Yes.

Oddly enough a genetic modification accomplished without human supervision for example an uncontrolled virus or fungus in the wild will not be classified as GMO.

In the human supervision there is an ethical standard not to take genetic source material from harmful organisms. Not so in the wild, there is no ethical standard among viruses and fungi.

Among friends there is an expectation that resistance to GMO will decrease after the patens expire.

Yes.

Oddly enough a genetic modification accomplished without human supervision for example an uncontrolled virus or fungus in the wild will not be classified as GMO.

In the human supervision there is an ethical standard not to take genetic source material from harmful organisms. Not so in the wild, there is no ethical standard among viruses and fungi.

Among friends there is an expectation that resistance to GMO will decrease after the patens expire.

In the US, products produced by the way described in your question, the USDA has no intention to regulate them. It was announced a few months back.

Given the technology associated in the process and the requirement for developing a product safe for consumption, one wish to have the CRISPR based products regulated.

CRISPR CAS-9 based products are obtained by editing and modifying genomes using the technology. In as much as the genome is modified with a desirable foreign DNA even though without bye product or after effect, the products are GMO.

@Godwin, I mean CRISPRs/Cas9 - caused mutation which changes an organism’s DNA without any foreign DNA introduced in the target plant genome - deletion made by guide RNA / Cas protein.

yes, I agree, the genome editing orgamisms are GMOs.

@Ruslan, Can you suggest any method to identify CRISPR/Cas9 edited plant with deletion of 3 nucleotides equal to a rare allele in the same species if such plant was obtained with guide RNA/Cas9 complex?

Alex, most people use sequencing technology to check the mutation, and see what is the size of the indel (deletion or insertion).

Yuan-Yeu Yau, I agree, I just wonder how we can differ natural allele from homologous deletion obtained via CRISPRs genome edition?

@Alex, you mean checking through 'phenotype', without using molecular analysis?

No, I mean accurate use of guide RNA /Cas 9 that delete a few nucleotides of the target gene. I know it sounds almost fantastic, but let's suggest that no other changes were found. How we can confirm GMO status of such plant if we know the same sequence with deletion in natural related plant? Besides the dossier of GM plant...

Genetically modified refers to plants and animals that have been altered in a way that wouldn’t have arisen naturally through evolution. A very obvious example of this involves transferring a gene from one species to another to endow the organism with a new trait. Meanwhile in CRISPR, genome editing technologies are used to introduce new plant traits by directly rewriting the plants’ genetic code.

Therefore, many countries like US wouldn’t regulate new plant varieties developed with new technologies like genome editing that would yield plants indistinguishable from those developed through traditional breeding methods. By contrast, a plant that includes a gene or genes from another organism, such as bacteria, is considered a GMO. This is another reason why many researchers and companies prefer using CRISPR in agriculture whenever it is possible.

Dear Mr. Ignatov,

this is a very interesting question.

Few days ago, the Court of Justice of the European Union (ECJ) has made a very important decision; it ruled that crops that are generated using CRISPR-CAS9 technology will be subjected to the European law that limited the planting and sale of genetically modified crops.

In my opinion, although CRISPR-CAS9 does not involve the introduction of foreign DNA and the sequential changes are not distinguishable from naturally occurring mutations, the "edited" organisms can be considered as genetically modified due to the fact that this is occurred upon human intervention.

However, basing on such statement, the use of other mutagenesis techniques involving chemicals and ionic irradiation that also induce genetic modifications without foreign DNA introduction should be evenly subjected to the same European restrictions.

Best regards

Mohammad

The term GMO is sometimes used by the public in a wider sense (like heterosis). What about to regenerate whole plants from cell cultures? Plants (I think, it is often done with potatoes) are more vigorous, but with high variation between single plants. I also heard, that this is due to loss of epigenetic information. GMO?

