GENE EDITING TECHNOLOGY

Ashok Karre

Information Technology

Bournemouth University

Bournemouth, United Kingdom

S5229648@bournemouth.ac.uk

Abstract—This report includes the different
technologies associated with the editing of the gene. At
the molecular level, alteration of the DNA of interest
can be executed with the implementation of different
gene editing technologies which includes CRISPR/Cas9
(Clustered regularly interspaced short palindromic
repeats which are associated with protein 9), TALEN
(transcription activator-like effectors nuclease), and
ZFN (zinc finger nuclease). These gene-editing tools are
frequently used in the field of molecular biology that
allows research and scientists to make changes in the
gene to produce different precaution and treatment for
different chronic health diseases. This report includes
the advantages and disadvantages associated with the
use of such gene-editing tools. With the help of such
tools and technologies, the genetic information of any
virus and DNA of interest can be studied and all the
relevant information has been provided in the latter
part of this report.
Keywords—CRISPR/Cas9, TALEN, ZFN
phenomenon is referred to as nuclei engineering and
through this process, the DNA can be modified and
can be further inserted into the gene of interest.[5]
However, targeted DNA can be induced to get
desirable results and such a process breaks the DNA
and later activates it through the help of DNA repair
mechanisms, and such parts of DNA are inserted into
site-specific areas in the genome of interest.
However, in the process of gene-editing technology,
the basic principle is related to the targeting of the
gene of interest which can be incorporated through
homologous recombination or the process of
conditional targeting. Moreover, the double-strand
breakage that occurs due to the cutting of the DNA of
interest can be repaired with the help of the double-
strand break repair mechanism which influences the
joining of the double-stranded DNA and allows in
further transcription and translation to produce
targeted proteins and mRNA.

II. BACKGROUND
I. INTRODUCTION
One of the frequently used gene editing
Gene-editing technology is a process of versatile
technologies is related to the CRISPR Cas9
technologies that has motivated researchers and
mechanism. However, other mechanisms are related
scientists to edit the genes to sequence genes
to the gene-editing process. Gene editing is also
according to the modified genome sequence. With
known as gene engineering because it provides the
the help of gene-editing technology, many
cutting and pasting of genes of interest to provide
researchers have initiated the modification of genes
desired results. With the help of this process, the gene
into the genome of those organisms that belong to a
of interest can be randomly inserted into the
broad spectrum. One of the frequent technologies
palindromic sequences to understand the mechanism
which are related to gene-editing technology is the
of the gene of interest. It helps in the modification of
CRISPR-Cas9 and TALEN. These two technologies
the DNA used in an organism to study the
are highly recommended and frequently used in the
consequences of the desired gene.[6] The process
editing of genes. CRISPR-Cas9 which is called
involves three major steps that include insertion,
Clustered regularly interspaced short palindromic
deletion, and modification of the gene of interest.
repeats which are associated with protein 9.[10] With
With the help of this process, researchers and
the help of such innovative technologies, researchers
scientists can understand the deadly diseases that
can develop new genomes that can be used in the
occur in human beings, and through the alteration of
treatment of different kinds of diseases especially
DNA in the targeted organism, researchers and
cancer. The process is executed by cutting the DNA
scientists can treat such diseases to understand the
sequence with specific enzymes and thus this
future scope and consequence. With the help of such

