CRISPR Extended Definition

Charmayne Zablan

ENG 363 Scientific Writing

 WHAT IS CRISPR
A clustered, regularly interspaced,
short, palindromic repeat (better known as
CRISPR) is an immune system used by
bacteria to fight off viruses. Researchers
have taken notice of a bacterial DNA that is:
 repetitive in sequence;
 often followed by the same
sequence in reverse and;
 mixed with fewer spacer DNA in
between,
hence the name of the system—regularly
interspaced, palindromic (same sequence in
reverse), and repeat (i.e. Figure 1) (Pennisi,
2013).
For a while, the oddly coincidental
palindromic repeats went unnoticed until
scientists realized how bacteria used them as
an immune strategy (Pennisi, 2013). Shortly
after its discovery, scientists from around
the world realized CRISPR can be used as a
gene editing tool. This awareness caused a
rapid uproar in the scientific community
(Baker, 2014).
Figure 1. CRISPR at work in a prokaryotic cell. The
“CRISPR locus” in the figure shows how a CRISPR
looks with its DNA and spacers. Reprinted from
CRISPR Systems, Doudna Labs, 2020, retrieved from
http://doudnalab.org/research_areas/crispr-systems/
HOW CRISPR AND CRISPR-CAS
SYSTEMS WORK
In Bacterial Cells
CRISPR is used in a bacterial
immune system to fight of viruses. The
CRISPR system recognizes foreign DNA
(that has been inserted by a virus) in the cell
and makes a cut into its own DNA. Then
CRISPR inserts the foreign DNA into the
bacterial DNA, so it is recognized, and the
bacteria could battle off future similar
viruses (Terns & Terns, 2014). Figure 1
shows how CRISPR works in a bacterial cell
setting.
CRISPR-Cas Systems
The CRISPR-Cas9 system is one of
the most common CRISPR-Cas systems
(Cas: CRISPR associated) used for gene-
editing in human DNA. This is because it is
simple to harness and has become very fine-
tuned (Pennisi, 2013). The CRISPR-Cas9
system includes CRISPR, the Cas9 protein,
and [usually] a guide RNA (Knott &
Doudna, 2018).
Figure 2. This infographic breaks down the CRISPR-
Cas9 system inserting DNA into 4 basic steps.
Reprinted from MRSBulletin, Retrieved February
2020, from
https://www.cambridge.org/core/journals/mrs-
bulletin/news/crispr-implications-for-materials-
science. Copyright 2016 by Philip Ball. Reprinted
with permission.
The guide RNA creates a complex with the
Cas9 protein which is able to locate (through
the guide RNA) and cut a targeted DNA
(through the Cas9 protein). (Baker, 2014;
Knott & Doudna, 2018; Pennisi, 2013).
HOW CRISPR IS APPLIED IN THE
SCIENCES
The CRISPR-Cas systems are such a
fast-paced research topic that the list of
organisms (plant and animal) worked on
with the technology is large and ever-
growing (Baker, 2014).
There are three different ways that
the CRISPR-Cas system can disrupt genes:
1. Cutting and inserting new DNA
sequences,
2. Repressing, or turning off, genes,
and
3. Activating, or turning on, genes.
(i.e. Figure 2) (Pennisi, 2013).
 CRISPR Applied Speed and Ease

One example of the use of Unlike the two other gene-editing

technology is the correction of cataract- tools, CRISPR-Cas systems do not rely on

causing genes (in mice), performed by the use of a protein, giving the advantage of

researchers from the Shanghai Institutes for speed and ease over its predecessors (Baker,

Biological Sciences (Baker, 2014). 2014). Where it would take ZFNs or

Another example of CRISPR TALENs up to months to harness, CRISPR

progression is that it has been used to could do the same work in about a week,

suppress genes in wheat that allow mildew because you do not have to create a

resistance (Pennisi, 2013). The list completely new protein like you would with

continues, but with caution due to the ZFNs and TALENs (Baker, 2014; Pennisi,

technical issues and ethical concerns that 2013).

can be found in “The Issues with CRISPR Adding onto speed, the CRISPR-

Technologies”. Cas9 system was used to show that it can cut

multiple genes at the same time. Results

WHY CRISPR IS THE FAVORABLE showed that the system was successful in up
CHOICE OF GENE-EDITING TOOL
to [a reported] 5 genes at once (Pennisi,
There are two other common gene-
2013).
editing tools that were widely used before
Accuracy
the discovery of CRISPR-Cas systems. One
CRISPR-Cas systems have also
of those tools is Zinc Finger Nucleases
shown more accuracy than other gene-
(ZFNs) and the other Transcription
editing tools through its efficiency at
Activator-like Effector Nucleases
locating and cutting correctly targeted genes
(TALENs).
(Pennisi, 2013). In that same study showing
efficiency, Church was able to synthesize
thousands of RNA sequences to work with
CRISPR-Cas9 and showed that 90% of
human genes can be easily targeted and
manipulated successfully (Pennisi, 2013).
Low Costs
Another reason CRISPR might be a
favorable choice is that is low in cost
compared to other gene-editing tools. The
main reason it is cheaper is because it takes
so much less time to function. The software
relative to CRISPR-Cas systems are also
low cost; ranging from free to as little as $65
you can learn to design gRNA (guide RNA)
or make a personal CRISPR system
(Pennisi, 2013; Samy, 2017).
Figure 3. This graph shows the price comparison in
the utilization of TALENS vs. CRISPR technology.
CRISPR technology shows to be 3-6 times cheaper
than TALENS technology, depending on what type
of gene manipulation occurs. Reprinted from Gene
Editing Could Modify and Cure Disease: CRISPR vs.
TALENs, Samy, 2017, from https://ark-
invest.com/research/crispr-vs-talens
THE ISSUES WITH CRISPR
TECHNOLOGIES
Although CRISPR shows a great
deal of advantages over other gene-editing
tools, there are still some technical and
ethical concerns with the technology.
Technical Concerns
Even though it has shown greater
accuracy than ZFNs and TALENs, CRISPR
is still not completely accurate at targeting
genes. In some of J. Keith Joung’s work,
inaccuracy is shown through the alteration
of DNA that is similar to the targeted DNA
(Pennisi, 2013). Scientsts believe that the
system needs full fine tuning before it’s
considered truly safe, even with assuring
results (Pennisi, 2013). Furthermore,
CRISPR editing is permanent so once a cut
is made, the unknown consequences from
making an inaccurate cut are permanent as ruling out diseases in babies even before

