Gene Editing Flashcards
What does a CRISPR/Cas9 plasmid contain?
Promotor, guide RNA, CRISPR RNA, another promotor, CAS9 gene, fluorescence gene.
How is the bacteria CAS9 modified to engineering use?
Added nuclear signalling sequence.
What must follow the DNA sequence targeted by CRISPR/Cas9?
Protospacer adjacent motif (PAM).
What is the protospacer adjacent motif?
Any nucleotide followed by GG.
What is the size of the DNA sequence recognised by a pair of Z-fingers, TALENs and CRISPR/Cas9?
18-36, 28-40, 22, respectively.
What are the trageting Z-fingers, TALENs and CRISPR/Cas9?
Z-fingers: Difficult to target non-G-rich sequences
TALENs: 5’ targeted base must be T
CRISPR: PAM site
List Z-fingers, TALENs and CRISPR/Cas9 in decreasing order of difficulty to engineer.
Z-fingers, TALENs and CRISPR/Cas9.
How can Z-fingers, TALENs and CRISPR/Cas9 be delivered ex-vivo?
Electroporation and viral transduction.
List three disease that can be treated with Z-fingers.
Haemophilia B, HIV, SCID
List three diseases that can be treated with TALENs
DMD, Hep B, B-cell accute lymphoblastic leukaemia.
List seven disease that can potentially be treated with CRISPR/Cas9.
HIV, DMD, cataracts, cystic fibrosis, hereditary tyrosinemia.