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technology, various kinds of medicines and vaccines
can be established to get various medicines that can
treat chronic disease. Gene-editing technology has
made the researcher believe that certain diseases like
HIV, cancer, and hemophilia can be treated which
can help to cure millions of patients who are
suffering from such diseases.
III. DIFFERENT TECHNIQUES OF GENE
EDITING
Various techniques are related to gene-
editing technology and through this process,
researchers and scientists can develop techniques.
The techniques that are frequently used are.
A. CRISPR/Cas9
Clustered regularly interspaced short
palindromic repeats (CRISPR/Cas9) which is one of
the latest trends in the field of gene-editing
technology. This process is one of the quickest
processes that help researchers to insert the gene of
interest into the genome of organisms.[4] This
process of gene editing has been derived from the
CRISPR family present in bacteria which allows the
bacteria to detect the virus and this array of the gene
allows the bacteria to get protected from the virus.
However, this mechanism is highly recommended to
analyze the purpose of the foreign organism or DNA
that occurs within the cells.[2] Microorganisms that
include such a genome detect the virus and this
process can be executed in vitro in the laboratories.
This system allows the organism to involve the
implementation of the nuclease of cas9 to produce a
site-aimed double-fiber DNA break with the help of a
tracrRNA molecule that functions as a pilot
molecule. The nuclease of the cas9 functions as a
DNA strand breaker that allows the breakage of
double-stranded DNA.
B. Transcription activator-like effector
nuclease (TALEN)
TALE is a kind of protein that works as a
bacterial effector and it allows for analyzing the
development of DNA binding domain and such
domains are formed without the incorporation of
nuclease. This technology focuses on the use of
engineered nuclease which is referred to those
artificial proteins that are made by the researchers to
obtain customized sequence-specific DNA binding
domains.[3] These DNA binding domains are
synthesized in such a way to obtain desirable results
which are fused to a specific nuclease which later
cleaves the DNA in those non-specific sequence
order. The targetable nucleases are used to generate a
double-strand break in the double-stranded DNA into
the targeted sites.[8] These double-stranded breaks
are referred to as DSB and the phenomenon of using
transcription activator-like effectors nuclease
(TALEN) is frequently used to generate therapeutic
agents for those diseases that are generated through
genetic disorder.
C. Zinc finger nuclease (ZFN)
Zinc fingers are referred to those DNA binding
domains which are generally identified by three base
pair of DNAs. Change in the residue which can be
either a minor change around the alpha helix present
in the domain can result in the change of the
specificity of the DNA binding site.[1] A huge
amount of zinc finger arrays are widely engineered
with the help of different gene-editing technology by
fusing the gene with the non-specific nuclease
domain which is associated with the Type II FokI
restriction enzyme to synthesize zinc finger nuclease
which can be used in the targeting of the genome to
alter the changes in the gene.
IV. APPLICATION
The application that is associated with the
application of gene-editing technology includes
various processes that can be implemented in
laboratories so that various diseases can be cured
with the implementation of gene-editing
technology.[13] One of the frequently used
applications of gene-editing technology includes
targeted gene mutation that can give rise to a
mutation in the desired site of interest by inserting or
deleting the DNA sequence of interest. Gene therapy
is another approach of gene-editing technology that
allows curing diseases like cancer and other immune-
related diseases. The rearrangement of chromosomes
can be executed with the help of gene-editing
technology and the study of stem cells can be
executed to study the possible research and future
scopes of the gene of interest.[7] On the other hand,
this technique has allowed researchers to develop
methods that can help in the development of
transgenic animals and plants which is one of the
highlighted parts and can be used widely by the
researchers to get desired results. However, scientists
are working hard to understand the principles and
objectives of gene-editing technology to get results
and therapeutic results so that various diseases can be
treated and cured.
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The extravagant creation of genome altering has
changed science on the human genome in recent
years, assisting scientists with bettering clarify the
commitment of a solitary quality item to a life form's
sickness. The approach of hereditary designing (DNA
or RNA control) made another outskirt in genome
altering during the 1970s. Genome altering
advancements have been created at a quick rate in
recent years dependent on designed or bacterial
nucleases and have begun to exhibit astounding
utility in various fields, going from essential science
to applied biotechnology and biomedical exploration.
In vitro or in vivo genome altering can be refined by
giving the in situ altering apparatus that proficiently
embeds, removes, and 'rectifies' qualities just as
behaviors other exceptionally specific genomic
adjustments.
Directed DNA alterations start with the creation of
twofold abandoned nuclease-prompted breaks
(DSBs), prompting the actuation of exceptionally
proficient cell DNA recombination pathways in
mammalian cells. Both of the two significant
pathways that exist in practically all phone types and
species can reestablish nuclease-prompted DNA
DSBs: homology-coordinated fix (HDR) and
nonhomologous end-joining (NHEJ), bringing about
focused consolidation or genome disturbance.
Customarily, homologous recombination (HR), in
which flawless homologous DNA sections are
utilized as models, has been the technique for
