In the minimally active chimeric RNA molecule that guides Cas9 DNA cleavage, one is called crRNA. I…

In the minimally active chimeric RNA molecule that guides Cas9 DNA cleavage, one is called crRNA. It is derived from two RNAs of the wild type Cas9 system: one is called tracrRNA and the other is called sgRNA. The tracrRNA is needed to activate Cas9. Based on your reading of this paper, describe the sequence components of the chimeric RNA. Select all answers that apply: (a) A 20 nucleotide guide sequence that is complementary to the target sequence in the DNA. (b) A short signal sequence derived from ribosomal RNAs. (c) A CRISPR repeat from the crRNA that occupies an internal position. (d) Sequences derived from the tracrRNA are found at the 3 #x27; end. At the target site in the DNA, the following sequences must be present: (a) A 20 bp target sequence complementary to the guide sequence in the sgRNA. (b) A short palindromic sequence called ATT, with the sequence ATTNNAAT. (c) A 3-bp protospacer motif (PAM) with the sequence NGG on one strand, adjacent to the target sequence. (d) A long palindromic sequence that readily forms a cruciform structure in supercoiled DNA. The nuclease active site that catalyzes the cleavage of each strand of the target DNA is located in Cas9. There are two identical nuclease active sites in Cas9, and each cleaves one DNA strand in the target.

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