Recently there has been an explosion of scientist using the CRISPR-Cas9 system to make null alleles (“knockouts”) of genes.

In the past (before 2012), knocking out a gene was difficult but CRISPR-Cas9 has made the process amazingly easy. When you read about the
smallest genome a scientist stated that Venter’s study was not worth it given the power of the CRISPR-Cas9. Check out what all the buzz is
about! The two websites listed below are useful to start answering the questions below.

http://www.abc.net.au/news/2016-04-07/crispr-gene-editing-technology-explainer/7217782

http://www.yourgenome.org/facts/what-is-crispr-cas9

1) Please provide a definition for CRISPR-Cas9.

2) What is the advantage of this system of gene editing compared to other methods?

3) In what organisms was CRISPR-Cas9 first identified?

4) Use the questions below to describe how CRISPR-Cas9 can be used to inhibit a gene. Please provide a picture showing the process.

a) What is a guide RNA (gRNA)?

b) What allows this RNA to bind to the target genes?

c) What type of protein is Cas9 (what type of enzyme)

d) How does Cas-9 find its target gene?

e) What happens when Cas-9 finds the target gene?

f) What role does the DNA repair mechanism have in ensuring the target gene has been disabled?

5) What are three possible applications of CRIPR-Cas9?

6) It has been shown in human cells that with the appropriate gRNA CRISPR can knockout HIV infection from T-cells. This is an amazing
discovery. Why is it currently not possible to apply this information to humans?
7) What gene would you be interested in knocking out and from which organism?


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