Gene Edited Crops in EU and EU policies
Fabien Nogué
Versailles Parma
Expert since 2012 in the GMO panel Molecular Characterization group Group leader at IJPB in Versailles
DNA Repair and Genome Engineering
Genetic variation: the basis of plant breeding
variety B modified
variety B
X
variety A
Ø The example of resistance to Phytophthora infestans that is the causal agent of blight in potato, the disease which led to the Great Irish Famine in 1845.
Ø The difference between the wild and cultivated potato is a genetic variation in the introgressed locus. One source of this genetic variation is mutagenesis.
Origin of genetic diversity
• Spontaneous mutations:
Around 10 double strand breaks occur per cell per day
Mutation rate per generation is 1 for 140 million base pairs in Arabidopsis.
wild cabbage
4 mutations between the mother plant and the seeds
DSBs
Genetic variation: the basis of plant breeding
- natural mutations are at the origin of variations -
Grapefruit red pulp, 1984 Cherry tree
self-fertility, 1985 Sunflower
high oleic acid, 1976
Ø Used in plant breeding since 70 years with more than 3200 varieties produced.
Origin of genetic diversity
• Induced mutations:
Mutation rate per generation is ≈ 1000 for 140 million base pairs in Arabidopsis with EMS.
Chemical or physical mutagens
Genetic variation: the basis of plant breeding
- induced mutations can be at the origin of variations -
X 1000
Ø Use of gene editing technology
ü modification of the target gene only
ü decrease the time for creation of new varieties
ü possibility to use genes of interest from other species
variety B modified
variety B variety or species A
Knowledge based identification of an allele of interest.
Ø mutant or QTL identification etc...
Ø Answers some limitations of classical breeding
ü risk of introgression of linked undesirable traits ü length of the vegetative cycle (trees ...)
ü difficulties of inter-species crosses and sexual compatibility barrier
Ø then break and repair the gene of interest
Origin of genetic diversity
• Targeted mutations:
Use of the CRISPR-Cas9 strategy.
Genetic variation: the basis of plant breeding
- targeted mutations can be at the origin of variations -
Ø Use of gene editing technology
ü modification of the target gene only
ü decrease the time for creation of new varieties
ü possibility to use genes of interest from other species
susceptible tomato wild potato
Knowledge based identification of an allele of interest.
Ø mutant or QTL identification etc...
Ø Answers some limitations of classical breeding
ü risk of introgression of linked undesirable traits ü length of the vegetative cycle (trees ...)
ü difficulties of inter-species crosses and sexual compatibility barrier
Ø then break and repair the gene of interest
Origin of genetic diversity
• Targeted mutations:
Use of the CRISPR-Cas9 strategy.
Genetic variation: the basis of plant breeding
- targeted mutations can be at the origin of variations -
resistant tomato
What is done in Europe on plant genome editing
Ø Examples of ini.a.ves on genome edi.ng and plant breeding innova.on in Europe
• France
: The GENIUS project “Genome engineering improvement for useful plants of a sustainable agriculture”. hAps://www6.inra.fr/genius-project_eng/Project/General-presentaHon• Netherland
: Call for projects 2017 by the Netherlands OrganisaHon for ScienHfic Research. Numerous projects on plant gene ediHng (University of Leiden ...).• Belgium
: Numerous projects at VIB Ghent with a first CRISPR/Cas9 Plant Field Trial in 2018.hAp://www.vib.be/en/training/research-training/courses/Pages/CRISPR-CAS-User-meeHng-2018.aspx
• England
: Opening of a CRISPR / Cas9 pla]orm by the JIC: wheat, barley, rapeseed, cabbage, potato, tomato ... hAps://www.jic.ac.uk/technologies/genomic-services/bract/• Italy
: Call for projects on plant genome ediHng planned in 2018 (more than 8M€)• Germany
: Call for projects 2017 (more than 6M€) NaHonal Strategy for Research in Bioeconomy 2030. Opening of a pla]orm for plant genome ediHng at the IPK of Gatersleben.• Acronyme
– Genome engineering improvement for useful plants of a sustainable agriculture
• Funding
– 7 years program (2012 to 2019) – Cost: 21,3 M€ (6 M€ from the ANR)
• Partnership
– 10 public and 4 private labs
– 11 life science and 3 social science labs
• Objec.ves
– Implement genome modificaKon techniques in crops
– Improve the efficiency and throughput of crop transformaKon – Provide proof of concept for improved traits useful for agriculture
What is done in France on plant genome editing
- The Genius project -
• 9 cul.vated plant species:
5 crops
2 vegetables 2 trees fruit and forestry
1 ornamental
Ø Genome ediHng is applicable to a
wide range of culHvated plant species
What is done in France on plant genome editing
Maize
SDN1
Potato
SDN1, BE
Tomato
SDN1, SDN2, BE
Wheat
SDN1
Rice
SDN1, SDN2
Rapeseed
SDN1
Apple tree
SDN1
Poplar
SDN1
Rose
SDN1
• 2 model species
Arabidopsis Physcomitrella
SDN1, SDN2, BE
Camelina
SDN1
Ø Genome edi.ng is a helpful tool for
• resistance to bioHc stress
• resistance to abioHc stress
• crop diversificaHon
• fast breeding
Resistance to the PVY virus in tomato
modification of the eIF4E gene
Salt stress No stress
Aeluropus liAoralis
Tolerance to the salt stress in rice
60% oleic acid
17% oleic acid
62%
Oil quality in the rustic crop camelina
modification of
the SAP9 gene Knock-out of the FAD2 gene
Knock-out of the TFL1 gene
Flowering time in apple tree
2 months plantlet 3-5 years for the
first flowers
What is done in France on plant genome editing
- A few examples from the Genius project -
General feeling about gene edited plants in EU
-before the ECJ ruling -
Ø Contrasted between countries and between stakeholders.
