EU regulation of gene-edited plants—A reform proposal

Voigt, Brigitte

doi:10.3389/fgeed.2023.1119442

Cited by 9 publications

(5 citation statements)

References 9 publications

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“…However, under current legislation any deletion, even single base-pair deletions induced by CRISPR/Cas9 are still considered GMO in several areas of the world including the EU. However, deletions of genomic DNA induced by genome editing may be regulated as conventional plants in the future within EU countries [15,17]. This protocol may also be adapted for genome editing of other wild Solanum species.…”

Section: Discussionmentioning

confidence: 99%

A Walk on the Wild Side: Genome Editing of Tuber-Bearing Solanum bulbocastanum

Azariadis,

Andrzejczak,

Carlsen

et al. 2024

Plants

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Solanum bulbocastanum is a wild diploid tuber-bearing plant. We here demonstrate transgene-free genome editing of S. bulbocastanum protoplasts and regeneration of gene-edited plants. We use ribonucleoproteins, consisting of Cas9 and sgRNA, assembled in vitro, to target a gene belonging to the nitrate and peptide transporter family. Four different sgRNAs were designed and we observed efficiency in gene-editing in the protoplast pool between 8.5% and 12.4%. Twenty-one plants were re-generated from microcalli developed from individual protoplasts. In three of the plants we found that the target gene had been edited. Two of the edited plants had deletion mutations introduced into both alleles, whereas one only had a mutation in one of the alleles. Our work demonstrates that protocols for the transformation of Solanum tuberosum can be optimized to be applied to a wild Solanum species.

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Section: Discussionmentioning

confidence: 99%

A Walk on the Wild Side: Genome Editing of Tuber-Bearing Solanum bulbocastanum

Azariadis,

Andrzejczak,

Carlsen

et al. 2024

Plants

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“…These mutants are likely to have a higher level of customer acceptance and use of the biomass will be subject to fewer restrictions. Most countries outside the EU do not regulate transgene-free gene edited organism as GMOs [33], and the EU is currently establishing a new regulatory framework for this type of mutants [34]. Screening of transformants for CRISPR/Cas9induced mutations resulted in the identification of two zep2 and two zep3 knockout lines containing indels of different sizes causing frameshift mutations (Figure 2).…”

Section: Crispr/cas9-generated Zep2 and Zep3 Knockout Mutantsmentioning

confidence: 99%

<em>Zeaxanthin Epoxidase</em> 3 Knockout Mutants of the Model Diatom <em>Phaeodactylum tricornutum</em> Enable Commercial Production of the Bioactive Carotenoid Diatoxanthin

Græsholt,

Brembu,

Volpe

et al. 2024

Preprint

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Antioxidant, anti-inflammatory and chemo-preventive features have been reported for the carotenoid diatoxanthin. Diatoxanthin is only produced by a few groups of microalgae where it functions in photoprotection. Its large-scale production in microalgae is currently not feasible. In fact, rapid conversion into the inactive pigment diadinoxanthin is triggered when the cells are removed from the high-intensity light source, which will be the case during large-scale harvesting of microalgae biomass. Zeaxanthin epoxidase (ZEP) 2 and/or ZEP3 have been suggested to be responsible for the back-conversion of high-light accumulated diatoxanthin to diadinoxanthin in low light in diatoms. Using the CRISPR/Cas9 gene editing technology we knocked out the ZEP2 and ZEP3 genes in the already commercially used marine diatom Phaeodactylum tricornutum to investigate their role in the diadinoxanthin-diatoxanthin cycle, and to determine if one of the mutant strains could function as a diatoxanthin production line. Light shift experiments proved that ZEP3 encodes the enzyme converting diatoxanthin to diadinoxanthin in low light. Loss of ZEP3 caused the high-light accumulated diatoxanthin to be stable for several hours after the cultures had been returned to low light, suggesting that zep3 mutant strains could be suitable as commercial production lines of diatoxanthin.

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“…These mutants are likely to have a higher level of customer acceptance, and the use of the biomass will be subject to fewer restrictions. Most countries outside the EU do not regulate transgene-free gene-edited organisms as GMOs [33], and the EU is currently establishing a new regulatory framework for this type of mutant [34]. Screening of transformants for CRISPR/Cas9-induced mutations resulted in the identification of two zep2 and two zep3 knockout lines containing indels of different sizes causing frameshift mutations (Figure 3).…”

Section: Crispr/cas9-generated Zep2 and Zep3 Knockout Mutantsmentioning

confidence: 99%

Zeaxanthin epoxidase 3 Knockout Mutants of the Model Diatom Phaeodactylum tricornutum Enable Commercial Production of the Bioactive Carotenoid Diatoxanthin

Græsholt,

Brembu,

Volpe

et al. 2024

Marine Drugs

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Carotenoids are pigments that have a range of functions in human health. The carotenoid diatoxanthin is suggested to have antioxidant, anti-inflammatory and chemo-preventive properties. Diatoxanthin is only produced by a few groups of microalgae, where it functions in photoprotection. Its large-scale production in microalgae is currently not feasible. In fact, rapid conversion into the inactive pigment diadinoxanthin is triggered when cells are removed from a high-intensity light source, which is the case during large-scale harvesting of microalgae biomass. Zeaxanthin epoxidase (ZEP) 2 and/or ZEP3 have been suggested to be responsible for the back-conversion of high-light-accumulated diatoxanthin to diadinoxanthin in low-light diatoms. Using CRISPR/Cas9 gene editing technology, we knocked out the ZEP2 and ZEP3 genes in the marine diatom Phaeodactylum tricornutum to investigate their role in the diadinoxanthin–diatoxanthin cycle and determine if one of the mutant strains could function as a diatoxanthin production line. Light-shift experiments proved that ZEP3 encodes the enzyme converting diatoxanthin to diadinoxanthin in low light. Loss of ZEP3 caused the high-light-accumulated diatoxanthin to be stable for several hours after the cultures had been returned to low light, suggesting that zep3 mutant strains could be suitable as commercial production lines of diatoxanthin.

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EU regulation of gene-edited plants—A reform proposal

Cited by 9 publications

References 9 publications

A Walk on the Wild Side: Genome Editing of Tuber-Bearing Solanum bulbocastanum

A Walk on the Wild Side: Genome Editing of Tuber-Bearing Solanum bulbocastanum

<em>Zeaxanthin Epoxidase</em> 3 Knockout Mutants of the Model Diatom <em>Phaeodactylum tricornutum</em> Enable Commercial Production of the Bioactive Carotenoid Diatoxanthin

Zeaxanthin epoxidase 3 Knockout Mutants of the Model Diatom Phaeodactylum tricornutum Enable Commercial Production of the Bioactive Carotenoid Diatoxanthin

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