What is CRISPR?
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a revolutionary genetic engineering technology that allows scientists to make precise changes to the DNA of living organisms.
What can CRISPR do for scientists?
There are a wide range of applications CRISPR is used within to benefit scientists and experts, listed below are some of the most common fields where CRISPR is used to benefit researchers:
Gene therapy: CRISPR can be used for gene therapy by editing the DNA sequence of a patient's cells to correct or introduce a desired genetic trait. This is done by using the CRISPR system to target and cut the specific DNA sequence, allowing for the insertion or deletion of genetic material. The edited cells can then be reintroduced into the patient's body to treat a genetic disorder.
Drug discovery: CRISPR can be used for drug discovery by creating disease models using cells with edited genes to study the effects of potential drugs on specific genetic mutations. This allows for a more targeted approach to drug discovery and personalised medicine.
Agricultural biotechnology: CRISPR can be used in agricultural biotech to produce crops with desirable traits such as increased yield, improved disease resistance, and enhanced nutrition. This is done by using the CRISPR system to edit the genes responsible for these traits in plant cells.
Basic research: CRISPR is used extensively in basic research to study the function of genes and their role in various biological processes. Researchers can use CRISPR to create knockout or knock-in models of genes to observe their effects on cells and organisms. CRISPR can also be used to develop animal models of human diseases to study disease progression and test potential therapies.
CRISPR Techniques and Protocols
Cas9-based CRISPR: This is the original and most well-known version of CRISPR, which uses the Cas9 protein to cut DNA at the target site specified by the gRNA. This version of CRISPR is used for a wide range of applications, from gene knockout to gene insertion.
Take a look at relevant publications regarding this protocol below:
Genome Engineering with Cas9 - Doudna, J.A. and Charpentier, E., 2014. The new frontier of genome engineering with CRISPR-Cas9. Science, 346(6213),
Cas9 and its work with cancer - Wang, S.W., Gao, C., Zheng, Y.M., Yi, L., Lu, J.C., Huang, X.Y., Cai, J.B., Zhang, P.F., Cui, Y.H. and Ke, A.W., 2022. Current applications and future perspective of CRISPR/Cas9 gene editing in cancer. Molecular Cancer, 21(1), pp.1-27.
Cas12a-based CRISPR: This is a newer version of CRISPR that uses the Cas12a protein (also known as Cpf1) to cut DNA at the target site. Cas12a-based CRISPR is similar to Cas9-based CRISPR, but has some advantages such as smaller size and different target recognition properties.
Take a look at relevant publications regarding this protocol below:
Current Applications of Cas12a - Dronina, J., Samukaite-Bubniene, U. and Ramanavicius, A., 2022. Towards application of CRISPR-Cas12a in the design of modern viral DNA detection tools. Journal of Nanobiotechnology, 20(1), pp.1-15.
CRISPRi: This version of CRISPR does not cut the DNA, but instead uses a modified Cas protein to interfere with gene expression by blocking the transcription process. This approach is often used for gene knockdown experiments or to study gene function.
CRISPRa: This version of CRISPR also uses a modified Cas protein, but instead of blocking transcription it enhances it, allowing for targeted upregulation of gene expression.
Take a look at relevant publications regarding both CRISPRi and CRISPRa below:
Therapeutic Applications of CRISPRa and CRISPRi - Bendixen, L., Jensen, T.I. and Bak, R.O., 2023. CRISPR/Cas-mediated transcriptional modulation: The therapeutic promises of CRISPRa and CRISPRi. Molecular Therapy.
CRISPRi and CRISPRa for precision biology - Kampmann, M., 2018. CRISPRi and CRISPRa screens in mammalian cells for precision biology and medicine. ACS chemical biology, 13(2), pp.406-416.
How does it work?
Cas9-Based CRISPR works by using a special protein called Cas9 that can be programmed to target a specific DNA sequence. The Cas9 protein is guided to the target DNA sequence by a small RNA molecule that is designed to complement the DNA sequence of interest. Once the Cas9 protein is bound to the target DNA sequence, it can cut the DNA at that specific location. This allows scientists to either remove, replace, or add new genetic material at the targeted location.
What are some more contemporary and advanced methods of CRISPR?
For a deeper exploration of advanced topics in CRISPR, such as developing new treatments for genetic diseases by correcting or replacing defective genes, "knocking out" or "knocking in" specific genes in model organisms and engineering new therapies for cancer by targeting and destroying cancer cells; It's best to explore our dedicated webpage that goes into more depth.
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