Abstract
Over the past decade, modifications to microRNAs (miRNAs) via 3' end nucleotide addition have gone from a deep-sequencing curiosity to experimentally confirmed drivers of a range of regulatory activities. Here we overview the methods that have been deployed by researchers seeking to untangle these diverse functional roles and include characterizing not only the nucleotidyl transferases catalyzing the additions but also the nucleotides being added, and the timing of their addition during the miRNA pathway. These methods and their further development are key to clarifying the diverse and sometimes contradictory functional findings presently attributed to these nucleotide additions.
Highlights
The development and subsequent adoption of high-throughput sequencing as a tool to capture snapshots of the cellular transcriptome have considerably sharpened the collective resolution of cellular RNA landscapes
This review describes current methods for identification of 3’ terminal nucleotide additions through both experimental and computational approaches
We focus on discussing the current methods for isolating and deepsequencing cellular pre-miRNA populations, given that 3’ nucleotide addition to pre-miRNAs has distinct functional implications (Figure 1)
Summary
The development and subsequent adoption of high-throughput sequencing as a tool to capture snapshots of the cellular transcriptome have considerably sharpened the collective resolution of cellular RNA landscapes. Deep sequencing technologies provide a top-down approach to identifying potential miRNA loci that are significantly influenced by terminal nucleotide addition events.
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