CRISPR-Cas9 has made genetic engineering easier, faster, and cheaper than ever before. A scientist interested in manipulating a particular gene only needs to search the gene’s sequence for a suitable PAM. Once a PAM is found, the corresponding Cas9 can be ordered or harvested from its bacterial strain (and as I mentioned last week, even if a PAM isn’t found, it is possible to engineer a Cas9 to recognize a new PAM sequence). An appropriate sgRNA (the crRNA:tracrRNA fusion molecule) can be designed by identifying the target sequence 20 nucleotides upstream of the chosen PAM. These sgRNA’s can be engineered…
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