Magistri 1

Lee Magistri
ENGL 1001-23
Mr. Dorhout
11/14/17
The Case for CRISPR/Cas9

Genetic engineering has been a hot topic issue since 1973. The latest and greatest genetic

engineering technology, CRISPR/Cas9, has not been able to avoid skepticism. CRISPR/Cas 9 is

a relatively new technology to edit genomes. CRISPR/Cas9, which is short for Clustered

Regularly Interspaced Short Palindromic Repeats, are specific stretches of DNA. These stretches

contain two parts: nucleotide repeats and spacers. The nucleotide repeats are the building blocks

of DNA. The spacers are strains of DNA connecting the nucleotide repeats. Researchers

discovered that within bacteria, the spacers contained information of previous viruses. Bacteria

used the memory of bacteria to resist and attack any time that virus came back. Cas9 is a protein

that acts on the memory to cut up the foreign invader. Researchers discovered that they can use

this technology to remove and replace DNA fragments, which offers researchers a simple and

effective way to alter DNA sequences and modify genes. Cas9 has already been used for

genome editing in a wide variety of organisms and cell types ranging from bacteria, yeast,

zebrafish, roundworm, fruit fly, crop plants, mouse, goat, monkey, to human cell lines...

(Rajendran et al. 264).

CRISPR/Cas9 has been a highly debated technology due to its ability for humans to alter

and rewrite the foundation of any living organism. With new research, CRISPR/Cas9 offers

humans to jump medical and agriculture hurdles. It allows the correction of gene based diseases

and prevention of vector borne diseases. It also has huge implications on the food science
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industry by increasing agricultural yield as well as identifying harmful bacteria.With the

multitude of benefits this technology provides for society, it comes with some moral dilemmas.

This is a powerful new tool and is one that needs a global organization to provide standards and

protocols.

Many of the early applications of CRISPR-Cas systems actually arose from food

science-driven research during characterization of industrial starter culture bacteria for

improving milk fermentation processes, (Selle and Barrangou R2367). The use of

CRISPR/Cas9 technology was recognized in the food industry due to its ability to revolutionize

the quality and quantity of food. CRISPR/Cas9 can be used as a quick way to identify bacteria. It

is also a great tool in genome editing and as a gene drive passing on codes for future crops and

livestock.

Identifying bacteria in food is crucial for human consumption, especially during

foodborne outbreaks. Humans have been able to identify bacteria in food for quite some time.

Some of the ways we identified bacteria in food were genome sequencing, pulsed field gel

electrophoresis, and repetitive-element PCR-based genotyping. Although we have been using

these methods for years, they are time consuming, complex, and expensive. What CRISPR/Cas9

allows scientists is a quick, efficient, and cheap tool for them to identify bacteria strains with

high resolution. To date, CRISPR-based typing schemes have been effectively employed in

foodborne pathogens such as Salmonella and Escherichia coli, industrial fermentation starter

cultures such as S. thermophilus, probiotics such as Lactobacillus casei, and spoilage organisms

such as Lactobacillus buchneri, illustrating the broad potential of CRISPR-based genotyping

across the bacterial spectrum (Selle and Barrangou R2369).

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Cas9 can be harnessed to target individual genes or multiplexed for targeting several

gene family members (Rajendran et al., 267). This genome editing is a crucial step in

maintaining livestock and in the reproducibility of crops. CRISPR/Cas9 has already been applied

to tomatoes, corn, rice, tobacco and yeast. It has been proven to improve growth even during

drought conditions, low fertilized conditions, and improve nutritional levels in foods.

CRISPR/Cas9 can also edit genetics, eliminating diseases and viruses. If scientest can eliminate

diseases and viruses just through manipulating the DNA, there would be no need to pump

excessive vaccinations and antibiotics into the animals. It has also been shown to possibly

remove some allergenic genes which would allow for individuals to consume nutritional food

that otherwise would have been deadly. Biofortification of milk through the knockout of

allergenic gene such as b-lactoglobulin in genome-engineered dairy animals might be possible in

the future, (Rajendran et al., 270). Not only can you provide these benefits to livestock and

crops, but gene drives pass on changes of DNA from parent to offspring. This allows future

crops and livestock to be resistant to diseases, have higher nutritional value, and grow in

unfavorable conditions. Increasing the quality and quantity of agriculture can be a useful tool in

combating food shortages. Providing an abundance of food means more food can go to the

hungry and impoverished.