well (Baker, 2014; Pennisi, 2013). birth, but it can pose a large threat of

Ethical Concerns eugenics, controlled human breeding based

Ethical concerns are also being off of “desirable” traits (Brokowski & Adli,

raised with the use of CRISPR, specifically 2019). CRISPR costs are low for

in the use of humans. Human embryos are functioning in the lab (the costs mentioned

up to debate with the use of CRISPR-Cas above are just the software costs and do not

systems, largely due to the controversy on include all costs including actually going

whether or not an embryo is already a into DNA to edit, etc.) so anyone that could

human holding the power to make its own consider gene-editing would be those with

bodily decisions (Brokowski & Adli, 2019). abundant monetary resources. This is a

CRISPR-Cas use in “regular” adult concern for society as money and beauty

humans has only recently been considered as come into play, because wealth can

it is a touchy subject because we do not truly determine favorable features, challenging

know the long-term outcomes and/or the eugenics and possibly evolution.

permanent side effects of the technologies

(Baker, 2014; Pennisi, 2013). It is also

unknown how CRISPR-Cas systems will

affect someone’s germline and eventually

their offspring.

The largest ethical concern with

CRISPR is the rise of “designer babies”.

CRISPR-Cas systems help largely with

 Works Cited

Baker, M. (2014). Gene editing at CRISPR speed. Nature Biotechnology, 32(4), 309+. Retrieved

from https://link-gale-com.lib-proxy.fullerton.edu/apps/doc/A369762573/OVIC?

u=csuf_main&sid=OVIC&xid=a7a818b9

Ball, P. (2016). CRIPSR: Implications for Materials Science. MRSBulletin.

https://www.cambridge.org/core/journals/mrs-bulletin/news/crispr-implications-for-

materials-science

Brokowski, C., & Adli, M. (2019). CRISPR Ethics: Moral Considerations for Applications of a

Powerful Tool. Journal of Molecular Biol

ogy, 431(1), 88-101.

Doudna Lab. (2020) CRISPR Systems. http://doudnalab.org/research_areas/crispr-systems/

Knott, G., Doudna, A. (2018) CRISPR-Cas guides the future of genetic engineering. Science,

361 (6405), 866-869. Retrieved February 10, 2020, from https://science-sciencemag-

org.lib-proxy.fullerton.edu/content/361/6405/866

Pennisi, E. (2013). The CRISPR Craze. Science, 341(6148), new series, 833-836. Retrieved

February 10, 2020, from www.jstor.org/stable/23491231

Samy, M. (2017). Gene-Editing Could Modify and Cure Disease: CRISPR vs. TALENs. Ark

Invest. https://ark-invest.com/research/crispr-vs-talens

Terns, Rebecca M, & Terns, Michael P. (2014). CRISPR-based technologies: Prokaryotic

defense weapons repurposed. Trends in Genetics, 30(3), 111-118. Retrieved from

https://www-sciencedirect-com.lib-

proxy.fullerton.edu/science/article/pii/S0168952514000146
Post write: The first definition strategy I used was parenthetical definitions, I used them

throughout a lot of the extended definition because my word choice required a lot of background

information. I also used examples in my extended definition which can be found under How

CRISPR is applied in the sciences. Partition was a large part of my definition because I felt that it

needed that type of organization to understand easily. Another strategy I used was negation,

which can be found in Why CRISPR is the favorable choice of gene-editing tool.

Acknowledgements: I would like to acknowledge Matthew Kim. His comment about language

made me realize that I was using language that a non-general audience can understand. I looked

back at the word usage and did my best to use parenthetical definitions for lesser known terms.

Source Evaluation Grid:

Criteria #2: CRISPR-Cas guides the future of genetic

#1: The CRISPR Craze engineering
relevant in terms of
Relevance: (e.g., background info, how it is
tangentially related vs. used/performed, examples of
essential to your what it's been used on, things
subject) that still need consideration.
One of the most relevant relevant for summarization of basic mechanisms of
sources I used. CRISPR-Cas systems

Timeliness: (e.g., old,

sort of recent,
happening now)

sort of recent (2013) recent (2018)

Authority: (e.g., expert

or amateur audience,
biased or balanced)
for an amateur audience for a more informed audience on the matter, balanced
(from a textbook) as it is a scientific article and not from a blog type
Evidence: (e.g., plentiful
or limited, types,
sources identified or
not) a good amount of sources,
direct quotes from
researchers and paraphrasing plentiful sources, identified using the number system

#3: Gene editing at

CRISPR speed #4: CRISPR Systems

relevant in terms of
background info,
consequences, other gene-
editing tools. Another very
relevant source used. relevant, easy how-to read

most recent of all sources

sort of recent (2014) (2020)

for a more informed audience, steered towards an amateur

very balanced because it is a audience, more of a pleasing
peer-reviewed scientific website to look at (although
article had good information)

good authority, plentiful limited sources, not even

resources really listed at all in website