Application of CRISPR Technology in the Treatment of Diabetes

Rui Cai

, Ming Cong

, Yanzhi Liu

and Chenyu Xue

College of Biological & Pharmaceutical Sciences, China Three Gorges University, Yichang, China

School of Life Sciences, Southern University of Science and Technology, Shenzhen, China

College of Life Sciences, Chongqing Normal University, China

College of Life Sciences and Technology, Beijing University of Chemical Technology, Beijing, China

Keywords: CRISPR‑Cas9, Diabetes Mellitus, Gene Therapy.

Abstract: The prevalence of diabetes is high worldwide (more than 800 million adults worldwide have diabetes). At

present, CRISPR technology therapy is the best treatment effect and the most promising method. It can

fundamentally solve the pathogenesis of diabetes by editing the WFS1 gene or knocking out the RNLS gene

in pluripotent stem cells. This paper studies the causes of diabetes and proposes that CRISPR can realize the

reversal of diabetes, which may have certain implications for the treatment of type 2 diabetes (T2D) and other

diseases in the future. Noncoding genomic defects in diabetes are analyzed and CRISPER technology is used

to reveal a diabetic regulatory circuit. Finally, the pathogenesis of T1D and T2D were analyzed, and new

treatment strategies for type 1 and T2D were revealed, which provided references for future CRISPR

application research in diabetes. However, there are still problems to be solved, such as technical research

limitations, potential safety risks, and the balance between technical application and ethics. Future research

can also focus on the combination of CRISPR and other emerging technologies (such as nanotechnology,

gene therapy, etc).

1 INTRODUCTION

Diabetes is a chronic metabolic disease characterized

by elevated blood sugar due to insufficient insulin

secretion or insulin resistance. It can be divided into

type 1 diabetes (T1D) and type 2 diabetes (T2D)

according to different pathogenesis and clinical

manifestations. Due to its serious harm to public

health, it has attracted the attention of domestic and

foreign researchers in recent years. According to

relevant reports, the global prevalence of diabetes has

increased significantly in the past 30 years. From

1990 to 2022, the number of adult diabetes patients

over 18 years old in the world will surge from about

200 million in 1990 to 828 million (Zhou, et al.,

2024).

With the development of medical technology,

many new methods for treating diabetes have

emerged, among which CRISPR is widely considered

to be the most effective and promising method.

CRISPR is a gene editing technology, which is

regarded as an efficient and accurate treatment

method for diabetes in recent years, and shows great

application potential. It can fundamentally solve the

pathogenesis of diabetes by editing WFS1 gene or

knocking out RNLS gene in multi-functional stem

cells. Traditional treatment methods generally rely on

medication to control the patient's condition, with a

focus on external regulation of blood sugar; CRISPR

can repair the internal causes of diabetes from the

genetic level (Maxwell, et al., 2020), and at the same

time achieve personalized treatment to improve the

treatment effect. It has significant advantages and

broad prospects for development. If they can achieve

coordinated development, they will be more and more

widely used in the treatment of diabetes.

This study summarized the principle and

application achievements of CRISPR in the treatment

of diabetes at home and abroad in recent years,

including editing WFS1 gene in multi-functional

stem cells, up regulating mRNA of transcription

factor HNF1A, and knocking out kidney enzyme

protein gene and other therapeutic strategies to

eradicate diabetes. The principles of these

technologies were elaborated in detail, and their

practical application level and potential negative

effects were analyzed. Corresponding strategies have

been proposed to address technical issues such as

Cai, R., Cong, M., Liu, Y. and Xue, C.

Application of CRISPR Technology in the Treatment of Diabetes.

DOI: 10.5220/0014399300004933

Paper published under CC license (CC BY-NC-ND 4.0)

In Proceedings of the 1st International Conference on Biomedical Engineering and Food Science (BEFS 2025), pages 64-68

ISBN: 978-989-758-789-4

gene editing instability and off target effects, as well

as ethical concerns. At the same time, we compared

the traditional treatment of diabetes with CRISPR,

and analyzed their advantages and disadvantages.

