CRISPR ACTIVITY

Q: What is “CRISPR”?
CRISPr is basically a type of immune system discovered in bacteria, and scientists can
adapt its components into a biotechnology tool for editing DNA. it can correct genetic
defects, treat and prevent the spread of diseases and improve crops, etc.
Where do CRISPRs come from?
CRISPR technology comes from the immune system of bacteria and archaea. These
organisms use CRISPR-derived RNA and various Cas proteins, including Cas9, to foil
attacks by viruses and other foreign bodies.
How does the system work?
First, the scientists introduce the Cas9-guide RNA complex into a cell, where it associates
and dissociates with the DNA.Cas9 will recognize and also bind to a three-nucleotide
sequence motif that is called PAM, that is abundant throughout the genome.
The scientists will synthesize the guise RNA to obtain a 20-nucleotide sequence that will
match a particular sequence in a cell's DNA that they want to target. When the guide RNA is
added to Cas9, it will guide it to this target sequence. This can be nearly any sequence as
long it is near a PAM motif. It can be part of a genes coding region that scientists want to
change.
When it binds to a PAM motif, Cas9 will unwind the DNA double helix. If the DNA at that
location perfectly matches a 20-nucleotide sequence within the guide RNA, the DNA and
matching RNA will bind by complementary base pairing.
If the sequence of the unpaired DNA strand is not an exact match to the 20-nucleotide
sequence within the guide RNA, Cas9 disengages from the DNA, which zips back up into a
double helix. If the sequences are a perfect match, the guide RNA base pairs with the
complementary DNA sequence, forming a DNA-RNA helix. This binding event activates
Cas9’s nuclease. It makes specific cuts in the DNA at a position three nucleotides upstream
from the PAM site. Two active sites on the nuclease domain of Cas9 generate the cuts and
cleave both strands of the DNA double helix, resulting in a double-stranded DNA break.
Cells contain enzymes that repair double-stranded DNA breaks. The repair process is
naturally error-prone and will lead to mutations that may inactivate a gene. Cleaving DNA at
a precise location is one of many applications of the CRISPR-Cas9 technology.

How does CRISPR-Cas9 compare to other genome editing tools?

Crispr-Cas9 can be compared to other editing tools by many things. First of all we can say
that CRISPrS do not need to be paired with separate cleaving enzymes like other tools do,
since they are capable of cutting DNA strands by itself. Also, they can be matched with
gRNA sequences that are designed to lead them to its DNA targets, and there about tens of
thousand of these sequences that have been created and available for research. Also it can
be used to target multiple genes simultaneously, compared to other gene editing tools.

How does CRISPR-Cpf1 differ from CRISPR-Cas9?

CRISPR-Cpf1 differs from CRISPR-Cas9 in different ways. Starting with its natural form,
since Cas9 DNA cutting enzyme forms a complex with tho small RNAs, required for the
cutting activity. On the other hand, Cpf1 is simpler since it only requires an only single RNA.
Its enzyme is smaller than the Cas9, so it benefits the delivery into cells and tissues.

Also, Cas9 complex cuts DNA, both strand at the same place, leaving blunt ends that are
prone to mutations since they are joined. With the Cpf1 complex, this will cut the two strand
offset, meaning that it will leave short overhangs on the exposed ends. This will help with
making a precise insertion, making it accurately.

The cpf1 cuts faraway from its recognition site, so if the targeted gene becomes mutated at
the cut site, it also can be re-cut, allowing opportunities to correct and edit.

Also the Cpf1 provides flexibility when choosing targeted sites. This compared to Cas9, can
recognize very different PAM sequences, being an advantage when targeting.

What other scientific uses might CRISPR have beyond genome editing?
CRISPR helps scientists in research creating cell and animal models very quickly, so they
can accelerate its investigation into diseases. It is nowadays being used as a rapid
diagnostic.

Propose two or three specific measures to help policy keep pace with CRISPR technology.
In explaining these measures, include details of the CRISPR-Cas9 technology, its
applications, and implications for dual use.
CRISPR-Cas9 should only be used to enhance and improve the conditions of living beings.
By this I mean, it must be used to help to fight diseases, improve the diagnosis, generate
more crops, help with the conservation of nature and the environment. It cannot be used for
individual benefits such as genetically modifying the phenotype for aesthetic.
There must be a measure of who can use CRISPR-cas9. I think that not everyone can use it
since it could be used for individual benefits and not to help the development of our society.
I believe that the scientific society should be the one that controls and uses CRISPR-Cas9
because they are the ones trained in the area, they know how to use it and in turn are the
only ones who can help to regulate the errors that may arise.
I think that a specific measure when using CRISPR-Cas9 is to create ethical and moral
regulations that must be used in CRISPR. These regulations must be approved and created
by a society with different points of view where the measurements, the limits and the
possible consequences it may cause are evaluated. With this, when evaluating all the
factors, the scientist must comply with the regulations stipulated to the letter, so that there is
a control of the use.
To what extent should dual-use technology be private, restricted, or prohibited? On a
continuous scale from total transparency to prohibition, choose and define a position. In
defending that position, use and explain details of the CRISPR-Cas9 technology, its
applications, and implications for dual use.
First of all I believe that dual-use technology should not be private. This is because if the
dual-use of technology is private, only certain people will have access to it, which will be
unfair to people who cannot acquire it. That is, only people with economic capacity will be
able to make use of this and modify their genome in this case with CRISPR-Cas9 for their
benefit. In my opinion, dual-use technology should be managed by the government in the
company of the scientific society where an ethical and moral law is applied. In these
regulations it must be determined how the CRISPR will be used and who are the ones who
can use it based on their condition.
On the other hand, I think that the use of CRISPR-Cas9 should be controlled, restricted to
cases in which health is impaired. That is, to cure diseases, diagnose, help in the growth of
crops, combat those hereditary diseases e.t.c. I think that it should be restricted only to the
use of health and factors that benefit us as a society. I think this is tied to allowing the
dual-use of technology to be controlled by the private or public sector. If it is misused, and if
the CRISPR-Cas9 is in the hands of the private sector, probably it will not only be limited to
benefiting health but also extends to wanting to modify those physical and superficial
attributes, such as changing our phenotype because of aesthetics. This would not only be
ethical and moral, since this technology is used in a way that does NOT help us as society,
but also because it is in the hands of the private sector as mentioned above, it will not be
affordable for everyone. The privileged sector would be the only one acquiring this quality.
I believe that CRISPR should not be prohibited since it has many benefits that will help us
improve our world. CRISPR is capable of modifying genes in our favor where we can fight
diseases, helping the preservation and comfort of living beings.