GENOME EDITING

- Genome editing (also called gene editing) is a group of technologies that give scientists the ability to change an
organism's DNA. These technologies allow genetic material to be added, removed, or altered at particular
locations in the genome.
- Genome editing is of great interest in the prevention and treatment of human diseases.
- Ethical concerns arise when genome editing, using technologies such as CRISPR-Cas9, is used to alter human
genomes. Germline cell and embryo genome editing bring up a number of ethical challenges, including whether it
would be permissible to use this technology to enhance normal human traits (such as height or intelligence).
Based on concerns about ethics and safety, germline cell and embryo genome editing are currently illegal in many
countries.

2. CRISPR/CAS-9
• What is it?
- CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) technology has emerged as a powerful
technology for genome editing and is now widely used in basic biomedical research to explore gene function.
- Researchers, built upon recent biochemical studies describing a bacterial acquired immunity-like system that
relies on Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) to cleave. CRISPR units form
the backbone of this protection system.

• Where it is used?
- The potent ability of CRISPR/Cas9 system to inhibit the expression of targeted gene is being exploited as a
new class of therapeutics for a variety of diseases.
- CRISPR/Cas9 (CRISPR-associated protein 9) genome editing has been used to correct disease-causing DNA
mutations ranging from a single base pair to large deletions in model systems ranging from cells in vitro to
animals in vivo.
- Bacteria use CRISPR-associated (Cas) proteins to cleave foreign DNA into short fragments that are
subsequently incorporated into CRISPR. These DNA fragments are then transcribed into RNA and are
incorporated into a complex with other Cas proteins, including the Cas9 endonuclease. When foreign DNA
homologous to a particular CRISPR RNA– Cas9 complex enters the bacterial cell, the complex recognizes it
and triggers its cleavage.

• Recent advancement
ADVANTAGES
- “Practically, it's not only easier to design an oligonucleotide, it's a lot cheaper to synthesize. “ said Feng
Zhang, lead investigator on one of the Science papers and a member of the Broad Institute. The simplicity
offered by the new approach could fuel uptake and acceptance of genome-editing approaches throughout the
scientific community.
- The simplicity of the type II CRISPR nuclease, with only three required components (Cas9 along with the
crRNA and trRNA) makes this system amenable to adaptation for genome editing. This potential was realized
in 2012 by the Doudna and Charpentier labs
- The CRISPR/Cas system is by far the most user friendly. It is now also clear that Cas9’s potential reaches
beyond DNA cleavage, and its usefulness for genome locus-specific recruitment of proteins will likely only be
limited by our imagination.

(c) Diseases involved

- More recently, this technology has been increasingly applied to the study or treatment of human diseases,
including Barth syndrome effects on the heart, Duchenne muscular dystrophy, Chronic granulomatous disease,
hemophilia, β-Thalassemia, and cystic fibrosis.
 - In addition to genetic diseases, CRISPR/Cas9 gene editing has also been applied in immunology-focused
applications such as the targeting of C-C chemokine receptor type 5, the programmed death 1 gene, or the
creation of chimeric antigen receptors in T cells for purposes such as the treatment of the acquired immune
deficiency syndrome (AIDS) or promoting anti-tumor immunotherapy.
- Cationic liposomes were linked to aptamers by the post-insertion method and were used to deliver therapeutic
CRISPR/Cas9 that target the survival gene, polo-like kinase 1, in tumor cells. Furthermore, silencing promoted
a conspicuous regression of prostate cancer in vivo.

REFERENCES:
• Cain, C. (n.d.). CRISPR genome editing. doi:10.1038/scibx.2013.77
• What are genome editing and CRISPR-Cas9? 12. (2018). In Help Me Understand Genetics Genomic Research. Retrieved from https://ghr.nlm.nih.gov/
• Wang, H., Russa, M., & Qi, L. S. (2016). CRISPR/Cas9 in Genome Editing and Beyond. doi:10.1146/annurev-biochem-060815-014607
• Reis, A., B., Homblower, B., Robb, B., G., & B. (2014). CRISPR/Cas9 & Targeted Genome Editing: New Era in Molecular Biology. (1). Retrieved from
https://international.neb.com/tools-and-resources/feature-articles/crispr-cas9-and-targeted-genome-editing-a-new-era-in-molecular-biology
• Zhen, S., Takahashi, Y., Narita, S., Yang, Y., & Li, X. (n.d.). Targeted delivery of CRISPR/Cas9 to prostate cancer by modified gRNA using a flexible aptamer-
cationic liposome. doi:10.18632/oncotarget.14072
• Cai, L., Fisher, A. L., Huang, H., & Xie, Z. (2016). CRISPR-mediated genome editing and human diseases. Genes & diseases, 3(4), 244-251.
doi:10.1016/j.gendis.2016.07.003