2 INTRODUCTION OF CRISPR

The CRISPR-Cas system provides adaptive

immunity to bacteria and archaea. In the adaptation,

Cas1-Cas2 inserts the original spacer from the foreign

genetic element into the CRISPR array as a new

spacer (represented as a rectangle of a different

color), separated by CRISPR repeats (represented as

a blue diamond). During crRNA biogenesis, CRISPR

arrays are transcribed into pre-crRNAs, which are

processed into mature crRNAs, each with an interval.

crRNA (crRNA- transactive crRNA (tracrRNA)) is

assembled with effector proteins or complexes to

form a monitoring complex that recognizes and

degrades foreign genetic elements that complement

the crRNA interval during interference. The Type I

CRISPR system is further subdivided into three seed

types. They were based on the degree of homology

between Cas9 proteins and the presence or absence of

additional Cas proteins involved in adaptation. Type

II-A and type II-B systems include Csn2 and Cas4,

respectively, while most type II-C systems are

characterized by the lack of Cas4 and Csn2II-C

CRISPR arrays that embed internal promotors in each

repeat sequence, generating pre-nested crRNA as a

source of mature crRNA. Instead of processing a

single pre-CRRNA transcript, another characteristic

of all type II Cas9s is the need for PAM (progasm

neighbor motif) sequences in the target DNA.

3 DIABETES

3.1 Introduction to Diabetes

Diabetes mellitus is a group of disorders of

carbohydrate, protein and fat metabolism caused by

absolute or relative insufficiency of insulin secretion

and/or insulin utilization disorders, with

hyperglycemia as the main sign. The causes of the

disease are diverse and can be roughly divided into

the following: ethnic and genetic factors, long-term

overfeeding, obesity factors, mental factors and

autoimmune. Its clinical manifestations can also be

divided into T1D and T2D. In general, T1D occurs

more quickly, and the disease is often in adolescents

and children. T2D has no obvious "three more and

one less" symptoms, and is often a chronic disease, it

is not easy to distinguish when and why the onset, and

sometimes patients have been ill and failed to find

treatment in time, only patients can be confirmed by

blood sugar test.

3.2 Traditional Treatment of Diabetes

In the traditional treatment of diabetes, insulin

replacement therapy is the main choice for patients

with T1D, and changes in diet and lifestyle are

considered to be the best choice for treatment and

management of T2D (Bastaki, et al., 2005). Insulin is

also important in the treatment of T2D when blood

sugar levels cannot be controlled through diet, weight

loss, exercise and oral medication.

3.2.1 Dietary Therapy and Exercise

Therapy

The main treatment approach of dietary therapy is to

accurately calculate the patient's daily calorie

requirements and then control their diet to maintain

blood sugar balance. Generally, the appropriate

calorie intake value needs to be determined based on

the patient's age, gender, weight, physical activity

level, and other factors. For example, a 50-year-old

male diabetes patient who is engaged in light physical

labor, 170 cm tall and 75 kg in weight, is estimated to

need about 1800 kcal of heat per day according to the

formula. Among them, carbohydrate accounts for

50% -65%, protein accounts for 15% -20%, and fat

accounts for 20% -30%. It focuses on unsaturated

fatty acids and reduces animal fat intake (diabetes

Branch of the Chinese Medical Association, 2021).

Aerobic exercises such as brisk walking, jogging,

and swimming can all be the first choice for exercise

therapy. During aerobic exercise, muscles continue to

contract, thereby increasing the uptake and utilization

of glucose in the blood; At the same time, exercise

enhances insulin sensitivity, allowing insulin to

function more efficiently and guiding glucose into

cells for energy supply, resulting in a decrease in

blood sugar levels. It is worth noting that patients

with complications of diabetes need to exercise

carefully to avoid additional damage to the body. For

example, patients with retinopathy should avoid

eyeground bleeding caused by intense exercise, and

patients with neuropathy should prevent foot injury.

Application of CRISPR Technology in the Treatment of Diabetes

3.2.2 Drug Therapy

In the treatment of diabetes, the drugs used are

generally divided into oral hypoglycemic drugs and

insulin injection. The former is mainly used to treat

T2D, and the latter is used to treat T1D. For T2D

patients, insulin is also needed when oral drugs

cannot control blood sugar. Oral hypoglycemic drugs

include sulfonylureas, biguanides, alpha glucosidase

inhibitors, etc.

The most common treatment drug is metformin,

which is the first-line drug for T2D. Under normal

circumstances, the liver can convert non sugar

substances into glucose and release it into the

bloodstream, while metformin can interfere with this

pathway, reducing liver glucose output and

subsequently lowering fasting blood glucose levels.

On the other hand, metformin can enhance the

sensitivity of peripheral tissues (such as muscles and

adipose tissue) to insulin. Insulin is like a "key" to

open the "door" for cells to absorb glucose. However,

in patients with diabetes, especially in patients with

T2D, the role of this "key" is often blocked. Cells are

not sensitive to insulin, and it is difficult for glucose

to enter cells for energy supply. Metformin can

improve this insulin resistance state, allowing insulin

to function better, promoting muscle cells to uptake

glucose from the blood for energy metabolism, and

also increasing the utilization of glucose by adipose

tissue, thereby effectively reducing blood sugar. Both

postprandial blood sugar and overall daily blood

sugar levels can be well regulated.

