Understanding functional miRNA-target interactions in vivo by site-specific genome engineering
2014 (English)In: Nature Communications, ISSN 2041-1723, Vol. 5, 4640- p.Article in journal (Refereed) Published
MicroRNA (miRNA) target recognition is largely dictated by short 'seed' sequences, and single miRNAs therefore have the potential to regulate a large number of genes. Understanding the contribution of specific miRNA-target interactions to the regulation of biological processes in vivo remains challenging. Here we use transcription activator-like effector nuclease (TALEN) and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 technologies to interrogate the functional relevance of predicted miRNA response elements (MREs) to post-transcriptional silencing in zebrafish and Drosophila. We also demonstrate an effective strategy that uses CRISPR-mediated homology-directed repair with short oligonucleotide donors for the assessment of MRE activity in human cells. These methods facilitate analysis of the direct phenotypic consequences resulting from blocking specific miRNA-MRE interactions at any point during development.
Place, publisher, year, edition, pages
2014. Vol. 5, 4640- p.
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:su:diva-107811DOI: 10.1038/ncomms5640ISI: 000341057000031OAI: oai:DiVA.org:su-107811DiVA: diva2:752589