Reference - Detail
| RRC ID | 78864 |
|---|---|
| Author | Tsuchiya M, Tachibana N, Nagao K, Tamura T, Hamachi I. |
| Title | Organelle-selective click labeling coupled with flow cytometry allows pooled CRISPR screening of genes involved in phosphatidylcholine metabolism. |
| Journal | Cell Metab |
| Abstract |
Cellular lipid synthesis and transport are governed by intricate protein networks. Although genetic screening should contribute to deciphering the regulatory networks of lipid metabolism, technical challenges remain-especially for high-throughput readouts of lipid phenotypes. Here, we coupled organelle-selective click labeling of phosphatidylcholine (PC) with flow cytometry-based CRISPR screening technologies to convert organellar PC phenotypes into a simple fluorescence readout for genome-wide screening. This technique, named O-ClickFC, was successfully applied in genome-scale CRISPR-knockout screens to identify previously reported genes associated with PC synthesis (PCYT1A, ACACA), vesicular membrane trafficking (SEC23B, RAB5C), and non-vesicular transport (PITPNB, STARD7). Moreover, we revealed previously uncharacterized roles of FLVCR1 as a choline uptake facilitator, CHEK1 as a post-translational regulator of the PC-synthetic pathway, and CDC50A as responsible for the translocation of PC to the outside of the plasma membrane bilayer. These findings demonstrate the versatility of O-ClickFC as an unprecedented platform for genetic dissection of cellular lipid metabolism. |
| Volume | 35(6) |
| Pages | 1072-1083.e9 |
| Published | 2023-6-6 |
| DOI | 10.1016/j.cmet.2023.02.014 |
| PII | S1550-4131(23)00050-5 |
| PMID | 36917984 |
| MeSH | CRISPR-Cas Systems / genetics Clustered Regularly Interspaced Short Palindromic Repeats* Flow Cytometry Lipid Metabolism* Organelles / metabolism Phosphatidylcholines / metabolism |
| IF | 21.567 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 121 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | K562(RCB0027) |