For patients with diabetes, when their islet function

is impaired, they cannot secrete enough insulin, or the

body is resistant to insulin, and they cannot

effectively use insulin, they need to inject exogenous

insulin to make up for this deficiency. Insulin can

bind to specific insulin receptors on the surface of

muscle cells and adipocytes, activating a series of

signaling pathways within the cells, allowing glucose

from the blood to enter the cells smoothly, providing

energy to the cells and reducing the glucose content

in the blood. Insulin not only inhibits glycogen

breakdown and gluconeogenesis in the liver, but also

stimulates liver and muscle cells to synthesize excess

glucose in the blood and store it as glycogen,

ultimately achieving the goal of lowering blood

sugar.

3.3 Relationship between CRISPR and

Treatment of Diabetes

CRISPR, as an accurate gene editing tool, has made

great contributions to the exploration of the

pathogenesis of diabetes. Researchers can use it to

knock out and modify specific genes in cells, so as to

analyze the role of genes related to insulin resistance

and abnormal pancreatic β cell function in the

pathogenesis of diabetes.

For T1D, the patient's own islet beta cells are

damaged due to the attack of the immune system. The

researchers imagine to use CRISPR to edit the

patient's immune cells so that they will not attack the

islet beta cells by mistake, or to convert induced

pluripotent stem cells (iPSCs) into functional islet

beta cells through gene editing, and then transplant

them back into the patient's body (Staedtke, et al.,

2020) to restore normal insulin secretion. For T2D,

this technology can be used to correct the genetic

defects that lead to insulin resistance, enhance the

sensitivity of the body to insulin (Venkatrama, et al.,

2024), and improve the disease from the root.

According to existing research, the traditional

treatment of diabetes focuses on regulating blood

sugar from the outside, such as drugs to stimulate

insulin secretion, supplement insulin or improve

insulin sensitivity, while CRISPR is committed to

repairing the internal root cause of diabetes from the

genetic level (Bora, et al., 2023). If the two can

develop together, they may provide more accurate

and thorough treatment paths for diabetes patients in

the future.

4 APPLICATION OF CRISPR IN

THE TREATMENT OF

DIABETES

4.1 Realization of Reversal of Diabetes

Diabetes may be caused by gene mutation caused by

acquired factors. It is possible to reverse diabetes

through CRISPR. A mutation in the WFS1 gene in

patients with Wolfram syndrome 1 causes a rare

inherited form of insulin-dependent diabetes. To

develop a treatment for this form of diabetes, Dr.

Jeffrey R. Millman, assistant professor of medicine

and biomedical engineering at the University of

Washington, and his team selected cells from a patient

with Wolfram syndrome 1 (WFS1) and derived them

to induce and differentiate into pluripotent stem cells

BEFS 2025 - International Conference on Biomedical Engineering and Food Science

(IPscs). CRIPSR/Cas9 was then used to edit the WFS1

gene in patient-derived IPscs to produce autogene-

corrected SC-β. The researchers then compared the

gene-edited cells with the same batch of insulin-

secreting beta cells that had not been CRISPR-edited.

In mice with severe diabetes, the CRISPR-edited,

newly grown beta cells secreted glucose more

efficiently. The CRISPR-edited cells placed under the

skin made diabetes disappear quickly in mice, and the

animals' insulin secretion improved and their blood

sugar stayed within normal ranges during a six-month

monitoring period. CRISPR was first used to repair a

genetic defect that caused diabetes in patients and

successfully reversed diabetes (Maxwell, et al., 2020).

The research extends a strategy for tackling T1D

and may have implications for the treatment of T2D

and other diseases in the future.

Looking forward to the future, the researchers

extract cells from human skin for differentiation, if the

use of human metabolite cells for experiments, not

only reduce the difficulty of raw material acquisition

and reduce the cost of the experiment, in addition, the

success of the experiment may also help solve other

complications of Wolfram syndrome 1 (WFS1)

patients.

