During the cross-referencing process, we observed numerous examples of inconsistent genome annotation in different S. These novel sRNAs were annotated, cross-referenced in the genomes of strains NCTC 8325 and MRSA252, and added to the newly created genome files as well. Examining the data, we identified 39 novel putative sRNAs that had not been previously reported. To demonstrate the application of these new files, we performed RNA-seq on CA-MRSA strain USA300 growing in laboratory media and human serum and aligned the data to our newly created sRNA annotated genome files. aureus research community in the form of our newly generated annotation files, while at the same time presenting the first examination of differential sRNA expression in pathophysiologically relevant conditions. This study provides an invaluable resource to the S. aureus and demonstrate that the newly identified tsr25 is the most highly upregulated sRNA in human serum. Finally, using our newly created annotation files as a reference, we perform a global analysis of sRNA gene expression in S. aureus, which may be hindering the analysis and functional exploration of these elements. Investigating the genomic loci of the newly identified sRNAs revealed a surprising degree of inconsistency in genome annotation in S. This analysis led to the discovery of 39 previously unidentified sRNAs. aureus, were then used as a foundation to identify novel sRNAs in the community-associated methicillin-resistant strain USA300. These files, the first to include genome-wide annotation of sRNAs in S. aureus strains (MRSA252, NCTC 8325, and USA300), in which we added annotations for >260 previously identified sRNAs. aureus, we generated updated GenBank files for three commonly used S. To facilitate improved detection and analysis of sRNAs in S. sRNA genes are typically absent from genome annotation files, and as a consequence, their existence is often overlooked, particularly in global transcriptomic studies. In Staphylococcus aureus, hundreds of small regulatory or small RNAs (sRNAs) have been identified, yet this class of molecule remains poorly understood and severely understudied.
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