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Understanding funct...
Understanding functional miRNA-target interactions in vivo by site-specific genome engineering
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Bassett, Andrew R. (author)
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Azzam, Ghows (author)
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Wheatley, Lucy (author)
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Tibbit, Charlotte (author)
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Rajakumar, Timothy (author)
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McGowan, Simon (author)
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Stanger, Nathan (author)
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- Ewels, Philip Andrew (author)
- Stockholms universitet,Institutionen för biokemi och biofysik
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Taylor, Stephen (author)
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Ponting, Chris P. (author)
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Liu, Ji-Long (author)
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Sauka-Spengler, Tatjana (author)
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Fulga, Tudor A. (author)
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(creator_code:org_t)
- 2014-08-19
- 2014
- English.
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In: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 5, s. 4640-
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Abstract
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- 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.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Bassett, Andrew ...
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Azzam, Ghows
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Wheatley, Lucy
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Tibbit, Charlott ...
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Rajakumar, Timot ...
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McGowan, Simon
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show more...
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Stanger, Nathan
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Ewels, Philip An ...
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Taylor, Stephen
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Ponting, Chris P ...
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Liu, Ji-Long
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Sauka-Spengler, ...
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Fulga, Tudor A.
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show less...
- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
- Articles in the publication
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Nature Communica ...
- By the university
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Stockholm University