The cas9 protein
has gained major attraction in the recent years simply because it can cleave
nearly any sequence complementary to the guide RNA.It is majorly associated
with the CRISPR which is an adaptive immunity system among other bacteria. The
interaction of Cas9 protein with target DNA is always very tight that the high
morality urea protein denaturant cannot usually fully dissociate the cas9
RNA-protein complex from DNA target. The ability of the PNA Monomer not to
degrade by either nucleases or proteases has made the scientist to investigate.
The CRISP cas9 has over the years expanded the major toolbox for
mammalian genetics which has enabled the rapid generation of isogenic cell
lines and mice modified alleles. Through the recent advances in genome
engineering technologies mainly associated with the CRISP cas9 have empowered researchers to be able to elucidate the
major functional organization of the genome at the systems level and establish
causal linkages between genetic variations.
Crispr
/cas9/pan protocol
majorly describes how to transect cultured cells with the unique CRISPR
machinery using some conventional lipofection reagents. The use of the CRISPR/cas9/ pna system for genome editing has been a major breakthrough making
the modification in cells or organism fast, robust and much more efficient that
the previous genome editing methods. The Cas9 always cleaves the target DNA
hence making it available for repair by the non-homologous end joining.
PNA
monomer is some
olingonucleotide analogues in which the standard sugar phosphate which is the
backbone for DNA is substituted by a pseudo peptide backbone. It was invented
by a group of scientist the famous university of Copenhagen. They are a hybrid
of a strongly and sequence specifically complementary DNA OR RNA. The beauty of
PNA monomer is that it is not degraded by nucleases or proteases.
These amazing properties have practically made the PNA monomer a subject of ever ending
scientific interest and the synthetic PNA Oligomers have numerous identified
and proposed applications. The group of Bhoc normally provides sufficient
protection during synthesis and is readily removed under the cleavage
conditions and often renders solubility to the monomers.
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