The Shouldn't be considered as GMOs

So long as they are free from DNA from foreign organisms I don't agree that they should be lablled as GMOs. Once the plants are edited with CRISPR/Cas9, we remove the T-DNA by crossing with WT plants and and screening for edited but non transgenic plants. Of course, the indels could range from a single nucleotide to several bps. However, in knock down there is no a single foreign DNA. In knock in cases there could be synthetic DNA. Now when we talk about GMOs it is general and doesn't discriminate how the organism has been developed. There needs to be a clear justification to accept or reject them. Since there are different ways of generating mutants with CRISPR/Cas technology the decision made by the European Union court, in my opinion is not justifiable. In plants I prefer to say Genetically Correceted Organisms (GCOs), since a single nucleotide can change the phenotype of the plant in question. If we delete few nucleotides from a specific target gene and develop a disease resistant plant, how many kgs/litres of chemicals could we avoid from polluting the environment, human beings and other animals? Environmentalists and policy/decision makers should on this issue for the benefit of mankind using this exception technology.

The problem is a different one. BECAUSE you are right, they will NOT. "How many kgs/litres of chemicals could we avoid from polluting the environment". This is "classical green". They KNOW those sentences. They are immune. Well, I am interested in plant resistance against the plantpathogen Xanthomonas. Virulence factors of this pathogen mimics eukaryotic transcription factors, but they dont bind exactly consensus sequences. As far as I know, very bad for citrus fruits and those factors are known, there are nearly allways central players ("pathogenicity factors") and it is known there they bind. Cut that genetic area out or modify it and the plant is resistant. I dont like that term "correction" (human genetics) that much. No one would diagree, if there wouldnt be a culture to do so.

Bananans without resistance against Xanthomonas, but a rice gene, which makes plants resistant. I am not against GMOs in general. Who would be, in that case? For many this crosses a line. And if they accept the argument against "correction", they hardly can argue in absolute fashion. That´s all. You are using the official strategy of argumentation. In that sense I agree with the critics: its not honest. - Can we really afford not to use GMOs?

I think it vary from situation to situation if the scientist just edit the plant genome it should not be categorized into GMO but if sequence of other organism is used to replace the original sequence it should be GMO

1. CRISPRs/Cas9 utilized vectors and constructs for delivery Cas9 and RNA to the cell. So, this is GMO.

2. Direct transfection CasX/RNA to the cell using Lipofectamine® 3000 - will produce multiple target DNA modification, because of complement to target 20 nt gRNA long is not unique enough to selectively editing. This is also GMO events.

CRISPR Gene Editing is a primitive tool and useless for application for eukaryotes.

My personal opinion, all enthusiastic fuss over this technology will soon come to an end:

CRISPR Gene Editing May Be Doing More Damage Than Scientists Thought

https://www.livescience.com/63075-crispr-damage-cell-mutations.html

A random cut

Ultimately, the precise consequences of these mutations are hard to estimate, as different types of cells use different operations to repair their DNA.

"Because you have a random repair process of rejoining DNA, I think anything is potentially possible when you're looking at billions of different events," Bradley said.

So, what does this mean for future CRISPR research? To Bradley, this study's findings should not discredit CRISPR-Cas9 as a promising genetic research tool but should make scientists cautious when thinking about using the gene-editing tool in a clinical setting.

Maria Jasin, a researcher from Memorial Sloan Kettering Cancer Center who was not involved in the study, agreed. "This study shows that further research and specific testing is needed before CRISPR-Cas9 is used clinically," Jasin said in a statement.

https://www.nature.com/articles/nbt.4192

Repair of double-strand breaks induced by CRISPR–Cas9 leads to large deletions and complex rearrangements

Michael Kosicki, Kärt Tomberg & Allan Bradley

Nature Biotechnology |

CRISPR–Cas9 is poised to become the gene editing tool of choice in clinical contexts. Thus far, exploration of Cas9-induced genetic alterations has been limited to the immediate vicinity of the target site and distal off-target sequences, leading to the conclusion that CRISPR–Cas9 was reasonably specific. Here we report significant on-target mutagenesis, such as large deletions and more complex genomic rearrangements at the targeted sites in mouse embryonic stem cells, mouse hematopoietic progenitors and a human differentiated cell line. Using long-read sequencing and long-range PCR genotyping, we show that DNA breaks introduced by single-guide RNA/Cas9 frequently resolved into deletions extending over many kilobases. Furthermore, lesions distal to the cut site and crossover events were identified. The observed genomic damage in mitotically active cells caused by CRISPR–Cas9 editing may have pathogenic consequences.

