2016 Fiscal Year Final Research Report
Search for new factors involved in glycolipid metabolism and transport by a genome-wide shRNA screen
| Project/Area Number |
26440069
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | National Institute of Infectious Diseases |
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Principal Investigator |
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Keywords | スフィンゴ糖脂質 / 志賀毒素 / ゲノムワイドスクリーニング / shRNA / ゲノム編集法 |
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| Outline of Final Research Achievements |
In this study, a genome-wide shRNA screen was performed using Shiga toxin (Stx)-induced cell death to search for new factors that are involved in glycolipid metabolism and transport. Several genes essential for the biosynthesis of Gb3, the Shiga-toxin receptor, were concentrated, which included glucosylceramide synthase. Furthermore, many shRNAs were concentrated in this screen. However, most of the Stx-resistant shRNAs showed off-target effects, which included an shRNA targeted for CERS2, one of the ceramide synthases. Then, CERS2 mutants were constructed by genome-editing, and non-resistance to Stx was confirmed in these mutants. On the other hand, sphingolipid biosynthesis was analyzed in these mutants, and the results suggested the existence of a yet-to-be-identified mechanism rendering very-long-chain-ceramides more accessible than C16-ceramide to glucosylceramide synthase, compared with sphingomyelin synthase.
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| Free Research Field |
糖鎖生物学 細胞生物学
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