Enhancing the accumulation of eicosapentaenoic acid and docosahexaenoic acid in transgenic Camelina through the <scp>CRISPR‐Cas9</scp> inactivation of the competing <scp><i>FAE1</i></scp> pathway

Han, Lihua; Haslam, Richard P.; Silvestre, Susana; Lu, Chaofu; Napier, Johnathan A.

doi:10.1111/pbi.13876

Cited by 14 publications

(7 citation statements)

References 10 publications

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“…Recently, a transgenic camelina strain was developed using multigene metabolic engineering to produce 12% DHA in seeds 49 . In addition, this line was crossed with CsFAE1 knockout camelina containing high ALA content in seeds, further increasing omega-3 FA content (ETA, EPA, DPA, and DHA) from 27 to 33% 51 . Therefore, the high-ALA camelina developed in this study can be used to produce omega-3 FAs such as EPA and DHA.…”

Section: Discussionmentioning

confidence: 99%

Physaria fendleri FAD3-1 overexpression increases ɑ-linolenic acid content in Camelina sativa seeds

Park

Choi

Kim

2023

Sci Rep

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Camelina (Camelina sativa) is an oil crop with a short growing period, resistance to drought and cold, low fertilizer requirements, and can be transformed using floral dipping. Seeds have a high content of polyunsaturated fatty acids, especially ɑ-linolenic acid (ALA), at 32–38%. ALA is an omega-3 fatty acid that is a substrate for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the human body. In this study, ALA content was further enhanced by the seed-specific expression of Physaria fendleri FAD3-1 (PfFAD3-1) in camelina. The ALA content increased up to 48% in T2 seeds and 50% in T3 seeds. Additionally, size of the seeds increased. The expression of fatty acid metabolism-related genes in PfFAD3-1 OE transgenic lines was different from that in the wild type, where the expression of CsFAD2 decreased and CsFAD3 increased. In summary, we developed a high omega-3 fatty acid-containing camelina with up to 50% ALA content by introducing PfFAD3-1. This line can be used for genetic engineering to obtain EPA and DHA from seeds.

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Section: Discussionmentioning

confidence: 99%

Physaria fendleri FAD3-1 overexpression increases ɑ-linolenic acid content in Camelina sativa seeds

Park

Choi

Kim

2023

Sci Rep

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“…Recently, a transgenic camelina strain that produces 12% DHA in seeds was developed through multigene metabolic engineering 38 . In addition, the total omega-3 FA content was increased to 33% by crossing the plants whose ALA was increased to 47% by knocking out csfae1 and the plants that had 27% omega-3 FA content including ETA, EPA, DPA, and DHA 40 . Camelina, with an ALA content of 50%, developed in this study can be used to produce omega-3 FAs.…”

Section: Discussionmentioning

confidence: 99%

Physaria fendleri FAD3-1 overexpression increases ɑ-linolenic acid content in camelina seeds

Park

Choi

Kim

2022

Preprint

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Camelina (Camelina sativa) is an oil crop with a short growing period, resistance to drought and cold, low fertilizer requirements, and can be transformed using floral dipping. Camelina seeds have a high content of polyunsaturated fatty acids, especially ɑ-linolenic acid (ALA) at 32–38%. ALA is an omega-3 fatty acid (FA) that is a substrate for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the human body. In this study, ALA content was further enhanced by seed-specific expression of Physaria fendleri (Pf) FAD3-1 in camelina. When PfFAD3-1 was introduced into camelina using the seed-specific glycinin promoter, ALA content increased by 48% in T2 seeds and 50% in T3 seeds. In addition, the weight and size of seeds increased. The expression of FA metabolism related genes in GlyP:PfFAD3-1 transgenic camelina was different than that in the wild type, in which the expression of Camelina sativa (Cs) FAD2 decreased and that of CsFAD3 increased. In summary, we developed a high omega-3 FA containing camelina with up to 50% ALA content by introducing PfFAD3-1. This line can be used for genetic engineering to obtain EPA and DHA from seeds.

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“…The inhibition intensity of CRISPRi system can reach 1000-fold mostly in prokaryotes [30]. CRISPR interference is also widely used in plants such as Nicotiana tabacum, Zea mays, and Arabidopsis thaliana (as shown in Table 1) [31][32][33][34][35][36]. CRISPR interference can be used as an alternative tool for RNAi.…”

Section: Cas9 and Dcas9mentioning

confidence: 99%

Recent Progress and Future Prospect of CRISPR/Cas-Derived Transcription Activation (CRISPRa) System in Plants

Ding

Luo

et al. 2022

Cells

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Genome editing technology has become one of the hottest research areas in recent years. Among diverse genome editing tools, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated proteins system (CRISPR/Cas system) has exhibited the obvious advantages of specificity, simplicity, and flexibility over any previous genome editing system. In addition, the emergence of Cas9 mutants, such as dCas9 (dead Cas9), which lost its endonuclease activity but maintains DNA recognition activity with the guide RNA, provides powerful genetic manipulation tools. In particular, combining the dCas9 protein and transcriptional activator to achieve specific regulation of gene expression has made important contributions to biotechnology in medical research as well as agriculture. CRISPR/dCas9 activation (CRISPRa) can increase the transcription of endogenous genes. Overexpression of foreign genes by traditional transgenic technology in plant cells is the routine method to verify gene function by elevating genes transcription. One of the main limitations of the overexpression is the vector capacity constraint that makes it difficult to express multiple genes using the typical Ti plasmid vectors from Agrobacterium. The CRISPRa system can overcome these limitations of the traditional gene overexpression method and achieve multiple gene activation by simply designating several guide RNAs in one vector. This review summarizes the latest progress based on the development of CRISPRa systems, including SunTag, dCas9-VPR, dCas9-TV, scRNA, SAM, and CRISPR-Act and their applications in plants. Furthermore, limitations, challenges of current CRISPRa systems and future prospective applications are also discussed.

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Enhancing the accumulation of eicosapentaenoic acid and docosahexaenoic acid in transgenic Camelina through the CRISPR‐Cas9 inactivation of the competing FAE1 pathway

Cited by 14 publications

References 10 publications

Physaria fendleri FAD3-1 overexpression increases ɑ-linolenic acid content in Camelina sativa seeds

Physaria fendleri FAD3-1 overexpression increases ɑ-linolenic acid content in Camelina sativa seeds

Physaria fendleri FAD3-1 overexpression increases ɑ-linolenic acid content in camelina seeds

Recent Progress and Future Prospect of CRISPR/Cas-Derived Transcription Activation (CRISPRa) System in Plants

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