دورية أكاديمية
أعرض في EDS

CRISPR-Cas9-mediated host signal reduction for 18S metabarcoding of host-associated eukaryotes.

التفاصيل البيبلوغرافية
العنوان: CRISPR-Cas9-mediated host signal reduction for 18S metabarcoding of host-associated eukaryotes.
المؤلفون: Owens LA; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA., Thurber MI; Henry Vilas Zoo, Madison, Wisconsin, USA., Goldberg TL; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA.
المصدر: Molecular ecology resources [Mol Ecol Resour] 2024 Aug; Vol. 24 (6), pp. e13980. Date of Electronic Publication: 2024 May 28.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Blackwell Country of Publication: England NLM ID: 101465604 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1755-0998 (Electronic) Linking ISSN: 1755098X NLM ISO Abbreviation: Mol Ecol Resour Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Oxford, England : Blackwell
مواضيع طبية MeSH: CRISPR-Cas Systems* , Eukaryota*/genetics , DNA Barcoding, Taxonomic*/methods , RNA, Ribosomal, 18S*/genetics, Metagenomics/methods ; Humans ; Animals
مستخلص: Metabarcoding-based methods for identification of host-associated eukaryotes have the potential to revolutionize parasitology and microbial ecology, yet significant technical challenges remain. In particular, highly abundant host reads can mask the presence of less-abundant target organisms, especially for sample types rich in host DNA (e.g., blood and tissues). Here, we present a new CRISPR-Cas9-mediated approach designed to reduce host signal by selective amplicon digestion, thus enriching clinical samples for eukaryotic endosymbiont sequences during metabarcoding. Our method achieves a nearly 76% increased efficiency in host signal reduction compared with no treatment and a nearly 60% increased efficiency in host signal reduction compared with the most commonly used published method. Furthermore, the application of our method to clinical samples allows for the detection of parasite infections that would otherwise have been missed.
(© 2024 The Author(s). Molecular Ecology Resources published by John Wiley & Sons Ltd.)
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معلومات مُعتمدة: R01 AG049395 United States AG NIA NIH HHS; T32 AI007414 United States AI NIAID NIH HHS; 1R21AI163592-01 United States HI NHLBI NIH HHS; T32AI007414 United States HI NHLBI NIH HHS; 1R01AG049395-01 United States HI NHLBI NIH HHS; R21 AI163592 United States AI NIAID NIH HHS
فهرسة مساهمة: Keywords: CRISPR‐Cas9; eukaryotic microbiome; metabarcoding; parasitology
المشرفين على المادة: 0 (RNA, Ribosomal, 18S)
تواريخ الأحداث: Date Created: 20240528 Date Completed: 20240730 Latest Revision: 20240802
رمز التحديث: 20240802
مُعرف محوري في PubMed: PMC11288772
DOI: 10.1111/1755-0998.13980
PMID: 38804043
قاعدة البيانات: MEDLINE
الوصف
تدمد:1755-0998
DOI:10.1111/1755-0998.13980