دورية أكاديمية
أعرض في EDS

A single-copy knockin translating ribosome immunoprecipitation toolkit for tissue-specific profiling of actively translated mRNAs in C . elegans .

التفاصيل البيبلوغرافية
العنوان: A single-copy knockin translating ribosome immunoprecipitation toolkit for tissue-specific profiling of actively translated mRNAs in C . elegans .
المؤلفون: Wester LE; Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA.; Department of Molecular Genetics of Ageing, Max Planck Institute for Biology of Ageing, 50668 Cologne, Germany., Lanjuin A; Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA., Bruckisch EHW; Department of Molecular Genetics of Ageing, Max Planck Institute for Biology of Ageing, 50668 Cologne, Germany., Perez-Matos MC; Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA., Stine PG; Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA., Heintz C; Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA., Denzel MS; Department of Molecular Genetics of Ageing, Max Planck Institute for Biology of Ageing, 50668 Cologne, Germany.; Altos Labs, Cambridge, UK., Mair WB; Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA.
المصدر: Cell reports methods [Cell Rep Methods] 2023 Mar 27; Vol. 3 (3), pp. 100433. Date of Electronic Publication: 2023 Mar 27 (Print Publication: 2023).
نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Elsevier Inc Country of Publication: United States NLM ID: 9918227360606676 Publication Model: eCollection Cited Medium: Internet ISSN: 2667-2375 (Electronic) Linking ISSN: 26672375 NLM ISO Abbreviation: Cell Rep Methods
أسماء مطبوعة: Original Publication: [New York] : Elsevier Inc., [2021]-
مواضيع طبية MeSH: Caenorhabditis elegans*/genetics , Protein Biosynthesis*/genetics, Animals ; RNA, Messenger/genetics ; Ribosomes/genetics ; Ribosomal Proteins/genetics ; Immunoprecipitation
مستخلص: Here, we introduce a single-copy knockin translating ribosome immunoprecipitation (SKI TRIP) toolkit, a collection of Caenorhabditis elegans strains engineered by CRISPR in which tissue-specific expression of FLAG-tagged ribosomal subunit protein RPL-22 is driven by cassettes present in single copy from defined sites in the genome. Through in-depth characterization of the effects of the FLAG tag in animals in which endogenous RPL-22 has been tagged, we show that it incorporates into actively translating ribosomes and efficiently and cleanly pulls down cell-type-specific transcripts. Importantly, the presence of the tag does not impact overall mRNA translation, create bias in transcript use, or cause changes to fitness of the animal. We propose SKI TRIP use for the study of tissue-specific differences in translation and for investigating processes that are acutely sensitive to changes in translation like development or aging.
Competing Interests: The authors declare no competing interests.
(© 2023 The Author(s).)
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معلومات مُعتمدة: R01 AG044346 United States AG NIA NIH HHS; R01 AG051954 United States AG NIA NIH HHS; R01 AG059595 United States AG NIA NIH HHS; R01 AG067106 United States AG NIA NIH HHS
فهرسة مساهمة: Keywords: C. elegans; RPL-22; SKI TRIP; TRAP; ribotag; translation; translatome
المشرفين على المادة: 0 (RNA, Messenger)
0 (Ribosomal Proteins)
تواريخ الأحداث: Date Created: 20230414 Date Completed: 20231020 Latest Revision: 20240210
رمز التحديث: 20240210
مُعرف محوري في PubMed: PMC10088236
DOI: 10.1016/j.crmeth.2023.100433
PMID: 37056370
قاعدة البيانات: MEDLINE
الوصف
تدمد:2667-2375
DOI:10.1016/j.crmeth.2023.100433