AI Article Synopsis
- RNA sequencing analysis requires isolating protein-coding transcripts, which can be done by enriching desired coding fractions or removing abundant non-coding rRNA.
- The study introduces a new method using the antiviral protein IFIT1, which effectively captures mRNA while depleting rRNA, providing high-quality RNA-seq data.
- IFIT1 is presented as a cost-effective and versatile tool for preparing mRNA libraries across various organisms with cap 0 mRNA ends, including plants and fungi.
Article Abstract
Analysis of the protein coding transcriptome by the RNA sequencing requires either enrichment of the desired fraction of coding transcripts or depletion of the abundant non-coding fraction consisting mainly of rRNA. We propose an alternative mRNA enrichment strategy based on the RNA-binding properties of the human IFIT1, an antiviral protein recognizing cap 0 RNA. Here, we compare for Saccharomyces cerevisiae an IFIT1-based mRNA pull-down with yeast targeted rRNA depletion by the RiboMinus method. IFIT1-based RNA capture depletes rRNA more effectively, producing high quality RNA-seq data with an excellent coverage of the protein coding transcriptome, while depleting cap-less transcripts such as mitochondrial or some non-coding RNAs. We propose IFIT1 as a cost effective and versatile tool to prepare mRNA libraries for a variety of organisms with cap 0 mRNA ends, including diverse plants, fungi and eukaryotic microbes.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9825183 | PMC |
| http://dx.doi.org/10.1093/nar/gkac903 | DOI Listing |
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