Multiplex CRISPR/Cas9 editing of the long‐chain acyl‐CoA synthetase family in the diatom Phaeodactylum tricornutum reveals that mitochondrial ptACSL3 is involved in the synthesis of storage lipids

Autor: Xiahui Hao, Wenchao Chen, Alberto Amato, Juliette Jouhet, Eric Maréchal, Daniel Moog, Hanhua Hu, Hu Jin, Lingjie You, Fenghong Huang, Mark Moosburner, Andrew E. Allen, Yangmin Gong
Přispěvatelé: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences Ministry of Agriculture, Key Laboratory of Biology and Genetic Improvement of Oil Crops, LIPID, Physiologie cellulaire et végétale (LPCV), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Grenoble Alpes (UGA), Laboratory for Cell Biology, Philipps-University Marburg, Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Center for Microalgal Biotechnology and Biofuels, Institute of Hydrobiology, Chinese Academy of Sciences, Scripps Institution of Oceanography (SIO), University of California [San Diego] (UC San Diego), University of California-University of California, J. Craig Venter Institute [La Jolla, USA] (JCVI), National Natural Science Foundation of China (41776175 and 31961133008), Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences (CAAS-ASTIP-2013-OCRI), 2021 CEA-CAS bilateral program, Institut Carnot 3BCAR, Grants from the United States Department of Energy Genomics Science program, US National Science Foundation, NSF-MCB-1818390, Gordon and Betty Moore Foundation GBMF3828, ANR-10-LABX-0049,GRAL,Grenoble Alliance for Integrated Structural Cell Biology(2010), ANR-11-BTBR-0008,OCEANOMICS,Biotechnologies et bioressources pour la valorisation des écosystèmes marins planctoniques(2011), ANR-15-IDEX-0002,UGA,IDEX UGA(2015), Scripps Institution of Oceanography (SIO - UC San Diego), University of California (UC)-University of California (UC)
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: New Phytologist
New Phytologist, Wiley, 2021, ⟨10.1111/nph.17911⟩
New Phytologist, 2022, 233 (4), pp.1797-1812. ⟨10.1111/nph.17911⟩
ISSN: 0028-646X
1469-8137
Popis: International audience; Long-chain acyl-CoA synthetases (LACSs) play diverse and fundamentally important roles in lipid metabolism. While their functions have been well-established in bacteria, yeast and plants, the mechanisms by which LACS isozymes regulate lipid metabolism in unicellular oil-producing microalgae, including the diatom Phaeodactylum tricornutum, remain largely unknown. In P. tricornutum, a family of five genes (ptACSL1~5) encodes LACS activities. We generated single lacs knockout/knockdown mutants using multiplexed CRISPR/Cas9 method, and determined their substrate specificities towards different fatty acids (FAs) and subcellular localizations. ptACSL3 is localized in the mitochondria and its disruption led to compromised growth and reduced triacylglycerol (TAG) content when cells were bubbled with air. The ptACSL3 mutants showed altered FA profiles in two galactoglycerolipids and phosphatidylcholine (PC) with significantly reduced distribution of 16:0 and 16:1. ptACSL5 is localized in the peroxisome and its knockdown resulted in reduced growth rate and altered molecular species of PC and TAG, indicating a role in controlling the composition of acyl-CoAs for lipid synthesis. Our work demonstrates the potential of generating gene knockout mutants with the mutation of large fragment deletion using multiplexed CRISPR/Cas9 and provides insight into the functions of LACS isozymes in lipid metabolism in the oleaginous microalgae.
Databáze: OpenAIRE
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