ArticleChemical communications (Cambridge, England)2020
Messenger RNA enrichment using synthetic oligo(T) click nucleic acids.
Article in Chemical communications (Cambridge, England), 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
5 citing papers in PubMed, 16 citations in OpenAlex.
- The Regulation of Ferroptosis by Noncoding RNAs.International journal of molecular sciences · 2023Review
- Epitranscriptomics in parasitic protists: Role of RNA chemical modifications in posttranscriptional gene regulation.PLoS pathogens · 2022Review
- Identification of Key Genes during Ethylene-Induced Adventitious Root Development in Cucumber (International journal of molecular sciences · 2022Article
- Circular RNA: A promising new star for the diagnosis and treatment of colorectal cancer.Cancer medicine · 2021Review
- Review
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Authors and funding
7 authors at 1 institution in 2 countries.
Funding
Abstract
Enrichment of mRNA is a key step in a number of molecular biology techniques, particularly in the rapidly growing field of transcriptomics. Currently, mRNA is isolated using oligo(thymine) DNA (oligo(dT)) immobilized on solid supports, which binds to the poly(A) tail of mRNA to pull the mRNA out of solution through the use of magnets or centrifugal filters. Here, a simple method to isolate mRNA by complexing it with synthetic click nucleic acids (CNAs) is described. Oligo(T) CNA bound efficiently to mRNA, and because of the insolubility of CNA in water, >90% of mRNA was readily removed from solution using this method. Simple washing, buffer exchange, and heating steps enabled mRNA's enrichment from total RNA, with a yield of 3.1 ± 1.5% of the input total RNA by mass, comparable to the yield from commercially available mRNA enrichment beads. Further, the integrity and activity of mRNA after CNA-facilitated pulldown and release was evaluated through two assays. In vitro translation of EGFP mRNA confirmed the translatability of mRNA into functional protein and RT-qPCR was used to amplify enriched mRNA from total RNA extracts and compare gene expression to results obtained using commercially available products.
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Registered trials
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