Projektbeschreibung
Mit neuer Technologie nicht-kodierende RNA sequenzieren
Das Verfahren der RNA-Sequenzierung dient der quantitativen Analyse von Boten-RNA-Molekülen in einer biologischen Probe. Es ist jedoch auf die proteinkodierenden RNA-Varianten beschränkt und kann keine Sequenzen anderer RNA-Moleküle wie etwa kleiner nicht-kodierender RNA liefern. Ziel des EU-finanzierten Projekts droplet-small-seq ist, diese Einschränkung zu beseitigen sowie Einblicke in die Expression und Funktion von microRNA in einzelnen Zellen zuzulassen. Das Team wird eine auf Tröpfchen basierende Hochdurchsatztechnologie zur gleichzeitigen Erfassung und Sequenzierung von microRNA und mRNA entwickeln. Angesichts der regulatorischen Funktion der nicht-kodierenden RNA bei der Genexpression ist zu erwarten, dass es für die Herangehensweise des Projekts vielfach nützliche Anwendungsmöglichkeiten in der Forschung geben wird.
Ziel
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.
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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Koordinator
01513 Vilnius
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