RNAs play many essential roles in gene expression and are involved in various human diseases. Although genome editing technologies have been established, the engineering of sequence-specific RNA-binding proteins that manipulate particular cellular RNA molecules is immature, in contrast to nucleotide-based RNA manipulation technology, such as siRNA- and RNA-targeting CRISPR/Cas. Here, we demonstrate a versatile RNA manipulation technology using pentatricopeptide-repeat (PPR)-motif-containing proteins. First, we developed a rapid construction and evaluation method for PPR-based designer sequence-specific RNA-binding proteins. This system has enabled the steady construction of dozens of functional designer PPR proteins targeting long 18 nt RNA, which targets a single specific RNA in the mammalian transcriptome. Furthermore, the cellular functionality of the designer PPR proteins was first demonstrated by the control of alternative splicing of either a reporter gene or an endogenous CHK1 mRNA. Our results present a versatile protein-based RNA manipulation technology using PPR proteins that facilitates the understanding of unknown RNA functions and the creation of gene circuits and has potential for use in future therapeutics.
We developed a concrete floor finishing robot (the SURF ROBO) in order to eliminate heavy-duty work and increase the efficiency of concrete floor finishing work. For about one year after development, several SURF ROBOs have been used at the practical construction sites and satisfactory results were obtained in respect of quality and efficiency.Two (2 ) sets of four ( 4) piece trowels are provided , the trowels rotate reversely against each other around the caterpillars and the trowels make the concrete floor smooth as well fine as by the skilled worker.After practicing with this robot, we found that it has the capacity for finishing of a concrete -floor surface of 300 m2 per hour and it provided the same effect of a concrete-floor direct finish with precision as a plasterer . But, as working time can not be reduced by the SURF ROBO because of bleeding water , we have been developing the dehydration robot.
We analyzed modes of action of ribonuclease P (RNase P) proteins, C5 in Escherichia coli and Rpr2 in Saccharomyces cerevisiae, using a pair of complementary fluorescence-labeled oligoribonucleotides. Fluorescence resonance energy transfer-based assays revealed that RNA annealing and strand displacement activities found in archaeal RNase P proteins are prevalent in eubacterial (C5) and eukaryotic (Rpr2) RNase P proteins.
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