Descripción del proyecto
Nueva tecnología para secuenciar ARN no codificantes
La secuenciación del ARN es una técnica que posibilita el análisis cuantitativo de las moléculas de ARN mensajero en una muestra biológica. Sin embargo, se limita a los ARN codificantes de proteínas y no puede proporcionar la secuencia de otras moléculas de ARN, como los ARN no codificantes pequeños. El proyecto droplet-small-seq, financiado con fondos europeos, tiene como objetivo abordar esta limitación y ofrecer datos sobre la expresión y la función de los microARN en células individuales. El equipo desarrollará una tecnología de alto rendimiento basada en gotas para capturar y secuenciar microARN y ARNm al mismo tiempo. Dado el papel regulador de los ARN no codificantes en la expresión génica, se espera que el método del proyecto halle muchas aplicaciones útiles en la investigación.
Objetivo
Droplet-based single-cell RNA-sequencing (scRNA-seq) technologies have penetrated almost all branches of life sciences and have significantly advanced our understanding of cellular processes and organism development. However, despite their astonishing impact, most of the scRNA-seq technologies reported to date rely on poly(A) tail capture and thus are mainly restricted to the protein-coding RNAs, while neglecting a substantial proportion of the transcriptome, including small non-coding RNAs. As a result, very little is known about the non-coding RNA expression and function in individual cells, and especially their role in the establishment of cellular phenotypic diversity. Small RNAs contain a variety of classes, of which miRNAs are the most common and these act as regulatory molecules by suppressing translation of mRNAs. In addition, loss-of-function studies of miRNAs uncovered their involvement in development of nearly all tissues, including hematopoiesis. However, most studies exploring miRNA dynamics reported to date relied on bulk cell assays, thus disregarding the individual cell types and their heterogeneity. In the scope of this proposal, we aim to develop a high-throughput droplet-based single-cell small RNA-seq (droplet-small-seq) for simultaneous miRNA and mRNA capture and sequencing. We will apply this newly developed technique to investigate the regulatory roles of miRNAs in cell fate decision during hematopoietic development at a single-cell level.
Ámbito científico
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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinador
01513 Vilnius
Lituania