi |
| collection | DOAJ |
| description | Abstract Glucose, a primary fuel source under homeostatic conditions, is transported into cells by membrane transporters such as glucose transporter 1 (GLUT1). Due to its essential role in maintaining energy homeostasis, dysregulation of GLUT1 expression and function can adversely affect many physiological processes in the body. This has implications in a wide range of disorders such as Alzheimer’s disease (AD) and several types of cancers. However, the regulatory pathways that govern GLUT1 expression, which may be altered in these diseases, are poorly characterized. To gain insight into GLUT1 regulation, we performed an arrayed CRISPR knockout screen using Caco-2 cells as a model cell line. Using an automated high content immunostaining approach to quantify GLUT1 expression, we identified more than 300 genes whose removal led to GLUT1 downregulation. Many of these genes were enriched along signaling pathways associated with G-protein coupled receptors, particularly the rhodopsin-like family. Secondary hit validation confirmed that removal of select genes, or modulation of the activity of a corresponding protein, yielded changes in GLUT1 expression. Overall, this work provides a resource and framework for understanding GLUT1 regulation in health and disease. |
| first_indexed | 2024-03-09T05:49:31Z |
| format | Article |
| id | doaj.art-ce1039e563874f36976d04d69d91cc82 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T05:49:31Z |
| publishDate | 2023-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-ce1039e563874f36976d04d69d91cc822023-12-03T12:19:00ZengNature PortfolioScientific Reports2045-23222023-11-0113111310.1038/s41598-023-48361-5An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expressionYajuan Shi0Ketaki A. Katdare1Hyosung Kim2Jonah C. Rosch3Emma H. Neal4Sidney Vafaie-Partin5Joshua A. Bauer6Ethan S. Lippmann7Department of Chemical and Biomolecular Engineering, Vanderbilt UniversityVanderbilt Brain Institute, Vanderbilt UniversityDepartment of Chemical and Biomolecular Engineering, Vanderbilt UniversityDepartment of Chemical and Biomolecular Engineering, Vanderbilt UniversityDepartment of Chemical and Biomolecular Engineering, Vanderbilt UniversityDepartment of Chemical and Biomolecular Engineering, Vanderbilt UniversityVanderbilt Institute of Chemical Biology, Vanderbilt UniversityDepartment of Chemical and Biomolecular Engineering, Vanderbilt UniversityAbstract Glucose, a primary fuel source under homeostatic conditions, is transported into cells by membrane transporters such as glucose transporter 1 (GLUT1). Due to its essential role in maintaining energy homeostasis, dysregulation of GLUT1 expression and function can adversely affect many physiological processes in the body. This has implications in a wide range of disorders such as Alzheimer’s disease (AD) and several types of cancers. However, the regulatory pathways that govern GLUT1 expression, which may be altered in these diseases, are poorly characterized. To gain insight into GLUT1 regulation, we performed an arrayed CRISPR knockout screen using Caco-2 cells as a model cell line. Using an automated high content immunostaining approach to quantify GLUT1 expression, we identified more than 300 genes whose removal led to GLUT1 downregulation. Many of these genes were enriched along signaling pathways associated with G-protein coupled receptors, particularly the rhodopsin-like family. Secondary hit validation confirmed that removal of select genes, or modulation of the activity of a corresponding protein, yielded changes in GLUT1 expression. Overall, this work provides a resource and framework for understanding GLUT1 regulation in health and disease.https://doi.org/10.1038/s41598-023-48361-5 |
| spellingShingle | Yajuan Shi Ketaki A. Katdare Hyosung Kim Jonah C. Rosch Emma H. Neal Sidney Vafaie-Partin Joshua A. Bauer Ethan S. Lippmann An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expression Scientific Reports |
| title | An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expression |
| title_full | An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expression |
| title_fullStr | An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expression |
| title_full_unstemmed | An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expression |
| title_short | An arrayed CRISPR knockout screen identifies genetic regulators of GLUT1 expression |
| title_sort | arrayed crispr knockout screen identifies genetic regulators of glut1 expression |
| url | https://doi.org/10.1038/s41598-023-48361-5 |
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