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- Publisher Website: 10.1111/tpj.13307
- Scopus: eid_2-s2.0-84991737543
- PMID: 27538728
- WOS: WOS:000393126200013
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Article: Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR/Cas9
Title | Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR/Cas9 |
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Authors | |
Keywords | clustered regularly interspaced short palindromic repeat-Cas system genome editing Nannochloropsis spp. nitrate reductase oleaginous microalgae technical advance |
Issue Date | 2016 |
Citation | Plant Journal, 2016, v. 88, n. 6, p. 1071-1081 How to Cite? |
Abstract | 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 NH4Cl but fail to grow under NaNO3, 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. |
Persistent Identifier | http://hdl.handle.net/10722/311537 |
ISSN | 2023 Impact Factor: 6.2 2023 SCImago Journal Rankings: 2.176 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Wang, Qintao | - |
dc.contributor.author | Lu, Yandu | - |
dc.contributor.author | Xin, Yi | - |
dc.contributor.author | Wei, Li | - |
dc.contributor.author | Huang, Shi | - |
dc.contributor.author | Xu, Jian | - |
dc.date.accessioned | 2022-03-22T11:54:10Z | - |
dc.date.available | 2022-03-22T11:54:10Z | - |
dc.date.issued | 2016 | - |
dc.identifier.citation | Plant Journal, 2016, v. 88, n. 6, p. 1071-1081 | - |
dc.identifier.issn | 0960-7412 | - |
dc.identifier.uri | http://hdl.handle.net/10722/311537 | - |
dc.description.abstract | 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 NH4Cl but fail to grow under NaNO3, 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. | - |
dc.language | eng | - |
dc.relation.ispartof | Plant Journal | - |
dc.subject | clustered regularly interspaced short palindromic repeat-Cas system | - |
dc.subject | genome editing | - |
dc.subject | Nannochloropsis spp. | - |
dc.subject | nitrate reductase | - |
dc.subject | oleaginous microalgae | - |
dc.subject | technical advance | - |
dc.title | Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR/Cas9 | - |
dc.type | Article | - |
dc.description.nature | link_to_OA_fulltext | - |
dc.identifier.doi | 10.1111/tpj.13307 | - |
dc.identifier.pmid | 27538728 | - |
dc.identifier.scopus | eid_2-s2.0-84991737543 | - |
dc.identifier.volume | 88 | - |
dc.identifier.issue | 6 | - |
dc.identifier.spage | 1071 | - |
dc.identifier.epage | 1081 | - |
dc.identifier.eissn | 1365-313X | - |
dc.identifier.isi | WOS:000393126200013 | - |