AUGUST 9, 2015

BLOG, SPECIAL EDITION ON GMOS

HOW TO MAKE A GMO

by Chelsea Powell
figures by Anna Maurer

Summary: Genetically modified organisms (GMOs) are

organisms that have been altered using genetic engineering
methods. Although genetic engineering is a common and
essential practice in biotechnology, its specific use in crops is
controversial. The key steps involved in genetic engineering
are identifying a trait of interest, isolating that trait, inserting
that trait into a desired organism, and then propagating that
organism. Methods for genetic manipulation have rapidly
improved over the last century from simple selective breeding,
to inserting genes from one organism into another, to more
recent methods of directly editing the genome.

What qualifies as a GMO?

A common misconception is that any animal or plant
considered to be outside the realm of our reference for
“natural” is a GMO. Images of abnormally large cows and
tomatoes come to mind. However, the scientific community
and the U.S. Food and Drug Administration (FDA) use a
stricter definition for a GMO: an animal or plant that has been
created through genetic engineering [1]. Genetic engineering
is a term used to describe biotechnological methods used by
scientists to directly manipulate an organism’s genome. Under
this definition GMOs do not include plants or animals made by
selective breeding, or animals modified by being given
hormone supplements or antibiotics. In fact, we do not
currently eat any meat products considered to be GMOs,
although farm animals may be fed a genetically modified crop
[2].
The main goal of the majority of genetic engineering
performed on food is to increase crop yield and/or to improve
the nutrient value in animal feed. No genetically engineered
crops on the market in the United States have been modified
to be unusually large (Table 1). Pictures of extremely large
vegetables used to support the “Franken-food” image of
GMOs are probably not GMOs at all; an unusually large
vegetable would more likely be created through less
controversial methods of selective breeding or nutrient
supplements, not genetic engineering.

The process of genetic engineering

Genetic engineering is widely used in biological research.
Mouse models are engineered for biomedical studies, bacteria
are engineered to produce medications such as insulin, and
crops are engineered for agriculture. All of these products of
genetic engineering were created using the same basic steps:
identifying a trait of interest, isolating that genetic trait,
inserting that trait into the genome of a desired organism, and
then growing the engineered organism (Figure 1). These steps
are explained in detail below, using examples from Monsanto
as the details of their technologies are publicly available.
Other major companies such as Syngenta, BASF, Dow,
Bayer, and Du Pont use similar methods, as outlined in brief
on their respective websites [3, 4, 5, 6, 7].
Step 1: Identify a trait of interest
In order to identify a desirable new trait scientists most often
look to nature. Successful discovery of a new genetic trait of
interest is often a combination of critical thinking and luck. For
example, if researchers are searching for a trait that would
allow a crop to survive in a specific environment, they would
look for organisms that naturally are able to survive in that
specific environment. Or if researchers are aiming to improve
the nutritional content of a crop, they would screen a list of
plants that they hypothesize produce a nutrient of interest.
An example of a trait currently in GMOs that was identified
through this combination of luck and critical thinking is
tolerance to the herbicide Roundup (see this article).
Monsanto created “Roundup Ready” plants after finding
bacteria growing near a Roundup factory that contained a
gene that allowed them to survive in the presence of the
herbicide [8]. Although it is not on the market in the United
States, Syngenta has designed Golden Rice with an increased
amount of pro-vitamin A, which the human body may turn into
the vitamin A (see this article). Researchers at Syngenta
identified the gene sequence that produces pro-vitamin A and
compiled a list of plants to screen with that sequence [9]. With
a little luck, there was a plant in nature, maize, that contained
a gene that would make Golden Rice produce pro-vitamin A at
a level that could meet the nutritional needs of vitamin A
deficient communities.

Step 2: Isolate the genetic trait of interest

Comparative analysis is used to decode what part of an
organism’s genetic makeup contains the trait of interest. The
genomes of plants with the trait are compared to genomes in
the same species without the trait, with the goal of identifying
genes present only in the former [8]. The genomes of different
species with the same trait may also be compared in order to
identify a gene, as was the case while developing Golden Rice
[9]. If there is no database of genetic information for
comparison, scientists will purposefully delete, or “knock out,”
parts of the genome of interest until the desired trait is lost,
thereby identifying the genes that lead to the trait.

In order to expedite this process, Monsanto has developed

and patented a method known as seed chipping [8]. Through
this method Monsanto shaves off parts of seeds for high-
throughput genetic sequencing while leaving the rest of the
seeds viable for planting. This creates a genetic database for
plants before they are even grown, where a barcode system is
used to match plants to their genotypes. Researchers may
then use this database to identify new traits of interest as well
as to optimize the desirable traits in a crop by selecting for the
best genotypes based on plant phenotypes.

Step 3: Insert the desired genetic trait into a new genome

Altering the genome of plant seeds is difficult due to their rigid
structure. Many biotech companies use “gene guns” that shoot
metal particles coated with DNA into plant tissue with a .22-
caliber charge [8]. Monsanto no longer uses gene guns, but
instead takes advantage of bacteria, called Agrobacterium
tumefaciens, that naturally invade seeds and alter plants by
inserting pieces of their own DNA into a plant’s genome.

In biotechnology research it is common to genetically engineer

bacteria to produce a desired protein. This is done by using
enzymes to cut and paste a DNA strand of interest into a
plasmid, which is a small, circular molecule of DNA [10].
Bacteria are then shocked using heat or electricity so that the
cells accept the engineered plasmid. By modifying A.
tumefaciens, which is easier to modify than plant seeds
themselves, researchers may use the bacteria’s naturally
invasive behavior as a Trojan horse for inserting desirable
traits into a crop’s genome.

Step 4: Growing the GMO

After a genetic trait has been successfully inserted into an
organism’s genome, the modified organism must then be able
to grow and replicate with its newly engineered genome. First,
the genotype of the organisms must be checked so that
researchers are only propagating organisms in which the
genome was modified correctly.

Biotech companies invest large sums into keeping these

plants alive and reproducing once they have been
successfully created. The companies use special climate-
controlled growth chambers, and biologists often check on the
plants by hand to make sure that they are growing as
expected [8].
During this process biotech companies will use automated
machines, such as Monsanto’s GenV planter, in order to track
plants and calculate optimal seeding and growth conditions to
create the best possible yields. GMO seeds often come with
instructions on spacing and nutrition that result from these
studies.

Future directions for the creation of

GMOs
Humans’ ability to modify crops for improved yields and
nutrients in a given environment is a keystone of agriculture.
The technological advancement from selective breeding to
genetic engineering has opened up a large realm of
possibilities for the future of our food. As techniques for
genetic engineering, such as new RNAi- and nuclease-based
technologies that allow for direct modification of the genome
(see this article and this article), steadily improve, our ability to
create new GMOs will also grow [11]. As our scientific
capabilities expand it is essential that we discuss the ethics
and ideals surrounding GMOs so that we may effectively and
safely use this technology in a way that is acceptable to the
public.