This protocol uses CRISPR-Cas9 to create knock-out or knock-in lines. The main advantage of this technique is that it is simple and efficient to follow especially for novice researchers. For cell passaging, treat overnight cultured cells with two
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CRISPR-Cas is a powerful technology to engineer the complex genomes of plants and animals. Here, we detail a protocol to efficiently edit the human genome using different Cas endonucleases. We highlight important considerations and design parameters to optimize editing efficiency.