Journal
CELL REPORTS
Volume 25, Issue 4, Pages 1097-+Publisher
CELL PRESS
DOI: 10.1016/j.celrep.2018.09.084
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Funding
- IMMAGINA BioTechnology s.r.l.
- Provincia Autonoma di Trento, Italy (AxonomiX research project)
- Wellcome Trust [106098/Z/14/Z]
- SMA Trust (UK SMA Research Consortium)
- Wellcome Trust [106098/Z/14/Z] Funding Source: Wellcome Trust
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Ribosome profiling, or Ribo-seq, is based on large-scale sequencing of RNA fragments protected from nuclease digestion by ribosomes. Thanks to its unique ability to provide positional information about ribosomes flowing along transcripts, this method can be used to shed light on mechanistic aspects of translation. However, current Ribo-seq approaches lack the ability to distinguish between fragments protected by either ribosomes in active translation or inactive ribosomes. To overcome this possible limitation, we developed RiboLace, a method based on an original puromycin-containing molecule capable of isolating active ribosomes by means of an antibody-free and tag-free pull-down approach. RiboLace is fast, works reliably with low amounts of input material, and can be easily and rapidly applied both in vitro and in vivo, thereby generating a global snapshot of active ribosome footprints at single nucleotide resolution.
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