期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 115, 期 10, 页码 E2183-E2192出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1714530115
关键词
oligonucleotide; FISH; in situ; superresolution; oligo
资金
- National Institutes of Health Awards [1R01EB018659-01, 1-U01-MH106011-01, DP1GM106412, R01HD091797]
- Office of Naval Research Awards [N00014-13-1-0593, N00014-14-1-0610, N00014-16-1-2182, N00014-16-1-2410]
- National Science Foundation (NSF) Awards [CCF-1054898, CCF-1317291]
- Damon Runyon Cancer Research Foundation Fellowship
- Uehara Memorial Foundation Research Fellowship
- European Molecular Biology Organization
- Human Frontier Science Program
- NSF Graduate Research Fellowships
Oligonucleotide (oligo)-based FISH has emerged as an important tool for the study of chromosome organization and gene expression and has been empowered by the commercial availability of highly complex pools of oligos. However, a dedicated bioinformatic design utility has yet to be created specifically for the purpose of identifying optimal oligo FISH probe sequences on the genome-wide scale. Here, we introduce OligoMiner, a rapid and robust computational pipeline for the genome-scale design of oligo FISH probes that affords the scientist exact control over the parameters of each probe. Our streamlined method uses standard bioinformatic file formats, allowing users to seamlessly integrate new and existing utilities into the pipeline as desired, and introduces a method for evaluating the specificity of each probe molecule that connects simulated hybridization energetics to rapidly generated sequence alignments using supervised machine learning. We demonstrate the scalability of our approach by performing genome-scale probe discovery in numerous model organism genomes and showcase the performance of the resulting probes with diffraction-limited and single-molecule superresolution imaging of chromosomal and RNA targets. We anticipate that this pipeline will make the FISH probe design process much more accessible and will more broadly facilitate the design of pools of hybridization probes for a variety of applications.
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