ArticlePlant biotechnology journal2021
Simultaneous knockout of multiple LHCF genes using single sgRNAs and engineering of a high-fidelity Cas9 for precise genome editing in marine algae.
Article in Plant biotechnology journal, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Roadmap for light interaction with biophotonic surfaces and their diverse applications.Journal of biomedical optics · 2026Article
- CRISPR-enhanced microalgae biosynthesis: a promising approach for future functional feed ingredients.Journal of animal science and biotechnology · 2026Review
- The marine diatom Phaeodactylum tricornutum as a versatile bioproduction chassis: Current progress, challenges, and perspectives.Plant communications · 2025Review
- Creating an Improved Diatoxanthin Production Line by Knocking OutMarine drugs · 2025Article
- A new tool for engineering Phaeodactylum tricornutum: the METE promoter drives both high expression and BThe Plant journal : for cell and molecular biology · 2025Article
- Influence of Light Regimes on Production of Beneficial Pigments and Nutrients by Microalgae for Functional Plant-Based Foods.Foods (Basel, Switzerland) · 2025Review
- Article
- Understanding the Impact of Nitrogen Availability: A Limiting Factor for Enhancing Fucoxanthin Productivity in Microalgae Cultivation.Marine drugs · 2024Article
- Clustered Regularly Interspaced Short Palindromic Repeat/CRISPR-Associated Protein and Its Utility All at Sea: Status, Challenges, and Prospects.Microorganisms · 2024Review
- An unexpected hydratase synthesizes the green light-absorbing pigment fucoxanthin.The Plant cell · 2023Article
- Marine phytoplankton downregulate core photosynthesis and carbon storage genes upon rapid mixed layer shallowing.The ISME journal · 2023Article
- Loss of CpFTSY Reduces Photosynthetic Performance and Affects Insertion of PsaC of PSI in Diatoms.Plant & cell physiology · 2023Article
- CRISPR-based bioengineering in microalgae for production of industrially important biomolecules.Frontiers in bioengineering and biotechnology · 2023Review
- Recent Advances in Improving Gene-Editing Specificity through CRISPR-Cas9 Nuclease Engineering.Cells · 2022Review
- Recent Progress on Systems and Synthetic Biology of Diatoms for Improving Algal Productivity.Frontiers in bioengineering and biotechnology · 2022Review
- Knockout ofJournal of genetics · 2022Article
- Simultaneous knockout of multiple LHCF genes using single sgRNAs and engineering of a high-fidelity Cas9 for precise genome editing in marine algae.Plant biotechnology journal · 2021Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The CRISPR/Cas9 system is an RNA-guided sequence-specific genome editing tool, which has been adopted for single or multiple gene editing in a wide range of organisms. When working with gene families with functional redundancy, knocking out multiple genes within the same family may be required to generate a phenotype. In this study, we tested the possibility of exploiting the known tolerance of Cas9 for mismatches between the single-guide RNA (sgRNA) and target site to simultaneously introduce indels in multiple homologous genes in the marine diatom Phaeodactylum tricornutum. As a proof of concept, we designed two sgRNAs that could potentially target the same six light-harvesting complex (LHC) genes belonging to the LHCF subgroup. Mutations in up to five genes were achieved simultaneously using a previously established CRISPR/Cas9 system for P. tricornutum. A visible colour change was observed in knockout mutants with multiple LHCF lesions. A combination of pigment, LHCF protein and growth analyses was used to further investigate the phenotypic differences between the multiple LHCF mutants and WT. Furthermore, we used the two same sgRNAs in combination with a variant of the existing Cas9 where four amino acids substitutions had been introduced that previously have been shown to increase Cas9 specificity. A significant reduction of off-target editing events was observed, indicating that the altered Cas9 functioned as a high-fidelity (HiFi) Cas9 nuclease.
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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.