ArticleThe Plant journal : for cell and molecular biology2015
A fluorescence-activated cell sorting-based strategy for rapid isolation of high-lipid Chlamydomonas mutants.
Article in The Plant journal : for cell and molecular biology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 96 citations in OpenAlex.
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- Label-free isolation of lipid-rich Saccharomyces cerevisiae mutant by high-throughput flow-mode Raman-activated cell sorting and multi-omics analysis for uncovering the mechanism of enhanced lipid accumulation.Biotechnology for biofuels and bioproducts · 2025Article
- Marine Phytoplankton Bioactive Lipids and Their Perspectives in Clinical Inflammation.Marine drugs · 2025Review
- Enhanced growth and metabolite production from a novel strain ofBioengineered · 2024Article
- The lifetime of the oxygen-evolving complex subunit PSBO depends on light intensity and carbon availability in Chlamydomonas.Plant, cell & environment · 2023Article
- Improved high-throughput screening technique to rapidly isolate Chlamydomonas transformants expressing recombinant proteins.Applied microbiology and biotechnology · 2022Article
- Enhancing carbohydrate repartitioning into lipid and carotenoid by disruption of microalgae starch debranching enzyme.Communications biology · 2021Article
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- Metabolic Engineering for Carotenoid Production Using Eukaryotic Microalgae and Prokaryotic Cyanobacteria.Advances in experimental medicine and biology · 2021Article
- Raman image-activated cell sorting.Nature communications · 2020Article
- A Versatile High Throughput Screening Platform for Plant Metabolic Engineering Highlights the Major Role ofFrontiers in plant science · 2020Article
- Bioengineering of Microalgae: Recent Advances, Perspectives, and Regulatory Challenges for Industrial Application.Frontiers in bioengineering and biotechnology · 2020Review
- Development and characterization of aBiotechnology for biofuels · 2020Article
- Rapid Enrichment and Isolation of Polyphosphate-Accumulating Organisms Through 4'6-Diamidino-2-Phenylindole (DAPI) Staining With Fluorescence-Activated Cell Sorting (FACS).Frontiers in microbiology · 2020Article
- Sheath-less high throughput inertial separation of small microparticles in spiral microchannels with trapezoidal cross-section.RSC advances · 2019Article
- Enhanced inertial focusing of microparticles and cells by integrating trapezoidal microchambers in spiral microfluidic channels.RSC advances · 2019Article
- Proteomic study uncovers molecular principles of single-cell-level phenotypic heterogeneity in lipid storage ofBiotechnology for biofuels · 2019Article
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- Glucose-Induced Trophic Shift in an Endosymbiont Dinoflagellate with Physiological and Molecular Consequences.Plant physiology · 2018Article
- Chlorophyll Fluorescence Video Imaging: A Versatile Tool for Identifying Factors Related to Photosynthesis.Frontiers in plant science · 2018Review
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Authors and funding
4 authors at 3 institutions in 1 country.
Funding
Abstract
There is significant interest in farming algae for the direct production of biofuels and valuable lipids. Chlamydomonas reinhardtii is the leading model system for studying lipid metabolism in green algae, but current methods for isolating mutants of this organism with a perturbed lipid content are slow and tedious. Here, we present the Chlamydomonas high-lipid sorting (CHiLiS) strategy, which enables enrichment of high-lipid mutants by fluorescence-activated cell sorting (FACS) of pooled mutants stained with the lipid-sensitive dye Nile Red. This method only takes 5 weeks from mutagenesis to mutant isolation. We developed a staining protocol that allows quantification of lipid content while preserving cell viability. We improved separation of high-lipid mutants from the wild type by using each cell's chlorophyll fluorescence as an internal control. We initially demonstrated 20-fold enrichment of the known high-lipid mutant sta1 from a mixture of sta1 and wild-type cells. We then applied CHiLiS to sort thousands of high-lipid cells from a pool of about 60,000 mutants. Flow cytometry analysis of 24 individual mutants isolated by this approach revealed that about 50% showed a reproducible high-lipid phenotype. We further characterized nine of the mutants with the highest lipid content by flame ionization detection and mass spectrometry lipidomics. All mutants analyzed had a higher triacylglycerol content and perturbed whole-cell fatty acid composition. One arbitrarily chosen mutant was evaluated by microscopy, revealing larger lipid droplets than the wild type. The unprecedented throughput of CHiLiS opens the door to a systems-level understanding of green algal lipid biology by enabling genome-saturating isolation of mutants in key genes.
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