Lecture 1: Intro to CRISPR Flashcards
what does CRISPR stand for
clustered regularly interspaced short palindromic repeat
what is the main objective of CRISPR
it is a bacterial immune response system leveraged for genome editing
what promotes the gene editing
double-strand break repair
what are the 2 mechanisms for gene editing
non homologous end joining (NHEJ)
homology directed repair (HDR)
non homologous end joining
- mutations
- insertions and deletions
- gene knockout
(insertion and deletion mutations)
homology directed repair
- gene knockout
- gene tagging
- specific mutations/insertions/deletions
- knock-in (reporter genes)
- promoter study
(giving the cell a repair template with maybe a slight mutation here and there)
what is the guide RNA and the CRISPR RNA
the transcribed region of the unique “spacer” sequences found in CRISPR regions
what does the guide RNA and CRISPR RNA do
the transcribed spacer guides the Cas proteins to foreign genetic elements contained in the viral DNA genome
usually how long is the guide RNA for CRISPR
20 nt, and corresponds to sequences within the target gene
usually how long is the guide RNA for CRISPR
20 nt, and corresponds to sequences within the target gene
Protospacer Adjacent Motif (PAM)
a specific DNA sequence that must follow the target DNA sequence in order for Cas9 to bind and cut DNA
what is the PAM sequence for this lab, and the Cas9 protein
NGG, protein is streptococcus pyogenes
what is the central dogma
DNA => transcription => RNA => translation => protein (amino acid)
How do you figure out how DNA works?
you mutate what you want to know about, and see what happens or doesn’t happen when its messed up
what is guide rna made up of
crispr rna + tracr rna
