Discuss the following questions;
Questions
1. What differences between prokaryotic and eukaryotic cells are important to consider when adapting Cas9 for eukaryotic gene editing?
2. What components need to be introduced to make a Cas9 induced break in eukaryotic cells?
3. Are Cas9 proteins found in humans? If so, what is their role? If not, why not?
Questions
1. How would you determine and what strategies could you use to ensure that no off-target background mutations occurred when editing with Cas9?
2. What advantages and disadvantages are there to each of the different ways of introducing Cas9 and sgRNA?
3. What type of desired mutations require a donor template to also be injected in addition to the sgRNA and Cas9?
Questions
1. What are other uses you could think of for the dCas9 protein?
2. What are the differences in repressing a gene with CRISPRi versus inactivating a gene product by making a mutation with CRISPR gene editing?
3. What are possible limitations of Cas9 targeting methods?