4.2 Reveal a Regulation Circuit of

Diabetes

The onset of diabetes is often caused by the adverse

effects of multiple factors, rather than the loss of a

single isolated factor. The study found that HNF1A

homeobox A (HNF1A) encodes the homeodomain

transcription factor, whose haploid under-dose

mutation induces diabetes. HNF1A's antisense

lncRNA promoter, HASTER, can cis-regulate

HNF1A transcription and maintain the cell-specific

physiological concentration of HNF1A through

positive and negative feedback loops. HASTER

mutant mouse pancreatic beta cells exhibit HNF1A

silencing or overexpression, which leads to

hyperglycemia. After upregulation of Hnf1a mRNA

by about 30-80% through the use of CRISPR-Cas9,

Haster RNA increased by about 50-120%, revealing

the mechanism of action of HASTER promoter as a

regulator of HNF1A, with a negative feedback loop

between them. That is, HNF1A positively regulates

HASTER, while HASTER negatively regulates

HNF1A, a finding that contributes to the development

of novel diabetes treatments and facilitates our

understanding of noncoding genomic defects in the

disease (Beucher, et al., 2020).

In the future, there may be synthetic HASTER

isofactor introduced into human body for treatment,

which can not only be used as a regulatory factor of

HNF1A, but also as a cofactor of other regulatory

factors. It also provides clues to the genetic

mechanism of diabetes.

4.3 Reveal New Treatment Strategies

for T1D and T2D

T1D is mainly caused by immune system

abnormalities that lead to the destruction of islet beta

cells. CRISPR has been used to study genes

associated with beta cell function. For example,

CRISPR genome-wide screening revealed the effect

of the RNLS (Renalase) gene on beta cell survival.

Studies have found that knocking out RNLS can

significantly reduce the sensitivity of beta cells to

oxidative stress, thereby reducing the risk of T1D

development (Cai, et al, 2020). At the same time, the

research team further identified an already FDA-

approved drug, Pargyline, whose mechanism of

action is to bind to RNLS and inhibit it. The study

also showed that Pargyline has good safety while

protecting β cells and preventing diabetes in mice,

and has considerable development significance in the

future.

In addition, CRISPR has also been used to modify

immune cells to suppress autoimmune responses

against beta cells, providing new ideas for

immunotherapy of T1D (Bevacqua, et al. 2021).

The team also established a genetic model in

human islet cells and revealed the regulatory and

genetic mechanisms by which a non-coding variation

is associated with diabetes risk in humans.

The pathogenesis of T2D is complex, involving

insulin resistance and beta cell failure. CRISPR is

widely used to screen for genetic risk genes

associated with T2D. For example, using CRISPR

screening, it was found that the CALCOCO2 gene

plays a key role in the regulation of beta cell function,

and gene mutations are closely associated with T2D

risk, and the experiment also completed a genome-

wide CRISPR hybrid screening, providing a

comprehensive perturbation dataset for future

CRISPR research (Rottner, et al., 2023).

In addition, CRISPR has been used to edit fat cells

to improve metabolic function and thus reduce the

risk of T2D by promoting Browning of white fat

(Tsagkaraki, et al., 2021), and also provides a future

cellular therapy strategy for metabolic diseases,

especially in larger animals.

Application of CRISPR Technology in the Treatment of Diabetes

5 CONCLUSION

CRISPR has demonstrated significant potential in the

field of diabetes treatment, offering more options for

patients. By precisely repairing genetic defects and

inducing cell transformation, CRISPR holds promise

for treating diabetes at its root cause and enabling

personalized therapeutic approaches. Currently,

CRISPR has achieved remarkable results in animal

experiments, and some clinical trials are steadily

progressing too. These make a credible foundation for

its clinical application.

However, challenges and limitations remain in the

application of CRISPR for diabetes treatment.

Technically, issues such as off-target effects and

instability in gene editing during the treatment

process may impact the efficacy of diabetes therapy

and even interfere with normal cellular physiological

functions. In terms of safety, the Cas protein in the

CRISPR system which derived from bacteria may be

recognized as a foreign entity by the human immune

system and trigger immune responses when

introduced into human cells for gene editing.

Ethically, gene editing technology continues to face

significant controversy. Additionally, CRISPR-based

diabetes treatments are still in the research and

development phase, with high associated costs. This

may result in only a small number of economically

capable patients being able to afford such treatments

in the future and this may exacerbate inequalities in

the distribution of medical resources.

It can be seen that in the near future, collaboration

among experts in biology, medicine, ethics, and law

will be essential to advance CRISPR in diabetes

treatment. It is believed that with continuous

technological improvements and breakthroughs,

CRISPR will bring revolutionary changes to diabetes

therapy, offering genuine hope to hundreds of

millions of diabetic patients worldwide and enable

them to overcome the challenges of diabetes to regain

a healthy life.

AUTHORS CONTRIBUTION

All the authors contributed equally and their names

were listed in alphabetical order.

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