Within the advancement of modern technology, a great revolution is taking place in genetic engineering as well. The applications of this technology are limitless. CRISPR-Cas9 is a unique technology that enables geneticists and medical researchers to edit parts of the genome by removing, adding or altering sections of the DNA sequence. It is currently the simplest, most versatile and precise method of genetic manipulation and is therefore causing a buzz in the science world.
What is CRISPR
CRISPR technology is a simple but powerful tool for editing genomes. It allows researchers to easily modify DNA sequences and alter gene activation. Many of its potential applications include correcting genetic defects, preventing the spread of disease, and improving crops. In popular usage, "CRISPR" is an acronym for "CRISPR-Cas9". CRISPRs are specific extensions of DNA. CRISPR stands for "Short Polyndrochromic Repetitions with Regular Clusters". CRISPR genome engineering technology allows scientists to easily and accurately edit the DNA of any gene. Protein Cas9 is an enzyme that acts like a pair of molecular scissors capable of cutting the strands of DNA. CRISPR technology was adapted from the natural defense mechanisms of bacteria and archaeologists. These organisms use a variety of CAS proteins, including CRISPR-derived RNA and CAS 9, to suppress attacks by viruses and other foreign bodies. They do so primarily by cutting and destroying the DNA of a foreign invader. These compounds allow genes to be manipulated or "edited" when transferred to other complex organisms. In nature, CRISPR polyandromic replication plays an important role in microbial immunity. When a virus infects a microbial cell, the microbe uses a special CRSIPR-linked nucleus (CAS9) to cut a piece of viral DNA. The nucleus is directed to its target sequence by a short piece of RNA called the guiding RNA, which complements the target segment of the virus genome.Components of CRISPR/Cas-9
Components of CRISPR/Cas-9 based on the structure and activity of CAS-proteins, the CRISPR / CAS system can be divided into two classes. First-class systems consist of multiple subunits of cas-protein complexes, while second-class systems use single-Cas-proteins. The structure of the CRISPR / Cas-9 type is relatively simple and has been well studied and widely used in genetic engineering. Guided RNA (GRNA) and CRISPR-associated (CAS-9) proteins and CAS-9 system are two essential components of CRISPR. The CAS-9 protein, the first CAS protein used in gene modification, was derived from the Streptococcus pyogen (SPCS-9). It is a large (amino acid 1368) multi-domain DNA endonuclease that is responsible for inducing targeted DNA dual fiber breakage and is known as genetic shears.Mechanism of CRISPR/Cas-9 genome editing
The CRISPR / Cas-9 gene editing mechanism can generally be divided into three stages: acceptance, breakdown, and repair. The designed SGRNA directs CAS-9 and identifies the target sequence of the gene of interest through its complementary base pair component. . The CAS-9 protein remains inactive in the absence of SGRNA. The CAS-9 nucleus undergoes bilateral fractures at two locations. The PAM sequence is below the DNA sequence at the incision site and varies in size depending on the bacterial species. Identifies the sequence of nuclear CAS-9 proteins commonly used in the genome editing tool. Once CAS-9 finds a target site with a suitable PAM, it triggers local DNA fusion and then RNA-DNA hybridization, but the sequence of targeted DNA on how CAS-9 enzyme dissolves is still unclear. The CAS-9 protein is then activated to break down DNA. The complementary DNA is then produced.Applications of CRISPR technology
Cell and gene therapy- CRISPR is poised to revolutionize medicine and has the potential to cure a number of genetic diseases, including neurodegenerative diseases, blood disorders, cancer and eye disorders. As an example, CRISPR can also be used to generate chimeric immunosuppressive antibody (CAR) T cells, an immunotherapeutic agent used to treat cancer. Chimikic antibodies are designed to release antibodies before the T cells are extracted from the patient and injected back into the body. Receptors allow T cells to target more efficiently and destroy specific cancers that affect the patient. Although we are still in the early stages of clinical trials, this technology could be used to treat thousands of genetic conditions in the future. Diagnostics- During the COVID 19 pandemic, CRISPR was used as a potential therapeutic tool as well as a corona virus detection tool. Mammoth Biology has also developed a CRISPR-based COVID-19 diagnostic method called DETECTR. Like Sherlock and Stopcovid, Detector also uses the CAS9 search function to use naturally occurring CAS nuclei, such as CAS12 and CAS13, to identify genetic material from viruses. Similar diagnoses have been made using the search function in case 9 to identify other infectious and genetic diseases. Agriculture -Genetically modified technology has great potential in agriculture, and experts suggest that CRISPR modified foods may be available in 5-10 years. This is mainly due to the fact that it can be used to grow disease resistant and drought tolerant crops. It can also be used to increase the shelf life of other digestible foods, reduce food contamination and provide healthier food at a relatively low cost.Bioenergy - The focus on biofuels as a viable alternative to fossil fuels has been around for some time now. However, there are several barriers to scale biofuel production. Using CRISPR, scientists have recently been able to make significant progress in this area. For example, the multiplication factors that control lipid production in algae have led to a large increase in lipid production for biodiesel production. Also, genetic modification can increase the ability of yeasts to withstand harsh conditions during biofuel production. It has also increased the editing efficiency of the bacterial species used to produce ethanol. There is no doubt that CRISPR-Cas9 has revolutionized in genetic engineering. However, we are beginning to see the benefits and capabilities of this incredible technology. The number of biotechnology start-ups focusing on CRISPR-Cas9 gene editing technology is also growing, and many researchers are looking for new ways to apply this technology to solve real-world problems, including epigenome editing, new cells and gene therapies, and infectious diseases
