LECTURE 13 Flashcards
genome editing
type of genetic engineering in which DNA is inserted, deleted, modified or replaced using nucleases
what is CRISPR?
Clustered Regularly Interspaced Short Palindromic Repeats
what is the origin of CRISPR
it is an adaptive immune repose in prokaryotes
How does CRISPR work in prokaryotes?
invading DNA is recognised and cut by Cas1-Cas2 protein complexes into protospacers
protospacers integrate into the CRISPR locus in the bacterial genome
upon reinfection transcription of the protospacers to RNA is activated which binds to Cas9
Cas9 cuts DNA strands to prevent infection
what is the name given to the transactvathng RNA and protospacer complex
guide RNA
which part of the CRISPR locus encodes Cas proteins
cas operon
what is PAM?
Protospacer Adjacent Motif, necessary sequence found adjacent to the protospacer in the target DNA, discriminates target from self
Homology directed repair (HDR)
an error prone repair mechanism that introduces insertions/deletions into the DNA impacting gene function
key to CRISPR knockout studies
Non-homologous end joining (NHEJ)
DNA repair mechanism that precisely repairs DNA using sister chromatids during S phase
key to CRISPR knock in studies
current androgen treatment for prostate cancer
inactivate AR by blocking ligand binding
Ex-Vivo delivery of CRISPR
remove cells from patient
edit genome
screen
engraft cells back into patient
In vivo delivery of CRISPR
package CRISPR in a delivery vehicle
deliver to patient
