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Q:
Recent research shows that so-called "junk DNA" is in fact very significant in regulating genetic expression. Recent research also shows that specific conditions (e.g exercise vs non-exercise) affect DNA methylation and hence gene expression. 1) How do you know that the insertion point does not influence the function of the regulatory sections of the DNA? 2) How do you test the GMOs to make sure that the various environmental stressors (heat, drought, high humidity, fungi, bacteria, insects, pesticides, ....) do not inadvertently change genetic expression such that allergen or toxin production is increased? 3) Given the potential for multiple interactions among stressors, how do you control for that when you test for safety of the product?
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A:Expert Answer

Thanks for your question. Alan McHughen, CE Biotechnology Specialist and Geneticist recently wrote an article for GMO Answers that discusses inserted genes. The original article is posted at here, and is included below:

 

“Genes are like recipes, they tell the cell how to make a particular protein.  It is the presence (or absence) of the particular protein (often an enzyme) that gives the plant, animal or microbe a trait. Insulin, for example, is a protein that helps control blood sugar in mammals. The insulin gene recipe is carried in the genome of mammals, but is not present in the genomes of animals lacking blood, nor in plants or microbes, for that matter, as they have no blood to regulate.  Since the 1980s, insulin used by diabetics is made by Genetically Modified bacteria into which the human insulin gene recipe was inserted.  Although the bacteria have no insulin gene themselves, they were able to read and follow the human gene recipe to make insulin identical to the insulin made by human cells. This bacterial source GM insulin is extracted from the bacterial culture medium and provided to diabetics.

 

Genetic modification works only because all living things use the same genetic language, so a human gene—such as the insulin gene— transferred to bacteria will work the same way in the bacteria as it does in humans.

 

Genes are composed of long stretches of DNA, which is composed of the chemical building blocks we abbreviate as a,t,c and g (Adenine, Thymine, Cytosine and Guanine, respectively).  Just as in the human language English, in which out thousands of words are composed of specific sequences of 26 letters, in biology all genes in all species are made of specific sequences of the four DNA bases, a,t,c and g.  The human insulin gene consists of 4044 of these bases, beginning with …atggccctgtggatgcg… at the start of the insulin recipe.

 

When the plant, animal or microbe is ‘expressing’ a gene to make the requisite protein, it reads the DNA three letters at a time. Each three letter ‘word’ in DNA specifies a particular amino acid—of which there are twenty different kinds floating around in the cell—, and these amino acids serve as building block ingredients for proteins.  As the gene sequence is read by the cell’s kitchen machinery, amino acids are strung together like beads on a string, called a polypeptide chain. Thus, the first part of the DNA recipe, atg gcc ctg tgg atg cgc translates to the amino acids (in order) Methionine, Alanine, Leucine, Tryptophan, Methionine, Arginine, and so on until the entire sequence of 102 amino acids is completed. The string of amino acids is processed in the cell to release functional insulin.

 

Therefore, when engineers insert a known DNA sequence into a plant animal or microbe, the resulting ‘expressed’ string of amino acids in the protein will correspond to the recipe as provided in the DNA base sequence. This is true regardless of the species source, or the species recipient, as all living things use the same DNA language.

 

To ensure the inserted gene does what it is supposed to, various molecular, chemical and biological tests are conducted on the GM plant, animal or microbe to check that the inserted DNA sequence is intact and the expressed protein functional before the GMO is commercialized.  They are also extensively tested for safety and efficacy for several years prior to commercial release to ensure there are no unexpected or unusual results.

 

There are no known examples where a specific gene recipe (DNA sequence) was inserted into a cell and produced something other than the predicted protein.  What can—and does—go wrong is to have the inserted gene go unexpressed, or only partially expressed, such that the amount of protein is insufficient to prove commercial levels of the desired trait.  In those cases, the GMO is destroyed as soon as it is detected, usually in very early testing, and in any case long before commercialization.”

 

You might also be interested in a response to a similar question from Dave Kovalic, Regulatory New Technology Lead from Monsanto, in which he addresses genome insertion, an excerpt is below:

 

“…Researchers use a number of characterization techniques to understand and locate exactly where the gene was inserted, and subsequently select only those plants with the best insertion points (optimal molecular profiles) for further work.

 

Unlike new crop varieties developed by other breeding methods, researchers fully characterize GM crops at the molecular level. Following is a more technical description:

  • For any given “event” (events are the individual products of transformation), researchers will assess the number of distinct insertion sites resulting from transformation using a variety of molecular biology techniques (e.g. genome sequencing, PCR based methods or Southern blotting). We typically consider the events that contain a single insert as optimal, with others being identified and eliminated from further work.
  • For any insertion site, researchers can determine the DNA inserted and its location in the genome by sequencing, which involves determining the precise genetic sequence of A, C, G and Ts, in an event’s genomic DNA. By examining the inserted DNA, we can confirm that the insertion occurred as intended. Also by examining the genome sequence surrounding or “flanking” the insertion site, we can tell if any native coding or regulatory regions have been disrupted. Once again only events with the intended insertion and without disruption of native sequences are kept for further work.
  • Given this full characterization of the number and nature of insertions produced during transformation, we are able to identify and keep only the desirable events, with all others being excluded.
  • In addition, during the development of new varieties, a researcher will screen thousands of plants based on their appearance to make sure there are no apparent unintended effects. However, researchers do this for all new varieties whether they are being developed by GM technologies or through conventional breeding techniques.

Last, I’d mention that even though researchers fully characterize GMOs down to the level of their DNA, similar types of insertions have happened in nature many, many times over the history of cultivation and consumption of crops. Those resulting crops have remained safe for consumption in the context of all of these changes.”

 

If you have any additional questions, please ask.

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