Fluorescent in vivo editing reporter (FIVER): A novel multispectral reporter of in vivo genome editing
Peter A. Tennant,
Robert G. Foster,
Daniel O. Dodd,
Ieng Fong Sou,
Laura C Murphy,
Margaret A Keighren,
Stephen L. Hart,
Posted 14 Jul 2020
bioRxiv DOI: 10.1101/2020.07.14.200170
Posted 14 Jul 2020
Advances in genome editing technologies have created opportunities to treat rare genetic diseases, which are often overlooked in terms of therapeutic development. Nonetheless, substantial challenges remain: namely, achieving therapeutically beneficial levels and kinds of editing in the right cell type(s). Here we describe the development of FIVER (fluorescent in vivo editing reporter) - a modular toolkit for in vivo detection of genome editing with distinct fluorescent read-outs for non-homologous end-joining (NHEJ), homology-directed repair (HDR) and homology-independent targeted integration (HITI). We demonstrate that fluorescent outcomes reliably report genetic changes following editing with diverse genome editors in primary cells, organoids and in vivo . We show the potential of FIVER for high-throughput unbiased screens, from small molecule modulators of genome editing outcomes in primary cells through to genome-wide in vivo CRISPR cancer screens. Importantly, we demonstrate its in vivo application in postnatal organ systems of interest for genetic therapies - retina and liver. FIVER will broadly help expedite the development of therapeutic genome surgery for many genetic disorders. ### Competing Interest Statement The authors have declared no competing interest.
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