It must depend on its use for the purpose, and techniques involved to accomplish the job.

I don't totally agree with Ruslan Kalendar's idea

"CRISPR Gene Editing is a primitive tool and useless for application for eukaryotes."

I take it totally the opposite. For all of us in this forum, I think we need to differentiate between animals and plants. In animals, unless you use CRISPR/Cas complex to do its job in transient, the results might be as what has been said. However, in plants it is the state of the art technology. I agree that you can get different types of mutations in T0 or T1 generations. In some crops it seems that, once the CRISPR T-DNA is integrated in the genome there could be continuous activity and different types of mutations. In T0 plants to you can get plants with different mutations. However, it is so simple and easy to take out plants with T-DNA while you get edited plants. Through effective breeding you can finally get edited and non-transgenic plants, with only the specific area of our interest is only targeted. I suggest that, instead of generalizing better to separate between plants and animals.

I am very interested with this question. My idea is agree, but not agree with this technique. This technique is now very easy to use and could not detect whether or not it detect the gene from outside, if you do not see the procedure of gene construction and how to express in it. Important things is how this gene is expressed in the plant and whether or not it is safe for human, animals, and so on. This technique was publicated in 2012 by Dr. Doudna (Science). It is discussed all over the world. In Japan we are discussing about this question in the government and National Institutes. The Court of Justice of the European Union (ECJ) decided to prohibit this on January this year. I think this technique is not efficient for OK. One is to check on Off target. Last month I saw this paper in Nature Biotechnology. I think this technique is safety or not, it should be verified enough. I will wait for the direction of it. I will be searching for this article.

The discussion is amazing. I just wanted to ask two questions in connections to the topic:

Thanks

I appreciate the above valuable ideas from different aspects. But i don't think so we should mix the term of CRISPR/Cas9 with GMO. In GMO Plants we usually modify the genome with foreign material like Bt-cotton ( Strains of the bacterium Bacillus thuringiensis produce over 200 different Bt toxins, each harmful to different insects). What we do in CRISPR/Cas9?

I think very clear from all discussions.

Dear Tadesse!

As Malik said with GMOs we usually introduce a foreign gene to the organism and modify its genetic makeup, which might have several nucleotides from another source. In CRISPR/Cas 9 edited plants we expect at least the following phenomena:

1.....

i. Deletion of a single to several nucleotides

ii. Substitution of a single to a few nucleotides

iii. insertion of a few to several nucleotides to disrupt the function of an originally existing gene in that particular organism.

Therefore, CRISPR/Cas 9 edited plants could be considered as mutants with, mostly precise targets in the genome. Depending on the location of the specific gene you're targeting and the efficiency of you CRISPR/Cas9 machinery and other factors you can precisely target a specific gene and get results. I think these are similar to natural mutants, with precise editing.

2. In CRISPR/Cas 9 edited plants since we know our target gene. We can easily design our guide RNA and detect the situation with simple techniques, it is easily traceable and distinguishable from the GMOs with foreign genes (transgenic)? In flowering plants we can easily remove the T-DNA and create mutated but non-transgenic plants in about 2 years or less. I think there are may advantages of CRISPR for crop improvement. May be luck has also some contribution to be more successful......

Thank You Dr.Bekele Abebie for detail explanation of statement. I hope so such conversation will be very fruitful in understanding basic of science.

My additional Comments that may help to understand genome editing with CRISPR/Cas9.

