You wish to perform gene therapy to correct a mutated gene to wildtype in an adult...

You wish to perform gene therapy to correct a mutated gene to wildtype in an adult organism. You decide to use the new CRISPR system to accomplish this task. Three important CRISPR-related components must be included in the virus that will be used to treat the subject. What are these three components, and what is the function of each?

Expert Solution

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.

CRISPR relies on just two components: the molecular scissors, a CRISPR-associated (Cas) nuclease, and the GPS guiding it to the appropriate site, the guide RNA (gRNA). In natural, the gRNA consists of two distinct segments of RNA: CRISPR RNA (crRNA) and transactivating CRISPR RNA (tracrRNA).

Cas nuclease Cas9 (CRISPR associated protein 9) is a protein which plays a vital role in the immunological defense of certain bacteria against DNA viruses and plasmids and which is heavily utilized in genetic engineering applications. Its main function is to cut DNA and therefore it can alter a cell's genome.

The guide RNA is a specific RNA sequence that recognizes the target DNA region of interest and directs the Cas nuclease there for editing. ... The CRISPR-associated protein is a non-specific endonuclease.

gladiator answered 2 years ago

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