Crisper Cas9

CRISPR ( clustered regularly interspaced short palindromic repeats) is a

family of DNA sequences found in the genomes of prokaryotic organisms such
as bacteria and archea. They are derived from DNA fragments
of bacteriophages that had previously infected the prokaryote.

Site: CRISPR are found in approximately 50% of sequenced bacterial

genomes and nearly 90% of sequenced archaea.

Significance: They are used to detect and destroy DNA from similar
bacteriophages during subsequent infections. Hence these sequences play a key
role in the antiviral (i.e. anti-phage) defense system of prokaryotes and provide a
form of acquired immunity.

Cas9 (or "CRISPR-associated protein 9") is an enzyme that uses CRISPR

sequences as a guide to recognize and cleave specific strands of DNA that are
complementary to the CRISPR sequence.

Cas9 enzymes together with CRISPR sequences form the basis of a technology
known as CRISPR-Cas9 that can be used to edit genes within organisms This
editing process has a wide variety of applications including basic biological
research, development of biotechnological products, and treatment of diseases.

CRISPR/Cas9 edits genes by precisely cutting DNA and then letting natural DNA
repair processes to take over. The system consists of two parts: the Cas9 enzyme
and a guide RNA.
How does the system work?

A: CRISPR “spacer” sequences are transcribed into short RNA sequences

(“CRISPR RNAs” or “crRNAs”) capable of guiding the system to matching
sequences of DNA. When the target DNA is found, Cas9 – one of the enzymes
produced by the CRISPR system – binds to the DNA and cuts it, shutting the
targeted gene off. Using modified versions of Cas9, researchers can activate gene
expression instead of cutting the DNA. These techniques allow researchers to
study the gene’s function.
Three main categories of genetic edits can be performed with CRISPR/Cas9:

DISRUPT
If a single cut is made, a process called non-homologous end joining can result in
the addition or deletion of base pairs, disrupting the original DNA sequence and
causing gene inactivation

DELETE
A larger fragment of DNA can be deleted by using two guide RNAs that target
separate sites. After cleavage at each site, non-homologous end joining unites the
separate ends, deleting the intervening sequence
 CORRECT OR INSERT
Adding a DNA template alongside the CRISPR/Cas9 machinery allows the cell to
correct a gene, or even insert a new gene, using a process called homology
directed repair

Significance:
1. the technology can effectively correct genetic defects. Examples of such
diseases include cystic fibrosis, cataracts and Fanconi anemia. These studies
have paved the way for therapeutic applications in humans. i.e. New tools to
stop genetic diseases: Scientists are now using CRISPR/Cas9 to edit the
human genome and try to knock out genetic diseases like hypertrophic
cardiomyopathy or cystic fibrosis
2. CRISPR has been tested in early-stage clinical trials as cancer therapy, like
trying it on the BRCA-1 and 2 mutations linked to breast and ovarian
cancers.
3. As a treatment for an inherited disorder that causes blindness.
4. It's also been investigated as a strategy for preventing the spread of Lyme
disease and malaria from viral vectors to people, and it's also been studied in
animal models of HIV as a way to rid infected cells of the virus. Scientists
have even shown that CRISPR can knock HIV infections out of T cells.
 5. Powerful new antibiotics and antivirals: because we are running low on
effective antibiotics as bacteria evolve resistance to them, CRISPR/Cas9
systems could, in theory, be developed to eradicate certain bacteria more
precisely than ever (though, again, figuring out delivery mechanisms will be
a challenge).
6. One other potential application is to create gene drives, a genetic
engineering technique that increases the chances of a particular trait passing
on from parent to offspring.
7. Creating “designer babies”: This is the one that gets the most attention, and
rightly so. The concern is that there are not enough safeguards yet to prevent
harm nor enough knowledge to do definite good.

8. During the COVID-19 pandemic, the CRISPR-Cas9 system has been used to
develop various diagnostic tests for the viral infection.
Side effects: The Cas9 enzymes can occasionally “misfire” and edit DNA in
unexpected places, which in human cells might lead to cancer or even create new
diseases.