Ribonuclease R (RNase R) stands as a testament to the ingenuity of genetic engineering, originating from Escherichia coli and harnessed through recombinant expression. This enzyme, pivotal in the realm of RNA research, operates as a magnesium-dependent 3' - 5' exoribonuclease, demonstrating a remarkable capability to cleave almost all linear RNA substrates. However, it intriguingly spares circular RNA, lariat RNA, and double-stranded RNA with 3' protruding ends shorter than 7 nucleotides. This unique specificity has rendered RNase R indispensable in a myriad of experimental contexts.
One of the distinguishing features of RNase R lies in its ability to efficiently eliminate linear RNA devoid of secondary structures. This characteristic has revolutionized the purification of circular RNA molecules, facilitating downstream applications such as RNA sequencing with unprecedented convenience and precision. The unit definition of RNase R, denoting the amount required to hydrolyze 1 microgram of poly-r(A) product under specified conditions, underscores its quantitative utility in experimental setups.
The applications of RNase R span a broad spectrum of RNA-focused research endeavors. Foremost among these is its role in the enrichment of circular RNA from complex biological samples. By selectively targeting linear RNA, RNase R facilitates the isolation of circular RNA species, paving the way for in-depth analyses and insights into their biological functions. Moreover, RNase R finds extensive utility in the analysis and identification of lariat intronic sequences, shedding light on the intricate mechanisms governing RNA processing and maturation.
Beyond its applications in cyclic RNA studies, RNase R has emerged as a vital tool in the exploration of alternative splicing events. By selectively degrading linear RNA transcripts, RNase R enables researchers to discern the intricate landscape of alternative splicing variants with unparalleled clarity and specificity. This capacity holds profound implications for deciphering the regulatory networks underlying gene expression and cellular differentiation.
In essence, Ribonuclease R embodies a paradigm shift in RNA research, offering unprecedented precision and versatility in the manipulation and analysis of diverse RNA species. Its remarkable specificity and efficacy have propelled it to the forefront of molecular biology, empowering researchers to unravel the complexities of RNA biology with unprecedented depth and clarity. As we delve deeper into the intricate tapestry of cellular processes, RNase R stands as a steadfast ally, illuminating the path towards new frontiers in RNA research.
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