2.5: Crispr

Question 1

What does CRISPR stand for? What is it?

Answer 1

Clustered regularly interspaced short palindromic repeats.
Organization of a viral DNA segment into the bacterial genome, so the bacteria makes a DNA copy of the viral genome and incorporates it into its genome.

Question 2

1 step of CRISPR. What are the 2 components involved?

Answer 2

Guide RNA, engineered to be complementary base paring to the target DNA.
Cas9 protein, cleaves the target DNA when paired to the guide RNA
1st step: Transform a cell with plasmid that contains both guide RNA and Cas9. Guide RNA undergoes shape change to a hairpin structure.

Question 3

2 step of CRISPR. What is the new component involved?

Answer 3

Exonuclease

2nd step: Cas9 cleaves the target DNA when guide RNA undergoes base pairing. Exonuclease expands the gap.

Question 4

3 step of CRISPR. What is the final component?

Answer 4

Editing template DNA
Last step: Editing template DNA contains the sequence of interest that will replace the cleaved target DNA. Strands of the gapped target DNA undergo base pairing with the editing template and DNA synthesis closes the gap.