Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR /Cas9
Open accessAbstract
Summary: Microalgae are promising feedstock for biofuels yet mechanistic probing of their cellular network and industrial strain development have been hindered by lack of genome‐editing tools. Nannochloropsis spp. are emerging model microalgae for scalable oil production and carbon sequestration. Here we established a CRISPR /Cas9‐based precise genome‐editing approach for the industrial oleaginous microalga Nannochloropsis oceanica , using nitrate reductase ( NR ; g7988) as example. A new screening procedure that compares between restriction enzyme‐digested nested PCR ( nPCR ) products derived from enzyme‐digested and not‐digested genomic DNA of transformant pools was developed to quickly, yet reliably, detect genome‐engineered mutants. Deep sequencing of nPCR products directly amplified from pooled genomic DNA revealed over an 1% proportion of 5‐bp deletion mutants and a lower frequency of 12‐bp deletion mutants, with both types of editing precisely located at the targeted site. The isolated mutants, in which precise deletion of five bases caused a frameshift in NR translation, grow normally under NH 4 Cl but fail to grow under Na NO 3 , and thus represent a valuable chassis strain for transgenic‐strain development. This demonstration of CRISPR /Cas9‐based genome editing in industrial microalgae opens many doors for microalgae‐based biotechnological applications.
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Research and development efforts must focus on addressing gaps in current knowledge and innovation to overcome existing challenges.
“Research and development efforts must focus on addressing gaps in current knowledge and innovation to overcome existing challenges [31].”
From A Holistic Approach to Circular Bioeconomy Through the Sustainable Utilization of Microalgal Biomass for Biofuel and Other Value-Added Products (2024) · cites this paper as [31]
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- Authors
- Wang, Q., Lu, Y., Xin, Y., Wei, L., Huang, S., & Xu, J.
- Affiliations
- Chinese Academy of Sciences (CN)
- Single‐Cell Center CAS Key Laboratory of Biofuels and Shandong Key Laboratory of Energy Genetics Qingdao Institute of BioEnergy and Bioprocess Technology Chinese Academy of Sciences Qingdao Shandong 266101 China
- Corresponding author
- Xu, Jian
- Venue
- The Plant Journal
- Publisher
- Wiley
- Published
- 2016-10-18
- Volume
- 88
- Issue
- 6
- Pages
- 1071-1081
- ISSN
- 0960-7412 (ISSN-L)
- 1365-313X
- OpenAlex
- W2518075094
- Cited by
- 312
- References
- 63
- Citations by year
- 2026: 25 · 2025: 27 · 2024: 26 · 2023: 39 · 2022: 34 · 2021: 35 · 2020: 37 · 2019: 36 · 2018: 26 · 2017: 24 · 2016: 3
- FWCI
- 15.42
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The Plant Journal (publishedVersion)
- PubMed (publishedVersion)
- Institutional knowledge base (Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences) (submittedVersion)
- Funders
- National Natural Science Foundation of China (31425002, 91231205)
- Ministry of Science and Technology of the People's Republic of China (2012CB721101, 2012AA02A707)
- Chinese Academy of Sciences (KSZD‐EW‐Z‐017, ZDRW‐ZS‐2016‐3)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Keywords
- CRISPR/Cas9 genome editing, Nannochloropsis oceanica, oleaginous microalgae, nitrate reductase, deletion mutants, transgenic microalgae, chassis strain, microalgal biofuels, oil production, carbon sequestration, mutant screening, nested PCR
- MeSH terms
- Gene Editing, Polymerase Chain Reaction, Nitrate Reductase, Microalgae, CRISPR-Cas Systems
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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