Lecture 20 Flashcards
CRISPR stands for
Clustered Regularly Interspaced Short Palindromic Repeats
How did they study by Diakatou for the allele-specific knockout try to do?
Knockout the domiant mutant allele so that the wild type can perform unhindered
CRISPR-Cas9
induce DNA breaks
The challenge of a G56R-Knockout stratege was__
to design allele-specific gRNAs that would not target the WT
Photo 25
The area highlighted gRNA1 showed Cas9 being used only in the heterozygous cell line
If Cas9 worked it would have created insertions and deletions making T7E1 cut DNA producing mismathces and extra bands
Where did CRISPR for knowkcing out alleles come from?
mechanism of adaptive immunity in bacteria
-study involving iap(bacteria) found unusual structure at the 3’ end- 29 highly conserved nucleotides
CRISPR was first called
SRSRs
short regularly spaced repeats
CRISPR are a distinct feature in bacteria for?
resistance of bacteriophages with the association of CAS
Barrangou eat al. 2007 investigated
photo 26
the origin and function of additional spacers present in phage-resistant strains.
Explain photo 26 involved with Barrangou
the number of spacers effect immunity
858 and 2972 are two different viruses
The dual____when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage>
tracrRNA:crRNA
pre-crRNA
multi-spacer array
crRNA
fragments
IN type 11 CRISPR the ___is processed differently.
pre-crRNA
PAM sequence acts like a ____.Cas9 scans dsDNA quickly for ____
BLAST word size
PAM sequences
