Description du projet
Une nouvelle technologie pour le séquençage des ARN non-codants
Le séquençage de l’ARN est une technique qui permet l’analyse quantitative des molécules d’ARN messager dans un échantillon biologique. Toutefois, il est limité aux ARN codant les protéines et ne peut pas fournir la séquence d’autres molécules d’ARN comme les petits ARN non-codants. Le projet droplet-small-seq, financé par l’UE, entend aborder cette limitation et offrir un aperçu de l’expression et de la fonction des microARN dans les cellules individuelles. L’équipe développera une technologie basée sur les gouttelettes à haut débit pour capturer et séquencer les microARN et les ARNm en même temps. Étant donné le rôle régulatoire des ARN non-codants dans l’expression génétique, l’approche du projet devrait identifier de nombreuses applications utiles dans la recherche.
Objectif
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.
Champ scientifique
Mots‑clés
Programme(s)
Régime de financement
MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinateur
01513 Vilnius
Lituanie