Besides all the possibilities CRISPR/Cas9 provides for agriculture, some may argue that

CRISPR/Cas9 has more potential as a therapeutic tool in the healthcare field. As a powerful

genome editing tool, there has been research that CRISPR/Cas9 can eliminate insects as vectors.

CRISP/Cas9R has been used on domestic animals to help produce biological medical materials,

such as organs and cells. Furthermore, it has also been used to study the treatment of human

diseases. The benefits of using CRISPR/Cas9 in the biomedical field seem endless.
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Malaria is a disease caused by a parasite. It is passed into humans through mosquitos. In

2015, it is estimated that malaria killed over 429,000 people. Researchers have discovered two

ways CRISPR/Cas9 can stop the transmission of malaria, potentially saving millions of lives.

Human malaria requires development of an endonuclease-based gene drive system that

interferes with the ability of A. gambiae mosquitoes to transmit the disease. This could be

achieved either by blocking parasite development or by reducing the reproductive capability of

the insect vector. (Hammond et al., 79). Furthermore, CRISPR/Cas9 has a germline genome

editing ability. Gene drive systems allow for a specific trait to have a higher possibility to be

passed down to offspring. The rate of the specific trait to be passed down increases with each

generation. So the two methods used to halt the transmission of malaria would not be a

temporary solution. But, it would be permanent affectively changing the DNA of the mosquito

forever. This new technology opens the door to eradicating all vector borne diseases.

There are currently over 120,000 people in the United States on a wait list for an organ

transplant. The average time people will wait to receive a transplant is from two to four years. If

patients need an organ, most likely they do not have two to four years to receive a transplant.

Researchers have been considering xenotransplantation as a method to stop this donor organ

shortage. Xenotransplantation is transplanting organs or cells from an animal to humans. Two

complications that derive from this transplant are passing diseases from animals to humans and

patients immune system rejecting the tissue. The main animal that humans would use to develop

organs from are pigs. Pigs can be bred in a pathogenic environment and are domesticated,

making them a suitable choice. The problem with transplanting pig organs to humans is that

under stress, pigs release porcine endogenous retrovirus (PERV). PERV can infect humans,

which is a hurdle researchers must overcome before preforming xenotransplantation. Using an

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approach called the CRISPR (clustered regularly interspaced short palindromic repeats)Cas9

system, which targets a specific DNA sequence for disruption, they genetically engineered a one-

step inactivation of more than 60 copies of PERV, thereby reducing the infectious risk from

PERVs by three orders of magnitude.. (Phimister and Salomon, 1089). There are a few other

complications that come along xenotransplantation. CRISPR/Cas9 allows one less hurdle that

researches must jump to end this organ donor shortage.

CRISPR/Cas9 can eradicate gene based diseases such as Sickle Cell Anemia,

Huntingtons disease and many more. Gene based diseases are passed down through DNA. Gene

editing has never been an affordable and simple procedure to perform. CRISPR/Cas9 allows

researchers the ability to access embryos and change DNA. CRISPR/Cas9 has also been shown

to eliminate a gene that is necessary to for HIV to be integrated into a host. Khalili lab has

delivered the CRISPR/Cas9 system via a tail-vein injection to target a HIV gene which is crucial

for the integration of viral DNA into the host genome.67 These treated animals demonstrated a

reduced expression of HIV gene in multiple tissue organs, implicating a reduction in viral

infectivity produced by CRISPR editing in vivo. (Cai et al. 248). Affectively eliminating the

trait of gene based diseases to be passed down. With the germline ability, it is possible to

eliminate gene based diseases forever.

Genome editing is a bioethical dilemma that has been discussed for decades and one that

will continue to be discussed in the future. Since CRISPR/Cas9s ease of use, the debate over

genome editing has been rekindled. CRISPR/Cas9 has the potential for designer babies to affect

future generations. It also opens the door to Frankenstein creations, potentially pushing humans

to the next step in evolution. Leaving the world with a necessary discussion on what limits will

be implemented for this technology and who should make the standards.
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People are afraid that CRISPR/Cas9 allows designer babies to not only affect your child

but future generations. If thirty percent of Americans want their child to have green eyes and red

hair, CRISPR/Cas9 has the potential to strengthen these traits to be passed down. Not only would

the first generation have a greater chance of expressing these traits, but their children would have

a greater chance to express these traits. Through time this will eliminate all other traits besides

the desired ones. It is not ethical for people to decide if others should look short, tall, skinny fat,

red hair, green eyes etc. Therefore, killing off certain physical traits and creating a less diverse

world is not a decision that can be made for future generations.

