| Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR/Cas9 | |
Wang, Qintao1,2,3; Lu, Yandu1,2,4; Xin, Yi1,2; Wei, Li1,2; Huang, Shi1,2,3; Xu, Jian1,2,3
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| 2016-12-01 | |
| 发表期刊 | PLANT JOURNAL
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| 卷号 | 88期号:6页码:1071-1081 |
| 摘要 | 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. |
| 文章类型 | Article |
| 关键词 | Oleaginous Microalgae Nannochloropsis Spp. Clustered Regularly Interspaced Short Palindromic Repeat-cas System Genome Editing Nitrate Reductase Technical Advance |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| DOI | 10.1111/tpj.13307 |
| 关键词[WOS] | HOMOLOGOUS RECOMBINATION ; CHLAMYDOMONAS-REINHARDTII ; NITROGEN-DEPLETION ; BIOFUEL PRODUCTION ; NEUTRAL LIPIDS ; OCEANICA IMET1 ; GUIDE RNA ; EVOLUTION ; SEQUENCE ; REVEALS |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS研究方向 | Plant Sciences |
| 项目资助者 | National Natural Science Foundation of China(31425002 ; Synthetic Biology Program from Ministry of Science and Technology of China(2012CB721101 ; Biological Carbon Sequestration Program from Chinese Academy of Sciences(KSZD-EW-Z-017 ; 91231205) ; 2012AA02A707) ; ZDRW-ZS-2016-3) |
| WOS类目 | Plant Sciences |
| WOS记录号 | WOS:000393126200013 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qibebt.ac.cn/handle/337004/9166 |
| 专题 | 单细胞中心组群 |
| 作者单位 | 1.Chinese Acad Sci, Qingdao Inst BioEnergy & Bioproc Technol, CAS Key Lab Biofuels, Single Cell Ctr, Qingdao 266101, Shandong, Peoples R China 2.Chinese Acad Sci, Qingdao Inst BioEnergy & Bioproc Technol, Shandong Key Lab Energy Genet, Qingdao 266101, Shandong, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Hainan Univ, Coll Oceanol, State Key Lab Marine Resource Utilizat South Chin, Haikou 570228, Hainan, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Qintao,Lu, Yandu,Xin, Yi,et al. Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR/Cas9[J]. PLANT JOURNAL,2016,88(6):1071-1081. |
| APA | Wang, Qintao,Lu, Yandu,Xin, Yi,Wei, Li,Huang, Shi,&Xu, Jian.(2016).Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR/Cas9.PLANT JOURNAL,88(6),1071-1081. |
| MLA | Wang, Qintao,et al."Genome editing of model oleaginous microalgae Nannochloropsis spp. by CRISPR/Cas9".PLANT JOURNAL 88.6(2016):1071-1081. |
| 条目包含的文件 | 条目无相关文件。 | |||||
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