Stockejoztci

Once the PAM is recognized, the guide region of the gRNA undergoes seed nucleation to form an Aformlike helical RNADNA hybrid duplex Only once the RNA and DNA complete Rloop formation, also known as the zipped conformation, and structural rearrangement of the nuclease domains commence, can the endonuclease cut the DNA creating a DSB ( Jiang et al, 15 ;In addition, the offtarget effect of an engineered gRNACas9 was found on an imperfectly paired genomic site, but it had lower genomeediting efficiency than the perfectly matched site Further analysis suggests that mismatch position between gRNA seed and target DNA is an important determinant of the gRNACas9 targeting specificity, andThe 5′end of gRNA (gRNA seed) is shown to pair with one strand of targeted DNA The scaffold of gRNA is labeled with darkred cycles A PAM motif (NGG) is located adjacent to the DNA– gRNA pairing region in the complementary strand of targeted DNA The DNA–gRNA basepairing should be at least 15bp long The Cas9 nuc...