Understanding functional miRNA-target interactions in vivo by site-specific genome engineering.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 25135198.
- Also identified by DOI 10.1038/ncomms5640 and PMC identifier 4143950.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
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.
Medical subject headings
- Clustered Regularly Interspaced Short Palindromic Repeats
- Deoxyribonucleases
- Endonucleases
- Genetic Engineering
- MicroRNAs
- Response Elements