• Sweden (in 2015), Belgium and Netherland (in 2017), decided that the technical and legal arguments were sufficient to not regulate gene edited plants (field trials are in progress in VIB Belgium). France and in general Southern and Eastern European countries did not express any posiHons.
• EASAC (in 2017) advised EU regulators to regulate the specific agricultural trait/product rather than the technology by which it is produced.
• In France, in December 2015, 9 NGOs (including “ConfédéraHon
paysanne”) appealed to the French Conseil d’État to ask for a clarificaHon
on the status of gene edited plants. The Conseil d’État seized the Court of
JusHce of the European Union and the decision of the court arrived more
than 2 years later in July 2018.
Decision of the European Court of Justice
- the Ruling -
Ø The European Court of JusHce (ECJ) ruled on July 25, 2018, that:
“Organisms obtained by mutagenesis are GMOs and are, in principle, subject to the obligaKons laid down by the GMO DirecKve.
However, organisms obtained by mutagenesis techniques which have conven+onally been used in a number of applicaKons and have a long safety record are exempt from those obligaKons, on the understanding that the Member States are free to subject them, in compliance with EU law, to the obligaKons laid down by the direcKve or to other obligaKons.
”
hAps://curia.europa.eu/jcms/upload/docs/applicaHon/pdf/2018-07/cp180111en.pdf
GMO regulation in Europe
- definition -
Grapefruit red pulp, 1984 Cherry tree
self-fertility, 1985 Sunflower
high oleic acid, 1976
Ø Used in plant breeding since 70 years with more than 3200 varieties produced.
• Chemically or physically induced mutations, GMO but not regulated ...
“This DirecKve should not apply to organisms that have been obtained through
certain techniques of geneKc modificaKon which have conven+onally been
used in a number of applicaKons and have a long safety record.”
GMO regulation in Europe
- what about targeted mutagenesis -
Gene editing stable
transformation
Edited Plant
Back cross and selection Primary
transformant
Secondary transformant
Transient transformation
Edited Plant Gene
editing
CRISPR-Cas9
CRISPR-encoding transgene Targeted gene
Knock-out gene
Transient
Agrobacterium
Protoplasts Particule gun
via DNA
Stable
“This DirecKve should apply to organisms that have not been obtained through
certain techniques of geneKc modificaKon which have conven+onally been
used in a number of applicaKons and have a long safety record.”
GMO regulation in Europe
- back to the definition -
Protoplasts Particule gun
Edited Plant
via protein CRISPR-Cas9
CRISPR-encoding transgene Targeted gene
Knock-out gene Transient
transformation Gene editing
Ø “DNA free” edited plants are considered GMO and should be risk-assessed too.
“This DirecKve should apply to organisms that have not been obtained through
certain techniques of geneKc modificaKon which have conven+onally been
used in a number of applicaKons and have a long safety record.”
GMO regulation in Europe
- “long safety record” -
Ø Do transgenic plants have a long safety record now?
• First transgenic plant (soybean) in 1993 (25 years of safety record)
• First transgenic plant culHvated/imported (maize) to Europe in 1998 (20 years of safety record).
“This DirecKve should apply to organisms that have not been obtained through
certain techniques of geneKc modificaKon which have conven+onally been
used in a number of applicaKons and have a long safety record.”
GMO regulation in Europe
- “long safety record” -
Ø Why doing 1000 X more mutaHons would be more safe?
“This DirecKve should apply to organisms that have not been obtained through
certain techniques of geneKc modificaKon which have conven+onally been
used in a number of applicaKons and have a long safety record.”
Decision of the European Court of Justice
- Reactions to the Ruling -
Official Reac.ons
-• EU Commission: The Commission clarified recently that the Court decision is applicable immediately (edited plants are already
considered as GMOs in the EU).
• France: Government officially welcomed the clarificaHon of the status of gene edited plants. No further comments.
• Germany, more contrasted:
Ministry of Environment welcomed ruling,
Ministry of Research emphasized the risk for innovaHon.
