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RNA probes are usually prepared by in vitro transcription (see Fig. 1 above). The RNA probe is transcribed from a linear DNA template using highly specific bacteriophage DNA-dependant RNA polymerases from the Salmonella bacteriophage SP6, and the E. coli. bacteriophages T3 and T7 (RNA polymerase T7, T3 or SP6).
Activated fluorescent RNA probes, their expression vectors, and detection methods
PendingCN122081459Asimple wayeasy to operateMicrobiological testing/measurementDNA/RNA fragmentationIntracellularLiving cell
This application discloses a technique for detecting target nucleic acid molecules using an activated fluorescent RNA probe, its expression vector, and a detection method. It includes: probe design and preparation methods; labeling and detection of target nucleic acid molecules in solution samples using the probe; labeling and detection of target nucleic acid molecules in living cells using the probe; and labeling and detection of target nucleic acid molecules in immobilized cells using the probe. This invention overcomes the shortcomings of current commercially synthesized FISH and molecular beacon probes, such as their inability to be genetically encoded, difficult preparation, and high cost. It provides an efficient, simple, and rapid method to obtain activated fluorescent RNA probes for detecting target nucleic acid molecules through genetic encoding or in vitro transcription. This offers a practical tool for research on fundamental life science issues and for point-of-care and even real-time diagnosis of diseases, possessing broad scientific and social significance.
Owner:EAST CHINA UNIV OF SCI & TECH