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In plants, microRNA (miRNA) target MIMICs (MIMs) have been widely used to inhibit miRNA function. They are based on the Arabidopsis INSENSITIVE TO PHOSPATE STARVATION 1 (IPS1) gene that corresponds to a non-coding RNA containing a miR399 binding site that can be modified to sequester and inhibit any miRNA of interest. However, the efficacy of miRNA inhibition of these different MIMs can vary greatly. Using MIMs that have strong efficacy (MIM159) and poor efficacy (MIM165), we investigate the underlying cause of this variation. Firstly, sequence alignments of IPS1 homologs from the Brassicaceae identified a highly conserved sequence immediately downstream of the miRNA binding site. Mutating this sequence in the context of the MIM159 attenuates its strong efficacy. This conserved flanking region contains a predicted stem-loop structure that is also predicted to be present in most modified MIMs that appear to have a strong efficacy, but not in MIM165 that has a poor efficacy. Restoring this predicted stem-loop in MIM165 via mutation of only three or five nucleotides within the conserved flanking region resulted in MIM165 variants that have very strong efficacies of miRNA inhibition. However, specifically mutating this predicted stem-loop in the MIM159 context failed to significantly reduce efficacy, and additional mutations to restore this predicted stem-loop weakened efficacy further. Although this shows there is no simple correlation between this predicted stem-loop and efficacy, these results add to the growing evidence that the sequence context of miRNA binding sites is important, and that minor nucleotide substitutions to flanking sequences of miRNA binding sites can strongly enhance or attenuate the miRNA-target interaction.
Pubmed ID: 31245688
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Harvester is a Web-based tool that bulk-collects bioinformatic data on human proteins from various databases and prediction servers. It is a meta search engine for gene and protein information. It searches 16 major databases and prediction servers and combines the results on pregenerated HTML pages. In this way Harvester can provide comprehensive gene-protein information from different servers in a convenient and fast manner. As full text meta search engine, similar to Google trade mark, Harvester allows screening of the whole genome proteome for current protein functions and predictions in a few seconds. With Harvester it is now possible to compare and check the quality of different database entries and prediction algorithms on a single page. Sponsors: This work has been supported by the BMBF with grants 01GR0101 and 01KW0013.
View all literature mentionsIt is used to compare a novel sequence with those contained in nucleotide and protein databases by aligning the novel sequence with previously characterized genes.
View all literature mentionsA commercial organization which provides assay technologies to isolate DNA, RNA, and proteins from any biological sample. Assay technologies are then used to make specific target biomolecules, such as the DNA of a specific virus, visible for subsequent analysis.
View all literature mentionsSoftware package as multiple alignment program for amino acid or nucleotide sequences. Can align up to 500 sequences or maximum file size of 1 MB. First version of MAFFT used algorithm based on progressive alignment, in which sequences were clustered with help of Fast Fourier Transform. Subsequent versions have added other algorithms and modes of operation, including options for faster alignment of large numbers of sequences, higher accuracy alignments, alignment of non-coding RNA sequences, and addition of new sequences to existing alignments.
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