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Where is the PAM site (NGG) located? (1pt) A. In the gRNA spacer B. Downstream (towards...

Where is the PAM site (NGG) located? (1pt) A. In the gRNA spacer B. Downstream (towards the 3’ end) of the target genomic strand that is complementary to the gRNA spacer sequence C. Upstream (towards the 5’ end) of the target genomic strand that is homologous to the gRNA spacer sequence D. In the gRNA scaffold E. Downstream (towards the 3’ end) of the target genomic strand that is homologous to the gRNA spacer sequence F. In the pMD2-CAN1.Y-gRNA plasmid

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Answer #1

PAM site (NGG) is located on the downstream (towards 3' end) of the target genomic strand that is homologous to the gRNA spacer sequence .

Interaction between guanines of PAM and the Arginines of S. pyogenes Cas 9 , positions the phosphate of the DNA backbone 5' to the PAM to interact with a phosphate - lock loop in Cas9 and facilitate DNA strand unwinding.If DNA is complimentary to the gRNA ,an RNA: DNA hybrid forms, called an R loop and cleavage follows.

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