جستجوی مقالات مرتبط با کلیدواژه « کریسپر » در نشریات گروه « پزشکی »

Gene editing as a molecular knife has provided a powerful tool to edit the genome inside living organisms’ bodies. Despite all the benefits, this technology has many biosecurity concerns that have prompted the international community to develop specific implementation guidelines. This study aimed to investigate the risks of gene editing technology from the point of view of biosecurity and to determine its defense strategies.

This review was conducted in the fall and winter of 2022-23. The study’s method was based on the observation and interpretation of the data obtained from relevant scientific articles and books. Scientific books and articles were searched by searching the keywords Germline Gene Editing, CRISPR, Biosecurity, CRISPR War, Biohacking, CRISPR babies, Do It Yourself, Islamic Bioethics in PubMed, Scopus, Researchgate databases and also the Google search engine was searched and checked in English .

The advent of genome editing technology has created a new paradigm in which the human genome sequence can be precisely manipulated to achieve a therapeutic effect. However, embryo editing and the design of programmed humans (super-humans) are considered one of the challenges and risks of gene editing biosecurity. Also, this technology is mentioned as a dangerous tool for biohacking and bioterrorism in the design of personalized bioweapons and emerging agents.

CRISPR-based bioweapons have destroyed the logical and strategic balance of power that has kept the world immune from using weapons of mass destruction. The world is facing a potentially more dangerous technology than nuclear weapons. As a result, establishing appropriate international and ethical laws is necessary to prevent the potential dangers of this technology and to deal with it.

Long non-coding RNAs play an important role in regulating gene expression, RNA processing, histone modification, and rearrangement of chromatin genes. These molecules can also be involved in many biological processes, such as organogenesis, cell differentiation, development, genome imprinting, quantitative compensation, and tumorigenesis. High expression of MALAT1 (a type of lncRNA) in many cancers, including breast cancer, indicates that a disorder of MALAT1 regulation is an important factor in the development of many types of cancer. Breast cancer is the most common cancer among women worldwide, and the invasion, as well as metastasis of this disease, are considered among the main causes of death. The present study aimed to knock out the MALAT1 gene in the MDA-MB-361 breast cancer cell line and evaluate its function and effects on the expression of genes associated with apoptosis.

In this study, two types of sgRNA were designed by CHOPCHOP software for exon 1 of the MALAT1 gene. These sgRNAs were cloned separately into two CRISPR vectors to generate the recombinant vectors PX459-sgRNA1 and PX459-sgRNA2. Co-transfection of these two recombinant vectors into the MDA-MB-361 cancer cell line was performed using lipofectamine 2000. MALAT1 gene editing was investigated in the cells receiving recombinant vectors. The expression of genes related to apoptosis was analyzed by Real-Time PCR. Cell proliferation and apoptosis were assessed by MTT and flow cytometry methods, respectively.

The MALAT1 gene was edited by the CRISPR method in MDA-MB-361 cells. The rate of cell proliferation in the cells of the treatment group, compared to the control groups, showed a significant decrease (P<0.05). Apoptosis levels were significantly increased in cancer cells the MALAT1 gene of which had been deleted. Moreover, the expression of BCL2 and survivin anti-apoptotic genes in treated (edited) cells was significantly reduced, compared to control cells (P<0.05). Increased expression of proapoptotic genes P53, BAK, BAX, and FAS was also observed in the edited cells (P<0.05).

The results of this study confirm that the deletion of the MALAT1 gene has a significant effect on increasing apoptosis and reducing cell proliferation. A reduction in the expression of the MALAT1 gene can prevent the growth and proliferation of breast cancer cell lines. Therefore, it seems that the control of MALAT1 oncogene expression is useful and effective for controlling tumors.

 Lung cancer is the second most common cancer in the world. The COR-L105 cell line is used as one of the most popular lung cancer cell lines in various studies. Long non-coding RNAs (lncRNAs) are a group of important cellular factors that play a key role in many processes such as growth, differentiation, replication, transcription, translation. LncRNA LINC00511 is a transcriptional regulator and has been reported to increase its expression in various cancers. The aim of this study was to knockout the LINC00511 gene using a CRISPR/Cas9 in COR-L105 cells and to investigate its effects on cancer progression and apoptosis.

 In this experimental study, two sgRNAs were designed for LINC00511 gene and cloned into two CRISPR vectors, separately. Two sgRNAs vectors were transferred to COR-L105 cells using lipofectamine. After confirmation of LINC00511 gene knockout from target cells, changes in cell proliferation and apoptosis were assayed by MTT and flow cytometry. Furthermore, the expression of genes associated with apoptosis and tumorigenesis was examined by real-time PCR.

LINC00511 gene knockout from COR-L105 cell line was performed. The expression of BCL2, survivin, EZH2, c-MYC, and MAX genes showed a meaningful decrease related to the expression in treated cells (edited) compared to the control cells (p˂0.05). Increased expression of p21 and p57 genes was also seen in the edited cells (p˂0.05). Decreased cell proliferation and increased apoptosis were observed in manipulated cells.

Knocking of LINC00511 gene in lung cancer cell line caused apoptosis and decreased cell proliferation. Therefore, inhibiting the expression of the LINC00511 gene in cancer cells leads to the control their proliferation.

Pancreatic cancer is ranked as the fourth leading cause of cancer related death. The survival rate of patients is very low and the mortality rate in men is higher than that of women. Various hereditary and non- hereditary factors are involved in the formation of pancreatic cancer; one of the important non- hereditary factors is smoking. Besides, it is known that the risk of pancreatic cancer in patients with diabetes mellitus is twice as high as in the non-diabetic population. Hereditary conditions also include genetic (activation of oncogenes and inactivation of tumor suppressor genes) and epigenetic changes (lncRNA). The mutation in KRAS oncogene is observed in 95% of the patients with pancreatic cancer. Despite the common use of ultrasound and tumor markers in diagnosing pancreatic cancer, their importance as powerful diagnostic tools has been challenged. An innovation in common therapeutic strategies such as chemotherapy, immunotherapy, and gene therapy based on genetic vectors has provided a promising tool for treatment.  Using oncolytic viruses with selective proliferation in cancer cells and minimal adverse effects has opened a new perspective on the future of treatment of this cancer. In addition, novel technologies in gene editing including CRISPR have allowed the genome manipulation of cancer cells to identify and regulate the molecular pathways involved in pancreatic cancer. Considering that multiple genetic and epigenetic changes play a role in the pathogenesis of this cancer, simultaneous control of these pathways with multiple therapies can be a useful therapeutic strategy for pancreatic cancer.