acknowledging Targ Targeted nucleases have been
found as an elective strategy to improve the
effectiveness of HDR-interceded hereditary alteration
after it was found that DSBs could expand the
occurrence of HDR by numerous significant degrees.
This strategy and its usage are known to be easier
than HR-based methodologies because a) there is no
requirement for a maintenance grid and (b) the type
of cell has less impact on the adequacy of adjustment
(as opposed to HR, NHEJ might be dynamic all
through the cell cycle). Subsequently, like RNAi,
NHEJ can be actualized in deified cell lines to create
the inactivation of a solitary quality or a few
qualities, however by causing loss-of-work
transformations, it might prompt irreversible quality
inactivation. In the early improvement phase of
genome altering, to instigate the necessary DSBs at
every particular DNA target site, the designing of
unmistakable zinc-finger nucleases (ZFNs) or Megan
These nuclease frameworks required progressed skill
to deliver fake proteins made out of adjustable
succession explicit DNA-restricting spaces, each
connected to an objective cleavage vague nuclease,
giving analysts unmatched hereditary control
strategies.
V. SIGNIFICANCE OF GENE EDITING
Gene editing has provided scientists a platform
that can help them to make physiological changes to
see phenotypic results. Such physical change can
give rise to change in eye color. Researchers can cut
the DNA at specific sites with the help of nuclease
and can join the fragments with the help of biological
scissors which are widely used in the process of gene
editing. Various diseases can be treated by altering
the gene to get the desired result.[12] Most of the
researchers are working on the functioning of
CRISPR/Cas9 technology in treating cancer because
it allows them to stop the proliferation of the
cancerous cells and it also provides them a good
scope to treat other diseases like diabetes and cystic
fibrosis.[9] However, other than the significance, it
has various challenges that are related to the working
of the gene-editing techniques.
VI. ADVANTAGES OF GENE EDITING
With the help of biotechnology, the field of
science and technology has emerged and elevated to
establish various approaches that can help researchers
to develop new things and experiments. The
implementation of the techniques that are required to
modify the genome of the living organism has
allowed many researchers to develop different
approaches in the areas of agricultural science, in the
management of disease, in the emergence of
biomedical science, and the development of
environmental science. With the help of gene editing
tools and technologies, various fields and areas have
been covered that can help in the generation of
precautions and methods in the treatment of different
types of disease such as rating cancers, hematology,
transplantation of the organ, dermatology, neurology,
and hepatology. Many fields of science and
technology are working on the significance of such
technology to deliver new approaches in the
treatment of such chronic and life-taking
diseases.[11] The mitigation property of the gene-
editing tools and techniques has helped millions of
patients to get treated successfully. The mechanism
and experiments associate with the use of
CRISPR/Cas9 has allowed altering the
conformational changes in the T-cells to identify the
tumor cells and to destroy the abnormal cells that are
associated with the proliferation of cancer cells.
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Through this process, various researchers and
scientists got the opportunity to treat and kill cancer
cells to save millions of patients that suffers from the
deadly disease. With the help of such innovative
technology and tools, other deadly diseases such as
blood disorder, Parkinson's disease, AIDS, and
diabetes can be treated, and various success reports
have been reported with the use of CRISPR/Cas9
technology. This process can allow various
researchers to develop new drugs that can help them
to cure another genetic disease that has no other
pathway although, the process of CRISP/Cas9 has
allowed many researchers to develop robust
strategies and planning to generate more medicines
and other innovative tools.
VII. CHALLENGES IN GENE EDITING
TECHNIQUE
One of the major challenges that are related to
gene-editing technology is related to the technical
issues that arise during the engineering of the DNA
of interest. Various ethical concerns are related to the
implementation of gene-editing techniques as well as
bioethical issues that can affect the working of the
gene-editing technology. Moreover, the process is
complex, and it includes various ethical as well as
technical issues that can affect the working of the
gene-editing technology. However, the main
disadvantage associated with the use of gene editing
technologies includes ethical issues that have affected
the emergence of gene-editing technology. Moreover,
the implementation of such tools and technologies
has resulted in the development of mutation that can
provide other side effects when such tools and
techniques are administered without proper protocol.
Also, the major disadvantage of such tools and
technology is that the process is very expensive as
well as the reagents that are used in the process is not
easily available as well as very costly.
VIII. CONCLUSION
From this report, it can be concluded that the
technologies of gene editing provide a great platform
for researchers to develop a therapeutic agent to treat
the disease that is mainly generated through genetic
disorders. With the help of such innovative
technologies, researchers can develop new genomes
that can be used in the treatment of different kinds of
diseases especially cancer. The process is executed
by cutting the DNA sequence with specific enzymes
and thus this phenomenon is referred to as nuclei
engineering and through this process, the DNA can
be modified and can be further inserted into the gene
of interest.
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