CRISPR

Question 1

what is the goal of genetic engineering?

Answer 1

to alter the DNA sequence of cells, or an organism in a precise manner in order to change its characteristics

Question 2

restirction enzymes

Answer 2

bacterial defense against viral infection by restriction-modification complexes
-looks for specific viral DNA sequence and cleaves it
-cut double stranded DNA, leaving sticky ends creating recombinant DNA

Question 3

restriction enzymes+plasmids+DNA ligase

Answer 3

-human DNA and bacterial DNA are cut using restriction enzymes and fused, making recombinant DNA
-DNA ligase seals the nicks

Question 4

what was issues with the old idea of gene therapy

Answer 4

-add new, good copy of the gene into someone who has a bad copy
-doesn’t remove the old gene, but the new gene should off set the effects of the old one
-can’t control where in the chromosome the gene is added, resulting in serious side affects

Question 5

what are methods to get site-specific DNA editing?

Answer 5

-zinc finger nucleases (lot of time and money)
-TALENS (lot of time and money)
-CRISPR/Cas9 (quick and inexpensive)

Question 6

CRISPR/Cas9

Answer 6

R=repeat
S=spacers (previous viral invaders that associate with Cas9)
-Cas9 is a bacterial protein that binds to the “repeat” sequence of each RNA (forming a complex)
-“sentinal”=complex that looks for viral DNA that matches the “spacer” part of its RNA
-when it finds a match it binds and cuts it

Question 7

how can CRISPR be utilized to recognize our own gene of interest?

Answer 7

1. Cas9: does DNA cutting
2. guide RNA to target Cas9/RNA complex to desired strech of DNA
3. donor DNA fragment to match broken ends of DNA and provide new sequence for desired alteration in the DNA sequence

Question 8

CRISPR

Answer 8

clustered regulated interspaced short palindromic repeats