CRISPR-associated protein 9 (Ca9) is an enzyme that uses CRISPR sequences as a guide to cleave and recognize particular strands of DNA that make the complementary sequence.
A combination of Cas9 enzymes and CRISPR sequences forms the basis of a technology known as CRISPR Ca9 that can be used to edit genes within organisms.
The problem with this technology is that when used to snip and replace DNA, a gene named p53 jumps and causes its edited cells to self-decipher- This means that when a CRISPR – Ca9 edit sticks, it may mean that p53 is not functioning correctly. A dysfunctional p53 may cause a host of cancers in the ovaries, colon, rectum, and esophagus.
In CRISPR – Ca9, unwanted genetic changes use a method that allows them to turn off gene-editing until they reach key cell cycle phases where more accurate repairs happen. A team of researchers has developed a fix to CRISPR-Cas9's that may turn out to be promising and cause ripples in Crisper Associated Cas Market.
CRISPR-Cas9 has brought in a new edge in gene editing by introducing a simpler and less expensive tool. The tool acts like scissors and can snip genetic material that one wants to alter. However, it has limited use in therapeutics as this process can also create off-target effects.
A recent study has successfully shown a more precise gene-editing, which can be considered an advancement in the Crisper Genome Editing Industry. It suppresses unintended genetic insertions, deletions, or mutations called off-target effects.
However, several methods have been developed that reported fewer problems associated with CRISPR technology. The researchers stated that such methods, though able to counter the problem, but at the same time exhibited lower editing efficiency.
The newest advanced method is used to eliminate off-target effects by using an anti-CRISPR protein known as "AcrIIA4". It is used like an "off switch" that stops SpyCas9's genome editing activity. The researchers merged AcrIIA4 with the region of human Cdt1 known as N terminal. This means that this technology will have further effects on the Crisper Industry.
Human Cdt1 is a gene that ensures replication of DNA happens only once per cell division. It works to deactivate gene editing until S and G2, phases of the cell cycle when homology-directed repair (HDR) is dominant.
There are two DNA repair processes used by organisms, HDR and Non-Homologous End Joining (NHEJ). Between the two, HDR is the preferred method because the repair can only be done with two chromosome copies in each cell. HDR uses duplicate chromosome as a template for repair. This makes gene editing furthermore precise as opposed to NHEJ. It is because NHEJ only connects the broken ends of the DNA. HDR takes place during the S and G2 stages of the cell cycle. In comparison to this, NHEJ operates in all stages, especially in G1.
This technology is a step towards resolving the problem caused by CRISPR – Ca9. However, there is a need for further development, which would ensure its usage in therapeutics.
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