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- W4319602705 abstract "ABSTRACT Generating high-coverage sequencing coverage at select genomic loci has extensive applications in both research science and genetic medicine. Long-read sequencing technologies (e.g. nanopore sequencing) have expanded our ability to generate sequencing data in regions (e.g. repetitive elements) that are difficult to interrogate with short-read sequencing methods. In work presented here, we expand on our previous work using CRISPR/Cas9 for targeted nanopore sequencing by using in vitro transcribed guideRNAs, with 1100 guideRNAs in a single experiment. This approach decreases the cost per guideRNA, increases the number of guideRNAs that can be multiplexed in a single experiment, and provides a way to rapidly screen numerous guideRNAs for cutting efficiency. We apply this strategy in multiple patient-derived pancreatic cancer cell lines, demonstrating its ability to unveil structural variation in “deletion hotspots” around the tumor suppressor genes p16 ( CDKN2A ), and SMAD4 ." @default.
- W4319602705 created "2023-02-09" @default.
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- W4319602705 date "2023-02-07" @default.
- W4319602705 modified "2023-10-03" @default.
- W4319602705 title "IVT generation of guideRNAs for Cas9-enrichment Nanopore Sequencing" @default.
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- W4319602705 doi "https://doi.org/10.1101/2023.02.07.527484" @default.
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