Detection of marker-free precision genome editing and genetic variation through the capture of genomic signatures Journal Article
| Authors: | Billon, P.; Nambiar, T. S.; Hayward, S. B.; Zafra, M. P.; Schatoff, E. M.; Oshima, K.; Dunbar, A.; Breinig, M.; Park, Y. C.; Ryu, H. S.; Tschaharganeh, D. F.; Levine, R. L.; Baer, R.; Ferrando, A.; Dow, L. E.; Ciccia, A. |
| Article Title: | Detection of marker-free precision genome editing and genetic variation through the capture of genomic signatures |
| Abstract: | Genome editing technologies have transformed our ability to engineer desired genomic changes within living systems. However, detecting precise genomic modifications often requires sophisticated, expensive, and time-consuming experimental approaches. Here, we describe DTECT (Dinucleotide signaTurE CapTure), a rapid and versatile detection method that relies on the capture of targeted dinucleotide signatures resulting from the digestion of genomic DNA amplicons by the type IIS restriction enzyme AcuI. DTECT enables the accurate quantification of marker-free precision genome editing events introduced by CRISPR-dependent homology-directed repair, base editing, or prime editing in various biological systems, such as mammalian cell lines, organoids, and tissues. Furthermore, DTECT allows the identification of oncogenic mutations in cancer mouse models, patient-derived xenografts, and human cancer patient samples. The ease, speed, and cost efficiency by which DTECT identifies genomic signatures should facilitate the generation of marker-free cellular and animal models of human disease and expedite the detection of human pathogenic variants. © 2020 The Author(s) Billon et al. report the development of a versatile detection method based on the capture of targeted genomic signatures. This method allows the detection and quantification of genomic signatures introduced by marker-free precision genome editing or resulting from genetic variation. © 2020 The Author(s) |
| Keywords: | controlled study; unclassified drug; gene mutation; human cell; nonhuman; animal cell; mouse; animal tissue; analytic method; animal experiment; animal model; gene frequency; genetic variation; tumor xenograft; mutational analysis; cancer model; amplicon; restriction endonuclease; sequence homology; genetic marker; dinucleotide; genomic dna; mammal cell; homology-directed repair; detection method; measurement accuracy; human; male; female; priority journal; article; crispr; clustered regularly interspaced short palindromic repeat; organoid; gene editing; base editing; dinucleotide signatures; human pathogenic variants; precision genome editing; prime editing; type iis restriction endonucleases; acui protein; dinucleotide signature capture method |
| Journal Title: | Cell Reports |
| Volume: | 30 |
| Issue: | 10 |
| ISSN: | 2211-1247 |
| Publisher: | Cell Press |
| Date Published: | 2020-03-10 |
| Start Page: | 3280 |
| End Page: | 3295.e6 |
| Language: | English |
| DOI: | 10.1016/j.celrep.2020.02.068 |
| PUBMED: | 32160537 |
| PROVIDER: | scopus |
| PMCID: | PMC7108696 |
| DOI/URL: | |
| Notes: | Article -- Source: Scopus |
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