ArticleScientific reports2016
CRISPR/Cas9-induced knockout and knock-in mutations in Chlamydomonas reinhardtii.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 140 papers.
What it found
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Who cites it
140 citing papers in PubMed, 413 citations in OpenAlex.
- Review
- CRISPR-enhanced microalgae for carbon sequestration and carotenoid production: opportunities and fundamental constraints.Bioresources and bioprocessing · 2026Review
- CRISPR RNP-Mediated Transgene-Free Genome Editing in Plants: Advances, Challenges and Future Directions for Tree Species.Plant, cell & environment · 2026Review
- Light-dependent processes in algae across scales: from cellular physiology to ecological organization.Plant physiology · 2026Review
- Quantitative Evaluation of NR Locus-Targeted Nuclear Transformation inJournal of microbiology and biotechnology · 2026Article
- Enhancing CRISPR/Cas-Mediated Gene Knockout With Short Non-Homologous Oligonucleotides.Plant biotechnology journal · 2026Article
- Insights into transcriptomics and metabolic engineering of microalgal systems for enhancing industrial and environmental applications.World journal of microbiology & biotechnology · 2026Review
- Organelle-Targeted Multi-Enzyme Engineering Enables Medium-Chain Fatty Acid Production inJournal of microbiology and biotechnology · 2026Article
- Parametric Studies and Semi-Continuous Harvesting Strategies for Enhancing COMicroorganisms · 2026Article
- Metabolic Engineering of Microalgae for Biofuel Production.Methods in molecular biology (Clifton, N.J.) · 2026Review
- CRISPR-Cas Technology TurnsMarine drugs · 2025Review
- A quick-to-implement and optimized CRISPR-Cas9 protocol to obtain insertional and small indel mutants inMethodsX · 2025Article
- The marine diatom Phaeodactylum tricornutum as a versatile bioproduction chassis: Current progress, challenges, and perspectives.Plant communications · 2025Review
- Stable transgene expression and CRISPR-mediated knock-in system of a bacteria-derived antibiotic selection gene in the green alga Ulva prolifera.BMC plant biology · 2025Article
- Providing a toolbox for genomic engineering ofMicrobiology spectrum · 2025Article
- Transcriptional gene fusions via targeted integration at safe harbors for high transgene expression in Chlamydomonas reinhardtii.The New phytologist · 2025Article
- Beyond Cutting: CRISPR-Driven Synthetic Biology Toolkit for Next-Generation Microalgal Metabolic Engineering.International journal of molecular sciences · 2025Review
- Systematic approach for dissecting promoters and designing transform systems in microalgae.Microbial cell factories · 2025Article
- Genome editing in the green alga Chlamydomonas: past, present practice and future prospects.The Plant journal : for cell and molecular biology · 2025Review
- Cas9-mediated gene-editing frequency in microalgae is doubled by harnessing the interaction between importin α and phytopathogenic NLSs.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
80 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
20 authors at 5 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genome editing is crucial for genetic engineering of organisms for improved traits, particularly in microalgae due to the urgent necessity for the next generation biofuel production. The most advanced CRISPR/Cas9 system is simple, efficient and accurate in some organisms; however, it has proven extremely difficult in microalgae including the model alga Chlamydomonas. We solved this problem by delivering Cas9 ribonucleoproteins (RNPs) comprising the Cas9 protein and sgRNAs to avoid cytotoxicity and off-targeting associated with vector-driven expression of Cas9. We obtained CRISPR/Cas9-induced mutations at three loci including MAA7, CpSRP43 and ChlM, and targeted mutagenic efficiency was improved up to 100 fold compared to the first report of transgenic Cas9-induced mutagenesis. Interestingly, we found that unrelated vectors used for the selection purpose were predominantly integrated at the Cas9 cut site, indicative of NHEJ-mediated knock-in events. As expected with Cas9 RNPs, no off-targeting was found in one of the mutagenic screens. In conclusion, we improved the knockout efficiency by using Cas9 RNPs, which opens great opportunities not only for biological research but also industrial applications in Chlamydomonas and other microalgae. Findings of the NHEJ-mediated knock-in events will allow applications of the CRISPR/Cas9 system in microalgae, including "safe harboring" techniques shown in other organisms.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.