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Biology subjects

Sagredo, E. A.

Publications and source records attributed to Sagredo, E. A..

2 recordsLinked to original sources

ADAR1 regulates alternative splicing through an RNA editing-independent mechanism.

Adenosine-to-inosine (A-to-I) RNA editing and alternative splicing occur co-transcriptionally and can regulate and influence each other. The RNA editing enzymes ADAR1 and ADAR2 catalyze A-to-I RNA editing, where it has been shown that expression changes in both RNA editing enzymes exert alternative splicing. ADAR1 manipulation has a significant impact on alternative splicing. While many of those changes are related to changes in their A-to-I RNA editing activity, we speculate that ADAR1 may also influence splicing in an editing-independent manner. In this work, the protein-protein interactome of ADAR1 revealed that ADAR1 interacts with spliceosome co-factors and auxiliary splicing regulators. We confirmed that ADAR1 does not only influence splicing through editing but also throughout editing-independent functions with even greater penetrance. We show that ADAR1 modulates the splicing of transcripts encoding splicing factors, with many of these splice sites overlapping with known splicing changes induced by the splicing factor ACIN1, suggesting indirect effects upon ADAR1 expression. In summary, we show that ADAR1 can regulate splicing in an editing-independent manner, which likely occurs by widespread alteration of the splicing factor isoform landscape.

molecular biology↗

agoTRIBE detects miRNA-target interactions transcriptome-wide in single cells

MicroRNAs are gene regulatory molecules that play important roles in numerous biological processes including human health. The function of a given microRNA is defined by its selection of target transcripts, yet current state-of-the-art experimental methods to identify microRNA targets are laborious and require millions of cells. We have overcome these limitations by fusing the microRNA effector protein Argonaute2 to the RNA editing domain of ADAR2, allowing for the first time the detection of microRNA targets transcriptome-wide in single cells. Our agoTRIBE method reports functional microRNA targets which are additionally supported by evolutionary sequence conservation. As a proof-of-principle, we study microRNA interactions in single cells, and find substantial differential targeting across the cell cycle. Lastly, agoTRIBE additionally provides transcriptome-wide measurements of RNA abundance and will allow the deconvolution of microRNA targeting in complex samples such as tissues at the single-cell level.

genomics↗