In the following article published in Nature communications in 2017. The authors

described an efficient genome editing method for bread wheat using CRISPR/Cas9 ribonucleoproteins (RNPs).

https://www.nature.com/articles/ncomms14261

The authors claimed that they detected no off-target mutations in the mutant plants. "Because no foreign DNA is used in CRISPR/Cas9 RNP mediated genome editing, the mutants obtained are completely transgene free. This method may be widely applicable for producing genome edited crop plants and has a good prospect of being commercialized."

Such results are breakthrough technologies that can be used even in vegetatively propagated plants, like some banana varieties, where segregating out of the transgene is impossible.

Personally I totally disagree the ruling made by The EU Court of Justice that classed CRISPR edited plants to genetically modified plants. At least we need to discriminate the tow and need to follow different guidelines than the one used by EU. guidelines for GMOs.

What if we call CRISPR-MO (CMO), CMP (especially for plants) to indicate that they are developed by CRISPR/Cas9 technology and workout a new guideline for their approval.

May be the social scientists, who are the main decision makers, don't thoroughly understand (are biased by their advisors), to make such a ruling.

If you agree give your support.....

Thank you everybody, for useful discussion. I would assume, that:

1) Position of scientists on CRISPR-Cas genome edition is really affected by current situation with GMO in their own country.

2) There is a strong effect of intellectual rights on modern biotechnology methods owned by big biotech companies on public acceptance of GMO/Genome edited plants. Prof. Klaus Ammann (emeritus Director of Bern Botanic Garden) said it at 2003: "restriction for GMO is the most profitable for big biotech Co. - it keeps university researchers and small business out of this field".

3) in some cases (confirmed by genome sequencing) CRISPRs-edited plants could be similar to spontaneous mutants selected for useful trait (no off-target changes)

4) use of guide CRISPR RNA + Cas9 for several nucleotides deletion in the target genes does not introduce foreign DNA into plant genome and cannot be distinguished from natural mutation, but the method is not perfect so far.

Each and every technique has its own merits and demerits, safe or unsafe as far as its applications are concerned, but objective one has in his mind to achieve -using the technique -@drives to the category. CRISPR/cas9 is majorly used for genome editing so it has similar ststus as of spontaneous or induced mutation/s in the genomes. BUT if it is used to add and stack genes into a genome then definitely comes under GMO status thats what I think - you may disagree!

Many researchers and governments all over the world are now discussing on CRISPR-Cas9. I think depending on the circumstances one I might say 'Yes', the other might be 'No'. I think It is the same in the case of GMO. But I would say if a local citizens' group is opposed to this, you are not easy to get on 'Yes'. I think maybe governments also are same.

I agree with DR. Khan

As long as no foreign DNA of any kind has been inserted in the organism (except for mutations in the gene/genome, which are natural phenomena), I personally think that the organism should not be considered as GMO.

(The views expressed here are those of the individual only. It may not necessarily be the views of the institution/organization he is associated with).

agree with @Muhammad Sarwar Khan

Dear All

Thank you for your suggestions. I would agree to Alemayehu Teressa Negawo - it is not fair to treat differently new variety developed through conventional breeding, mutation and genetic engineering.

Happy New Year 2019!

Alex

In most cases, the aim of GMO is to introduce a new trait to the plant which does not occur naturally in the species ...... that is equal to genome editing (CRISPR).

Although CRISPR/CAS9 is just editing the same gene without insertion from different organism but in my personal opinion, CRISPR/CAS9 is the same in the case of GMO

Shatha

ı agree with Bekele Abebie opinions. Global world dealing with a lot of kinda stress for main nutritious plant so there is a tool for all of human being called CRISPR. I m not saying ragardless of consequences but under controlled enviroment desired trait could provied for us to nutritious food, oil, energy .... So using gene of interest for purpose changing exact locations could be CMO not GMO like sir bekele says.

Best regards.

UPDATE:

https://sciencebusiness.net/news/pressure-increases-ecj-rule-banning-use-genome-editing-agriculture-be-reversed