دورية أكاديمية
أعرض في EDS

Analysis of C. elegans Germline Small RNA Pathways.

التفاصيل البيبلوغرافية
العنوان: Analysis of C. elegans Germline Small RNA Pathways.
المؤلفون: Renaud MS; Department of Molecular Genetics, University of Toronto, Toronto, ON, USA., Seroussi U; Department of Molecular Genetics, University of Toronto, Toronto, ON, USA., Claycomb JM; Department of Molecular Genetics, University of Toronto, Toronto, ON, USA. julie.claycomb@utoronto.ca.
المصدر: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2677, pp. 37-59.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE
أسماء مطبوعة: Publication: Totowa, NJ : Humana Press
Original Publication: Clifton, N.J. : Humana Press,
مواضيع طبية MeSH: MicroRNAs*/genetics , RNA, Small Untranslated*/genetics, Animals ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans/metabolism ; Germ Cells/metabolism ; Gene Expression Regulation ; Argonaute Proteins/genetics ; Argonaute Proteins/metabolism
مستخلص: Sequence-specific gene regulation by small RNA (sRNA) pathways is essential for the development and function of organisms in all domains of life. These regulatory complexes, containing an Argonaute protein (AGO) guided by a bound sRNA, have the potential to regulate thousands of individual target transcripts at both the co- and post-transcriptional level. Determining the repertoire of transcripts that an AGO is capable of regulating in a particular context is essential to understanding the function of these regulatory modules. Immunoprecipitation (IP) of AGOs and subsequent RNA sequencing of their bound sRNAs allows for the inference of their target transcripts by mapping the sequences of the co-precipitated sRNAs back to their complementary target transcripts. This approach can be complemented by sequencing sRNAs from ago mutants as sRNA transcripts are degraded in the absence of their AGO binding partner. Here, we describe a framework for analyzing AGO/sRNA pathways in the germline, from using CRISPR-Cas9 to tag or mutate AGOs, through protocols for the extraction, sequencing, and analysis of sRNAs from AGO IPs and ago mutants.
(© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
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فهرسة مساهمة: Keywords: Argonaute; C. elegans; CRISPR; Immunoprecipitation; RNA-seq; miRNA; piRNA; sRNA; siRNA
المشرفين على المادة: 0 (MicroRNAs)
0 (Argonaute Proteins)
0 (RNA, Small Untranslated)
تواريخ الأحداث: Date Created: 20230718 Date Completed: 20230721 Latest Revision: 20230721
رمز التحديث: 20230721
DOI: 10.1007/978-1-0716-3259-8_2
PMID: 37464234
قاعدة البيانات: MEDLINE
الوصف
تدمد:1940-6029
DOI:10.1007/978-1-0716-3259-8_2