What are the most promising (potential) uses of the CRISPR technology?

The CRISPR-Cas9 complex has revolutionized genomic research because of its very consistent approach for identifying and cutting specific sections of DNA. This complex is made from a Cas9 protein which cuts DNA through two if its nuclease domains, and single guide RNA (sgRNA) which targets a specific section of the DNA called a PAM.

Although its ability to make double-stranded breaks in DNA is remarkable, because it allows scientists to study knockout genes, people have begun to envision new uses for CRISPR. Most of these new uses stem from the Cas9-complex’s ability to move to certain spots in a DNA sequence consistently due to the sgRNA.

One way scientists have exploited this specific movement is by eliminating the Cas9 protein’s ability to cut DNA and fusing other enzymes to it. Now, when the Cas9 complex reaches a specific sequence of DNA, the fused enzyme will perform a specific function on it rather than Cas9 cutting the DNA. One such example is fusing deaminases onto Cas9 such that it can mutate specific bases in the DNA, like replacing a Cytosine base with a Thymine. As you can tell, this idea comes with immense power, because through these mutations we can delete deadly genetic diseases and alter premature stop codons.

CRISPR can also be used to modulate gene expression, like increasing transcription, by fusing “transcription activators” onto Cas9. This allows Cas9 to transport transcription activators to a specific gene and then attract the other transcription enzymes found in the cell such as DNA polymerase and RNA primase. Or it could silence the gene by recruiting other proteins which will bind to the DNA and physically block transcription.

Yet another use of CRISPR is to attach fluorescent proteins onto the Cas9 complex so that scientists can visualize where specific sections of DNA are found in the cell.

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