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The participation of tRNAs in fundamental aspects of biology and disease necessitates an accurate, experimentally confirmed annotation of tRNA genes and curation of tRNA sequences. This has been challenging because RNA secondary structure, nucleotide modifications, and tRNA gene multiplicity complicate sequencing and mapping efforts. To address these issues, we developed hydro-tRNAseq, a method based on partial alkaline RNA hydrolysis that generates fragments amenable for sequencing. To identify transcribed tRNA genes, we further complemented this approach with photoactivatable crosslinking and immunoprecipitation (PAR-CLIP) of SSB/La, a conserved protein involved in pre-tRNA processing. Our results show that approximately half of all predicted tRNA genes are transcribed in human cells. We also report nucleotide modification sites and their order of introduction, and we identify tRNA leaders, trailers, and introns. By using complementary sequencing-based methodologies, we present a human tRNA atlas and determine expression levels of mature and processing intermediates of tRNAs in human cells.




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Last April, Senior U.S. District Judge Stephen Limbaugh ruled that computer and video games had "no conveyance of ideas, expression or anything else that could possibly amount to free speech" in a St. Louis County case that sought to limit children's access to mature video games.


Although he provides some examples that hint at the power of this new mode of expression, examples pulled from popular culture (MTV clips, Oliver Stone's Natural Born Killers, the first 96 seconds of an ABC News documentary), he contends that the movement is just now starting to unfold.


Once mature, video will be able to communicate highly complex ideas and thought without words; it will be "a form of communication powerful enough to help us fashion new understandings, stronger understandings."


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