Abstract: The present disclosure provides genetically modified cells and non-human organisms, and methods of producing such cells and organisms. Also provided are methods of editing the genome of such cells and organisms. The cells and non-human organisms of the disclosure are genetically modified such that their genome includes an integrated heterologous nucleic acid (that includes a CRISPR/Cas target sequence) at one or more (e.g., 2 or more, 3 or more, 4 or more, etc.) positions within the genome. The integrated nucleic acids, which include the same CRISPR/Cas target sequence, allow for simultaneous gene editing at multiple different positions within a genome using a single species of CRISPR/Cas guide RNA (i.e., one guide RNA will target multiple sites because the multiple sites have the same target sequence).

Abstract: The present disclosure provides genetically modified cells and non-human organisms, and methods of producing such cells and organisms. Also provided are methods of editing the genome of such cells and organisms. The cells and non-human organisms of the disclosure are genetically modified such that their genome includes an integrated heterologous nucleic acid (that includes a CRISPR/Cas target sequence) at one or more (e.g., 2 or more, 3 or more, 4 or more, etc.) positions within the genome. The integrated nucleic acids, which include the same CRISPR/Cas target sequence, allow for simultaneous gene editing at multiple different positions within a genome using a single species of CRISPR/Cas guide RNA (i.e., one guide RNA will target multiple sites because the multiple sites have the same target sequence).