• Other Member States – no official reacHons yet.
• USA: Agricultural secretary Perdue sent a clear message that he considers ECJ ruling not to be science-based.
• PoliHcal ParHes: Greens “ ECJ ruling is a victory for food
safety and the environment.”
Decision of the European Court of Justice
- Reactions to the Ruling - -
Stakeholders Reac.ons
• ConfédéraHon paysanne and other NGOs welcomed the ruling.
• Garlich von Essen, Secretary General of the European Seed AssociaHon said: “It is now likely that much of the potenHal of these innovaHve methods will be lost for Europe”.
• FNSEA, majority French agricultural union, said that ECJ ruling is a bad signal for agriculture in Europe.
• A number of scienHfic insHtuHons (Rothamsted, VIB, Wageningen, Max-Planck-InsHtute...) expressed their disappointment and
• more than 5000 scienHsts and ciHzens have signed a peHHon calling for “an immediate review of the ECJ ruling on mutagenesis”.
• posiHon paper from leading scienHsts represenHng more than 85 European plant and life sciences research centers.
• In UK, farming leaders and the UK agricultural industry joined scienHsts
to ask in an open leAer the UK government for clarity on gene-edited
crops.
Decision of the European Court of Justice
- Possible consequences -
Ø Defini.on of what is a GMO:
• It could blur the clear biological disHncHon between punctual mutaHons in an organism’s exisHng genes and the inserHon of very large secHons of recombinant DNA from a non-sexual compaHble species.
Ø Innova.on:
• Risk that these new rules could curb innovaHon in plant breeding in Europe.
• Costs associated with conducHng field trials under GMO regulaHons are extremely restricHve to research insHtutes and also to small biotech companies ($35.1 million for regulatory tesHng and registraHon process
EsHmate by Wendelyn Jones, Global Regulatory Affairs, DuPont Crop ProtecHon ).
Ø Trade:
• PosiHon of the EU could be uncomfortable towards the numerous countries that have already stated that they do consider crops made by simple gene ediHng in the same way as other forms of mutagenesis.
• Importers and port authoriHes will be in the complicated posiHon of policing food and feed grown as convenHonal crops in the countries of origin but that become illegal GMOs when they arrive in EU.
How to detect gene edited plants?
Ø Need for a method of detec.on:
• Standing CommiAee meeHng of the EU (September 2018) the JRC said:
- “How to detect something that can happen “naturally”?”
- ”the applicant has to develop the detecKon method”
- “when the modificaKon involves few nucleoKde changes, it would not be possible to iden+fy whether the mutaKon originated spontaneously or was induced by convenKonal or new (genome ediKng) mutagenesis techniques.”
- “it is unlikely that methods for the quanKficaKon of GMO products with small genome modificaKons in complex food or feed materials provide the level of selec+vity needed for the enforcement of legislaKon, such as the one on labelling.”
Joint Research Centre (JRC) European Commission's science and knowledge service provides independent scienHfic advice and support to EU policy.
Decision of the European Court of Justice
- Possible consequences -
Legal framework for GMO in Europe
- role of EFSA -
How to risk assess gene edited plants?
Ø In Europe risk assessment of GMO is done by EFSA:
• Agency of the European Union that carry out scienHfic risk assessment
on GMOs under two regulatory frameworks:
Decision of the European Court of Justice
- risk assessment of GMOs -
Decision of the European Court of Justice
- worst case scenario -
Ø Strict applicaHon of the actual guidelines and rules for GMO
Decision of the European Court of Justice
- Does every thing apply to gene editing -
Ø How was the CRISPR-Cas9 delivered? (transient versus stable, protein versus DNA). WGS analysis?
Ø Does the mutaHon already exist in this species?
• Should be true in most of the case .... History of safe use?
Decision of the European Court of Justice
- How to risk assess -
Ø How was the CRISPR-Cas9 delivered? (transient versus stable, protein versus DNA). NGS analysis?
Ø Does the mutaHon already exist in another species?
• Should be true in most of the case .... History of safe use?
Case of the eIF4E
gene found in pepper
and used in tomato.
Decision of the European Court of Justice
- An opportunity? -
PotenHally most of the quesHons will be on the funcHon rather than on the geneHc modificaHon and in many cases history of
safe use should apply.
variety B modified
variety B
Decision of the European Court of Justice
- Best case scenario -
Ø Product based rather than process based.
Ø Case by case and taking more into considera.on the history of safe use.
Opportunity for EFSA to set up an approach that would be more:
Ø More clarificaHons in few months ...
“The key is man's power of cumulative selection: nature gives successive variations; man adds them up in certain directions useful to him”.
(The Origin of Species, 1859)
“The power of Selection, whether exercised by man or brought into play under nature through the struggle for existence and the consequent survival of the fittest, absolutely depends on the variability of organic beings.
Without variability, nothing can be effected;”.
(Variation of Animals and Plants Under Domestication,1868)
Thank you for your attention
Charles Darwin
