Gene editing (Max)

Question 1

what are ZFN’s

Answer 1

artificial Restriction enzymes. Made by the fusion of ZF DNA binding domain and DNA cleavage domain

Question 2

how do ZFN’s work

Answer 2

• ZFN into cell by electroporation
  -ZF protein recognise target site
  -FOKI cut, double strand break
• gene delete or frag inserted into.

Question 3

ZFN +/-‘ve

Answer 3

+precise
- cost
-make whole system for new
-hard to modify

Question 4

what are TALENS

Answer 4

proteins from xanthomones bacteria

Question 5

How do TALEN’s work?

Answer 5

-Engineer TALEN
- into a plasmid, then plasmid into a cell
- express TALENS
- FOK1 cut and caused the double-strand break
- delete of gene alter gene

Question 6

TALENS +/-‘ve

Answer 6

+Med apps
+ precise
- hart to patent

Question 7

how does CRISPR-Cas9 work?

Answer 7

-CRISPR-Cas9 attaches to DNA and cut
- Cas9+ guide RNA
-gRNA complementary to DNA target
- Cas9 cut across both strands causing double-strand break
- DNA repair

Question 8

CRISPR-Cas9 +/-‘ve

Answer 8

+ cheap
+easy to amend target
- off-target effects
- time if first time setting up

Question 9

how is yoghurt linked to CRISPR

Answer 9

phage skill bacteria in yoghurt
archaea prev infect by phage DNA of this remain I CRISPR library
reinfect by phage- CRISPR cut phage, inactivating it.