Jellyfish GFP Uses for Gene Editing
The jellyfish Aequorea victoria produces a Green Fluorescent Protein (GFP, for short), which glows when exposed to light in the blue to ultraviolet range. Purified GFP solutions appear yellow under typical indoor lighting, but they glow with a bright green hue in the sunlight. The protein absorbs ultraviolet radiation and then emits it as lower-energy green light. Its usage in the world of genetic engineering and molecular cloning has garnered the attention of scientists and laymen alike, and its versatility is noteworthy. It is a biological marker that can monitor physiological processes, visualize protein localization, and detect transgenic expression in vivo (in live, isolated cells).
The protein is made up of 238 amino acids and three of them (Numbers 65 to 67) form a structure that emits visible green fluorescent light. The protein can attach to and mark other proteins with fluorescence, which enables scientists to mark the presence of another particular protein in organic structures. Gfp refers to the gene that produces the green fluorescent protein (GFP). Using DNA recombinant technology (usually the molecular cloning of foreign DNA into bacterial extrachromosomal DNA elements), scientists combine the Gfp gene to another gene that produces a protein that they want to study, and then they insert the complex into a cell (usually an Escherichia coli cell). If the cell fluoresces under blue light, scientists may infer that the cell expresses the other gene too. In essence, GFP can label specific organelles, cells, and tissues. Since the GFP gene is heritable, the descendants of labeled entities express it. This is helpful in DNA repair bioassays.
An example where GFP can be useful is in studying the effectiveness of RAD52, a gene that has a role in repairing damaged DNA. Since deficiencies in DNA repair mechanisms appear to be an underlying cause of cancer, RAD52's potential for use in cancer treatments is being heavily researched. But what’s the method for marking a gene in molecular cloning?
Firstly, restriction enzymes that can isolate the gene of interest need to be chosen. Restriction enzymes include EcoRI, EcoRII, BamHI, HindIII, etc. When they come into contact with a DNA sequence with a shape that matches a part of the enzyme, called the recognition site, they induce a break in both strands of the DNA molecule. Each restriction enzyme recognises a specific recognition site or DNA sequence.