Method for identification of polynucleotides capable of cleaving target mRNA sequences

• Main Authors: SULLIVAN JOHN M, YAU EDWIN H
• Format: Patent
• Language: English
• Published: 18.09.2008
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

Provided are methods for identifying a polynucleotide that can reduce the level of a target mRNA. The method can be performed by providing cells that express an RNA polynucleotide that contains a target mRNA sequence, an internal ribosome entry sequence (IRES) and a sequence encoding a secreted reporter protein; introducing to the cells a test polynucleotide, and measuring activity of the secreted reporter protein. A reduction in secreted reporter protein activity relative to a control cell into which the test polynucleotide has not been introduced is indicative that the test polynucleotide is capable of reducing the level of the target mRNA in the cells. The method is adaptable for high throughput screening methods and is suited for identifying polynucleotides that can catalyze cleavage of target mRNA and/or act on target mRNA through and antisense or RNAi mechanism.