Since CRISPR/Cas9 is simpler and cheaper than its predecessors, using its technology

outside of government regulations is a discussion that must be had. Genome editing has been

used before to alter animals, most notorious example is Eduardo Kac's creation of (or order to a

genetics laboratory for) a green fluorescent protein rabbit created by injecting GFP protein into

fertilized rabbit eggs and then breeding any rabbits that had taken up the gene (Charo and

Greeley, 13). How far will people push the boundaries with such creations and what if people

create unforeseeable evils? Synthego, a leading provider of genome engineering solutions,

today announces the world's first synthetic single guide RNA (sgRNA) CRISPR genome editing

kit, (BioMedReports 1). This kit will be sold for 295$, for personal use, and with discounts for

academic research. This is a grossly irresponsible product giving anyone the chance to create

monsters.

With the possibility of creating monstersand killing off races, it is obvious there needs to

standards with CRISPR/Cas9. The world needs to have a discussion on who should make

standards for this technology. If each country makes their own standards for CRISPR/Cas9, it

can lead to some unleveled playing grounds. The United States and Russia have been rivals for
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quite some time, with each country trying to get one step ahead of each other when it comes to

military technology. Imagine America decides they are going to use CRISPR/Cas9 not only for

food science or medical benefits but we also use it for enhancement. They start implementing

CRISPR/CAS9 in every child. Passing on traits to make them smarter, taller, and stronger. After

generations this can turn humans into super soldiers. Now imagine Russia only used

CRISPR/Cas9 for medical and food advancement. They would be at a clear disadvantage if there

were to be another world war. Besides that, after so many generations down the road there would

be a clear separation between people that used CRISPR/Cas9 for enhancement and people that

did not. This separation could be a kickstart in the next step in the evolution process, leaving the

less advanced people behind.

Therefore, a global regulation on CRISPR/Cas9 is a necessity to ensure that scientist do

not make any detrimental decisions without a global discussion. Although scientist have faced

some bioethical dilemmas, they should push on with research. This tool has the possibility to

eradicate diseases and end the organ donor shortage. It provides the possibility to end world

hunger and catch food borne outbreaks before they cause to much damage. It is not the

technology scientist should fear, but it is what they do with it.

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References

Phimister, Elizabeth G., and Daniel R. Salomon. "A CRISPR Way to Block PERVs --
Engineering Organs for Transplantation." The New England Journal of Medicine, vol.
374, no. 11, 2016, pp. 1089-1091, Research Library,
https://search.proquest.com/docview/1774194815?accountid=2909.
Hammond, Andrew, et al. "A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction
in the Malaria Mosquito Vector Anopheles Gambiae." Nature Biotechnology, vol. 3
4, no. 1, 2016, pp. 78-83, Agricultural & Environmental Science Database; ProQuest
Technology Collection; Research Library; Technology Collection,
https://search.proquest.com/docview/1754588758?accountid=2909,
doi:http://dx.doi.org/10.1038/nbt.3439.

Rajendran, Subin Raj Cheri Kunnumal, et al. "CRISPR-Cas9 Based Genome Engineering:
Opportunities in Agri-Food-Nutrition and Healthcare." OMICS: A Journal of Integrative
Biology, vol. 19, no. 5, 2015, pp. 261-275.

BioMedReports: Synthego Announces World's First Synthetic Single Guide RNA Kit for
CRISPR Genome Engineering. Newstex, Chatham, 2016, ABI/INFORM Collection,
https://search.proquest.com/docview/1821165656?accountid=2909.

Charo, R. A., and Henry T. Greely. "CRISPR Critters and CRISPR Cracks." The American
Journal of Bioethics, vol. 15, no. 12, 2015, pp. 11-17.

Selle, Kurt, and Rodolphe Barrangou. "CRISPR-Based Technologies and the Future of Food
Science: CRISPR Applications in Food Science." Journal of Food Science, vol. 80, no.
11, 2015, pp. R2367-R2372.

Cai, Liquan, et al. "CRISPR-Mediated Genome Editing and Human Diseases." Genes &
Diseases, vol. 3, no. 4, 2016, pp. 244-251.