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mRNA - Coggle Diagram
mRNA
shRNA pros
long term knockdown
delivery to virtually any cell line
renewable
can culture in lab to get more
in vivo use
CRISPR
Mechanism
1) gRNA scaffold binds to the CAS protein
2) binds to DNA, unwinds and makes double stranded break
3) needs to be prone to NHEJ error
CRISPR RNA
tefers to target specific sequence
CRISPR CAS 9 PAM
protection against self cleavage
Other applications
base editing
dead Cas9
curing sickle cell disease
caused by small amino acid change
modified blood with crispr Cas 9
cons of siRNA
nonrenewable and only transient
challenging in vivo
shRNA cons
needs cloning
can be technically challenging
Viral transduction
results in stable transfection
virus is able to give DNA and RNA materials
RNA interference
the cell can degrade foreign dsRNA molecules
consequence in transfection: knocks down gene of interest
pros of siRNA
easily titrated (fast and simple)
CRISPR like tool
researchers ...
nanoparticle siRNA transfection