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The Cas9 is a protein enzyme that cuts DNA at targets specified by a short guide RNA bound to it. The sgRNA includes a 20-nucleotide sequence that is complementary to the DNA strand that it will cut. If the target DNA site has to be perfectly complementary to all 20 nucleotides, approximately how many sites will be cut in the human genome for a particular sgRNA? The human genome is 3 billion (3 x 10E9) nucleotides per haploid set of chromosomes.In other, very pertinent terms for human gene therapy, if you design an sgRNA to cut at a particular site in a human gene, how many off-target sites do you think the sgRNA-Cas9 complex will cut in the rest of the genome? (Set up and solve the problem with your group.)

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