Court directed RNA using ribonuclease p eukaryotic and external guide sequence.

摘要

IT HAS BEEN DISCOVERED THAT ANY RNA MAY BE TERMINATED BY SPLITTING RNAase PA FROM eukaryotic cells, FOR EXAMPLE, Hela HUMANAS using a oligoribonucleotide PROPERLY DESIGNED ( "external guide sequence, or EGS) to form a hybrid with RNA DESTINATION , SO ESTABLISHING A substrate for CLEAVAGE BY RNase P in vitro. Two classes of EGS that can lead RNA BY SPLITTING FOR HUMAN RNase. These are prepared by transcription of a small fragment of DNA containing a sequence encoding an RNA which can assume a secondary structure TYPE tRNA. IN THE FIRST CLASS, STRUCTURE lacks at least the first thirteen nucleotides from the 5 '' of type tRNA SEQUENCE, STEMS AND BOWS anticodon, LOOP VARIABLE OR PART OF THE LOOP VARIABLE . The most efficient EGS RNase P WITH HUMAN IS THE EGS IN WHICH THE STALK AND HAVE BEEN DELETED anticodon loop. In the second class, the EGS HAVE CHANGES IN BOTH THE EQUIVAL TY LOOP ENTE STALK SEGMENT TYPE anticodon tRNA EGS. ALSO DESCRIBED METHODS FOR RANDOMLY SELECT AND PROPER express EGS, LIVE TO MAKE A SELECTED BY OBTAINING RNA unfolding of RNase P host cell, thus preventing expression of the function of RNA TARGET. The methods and compositions should be used to prevent expression of genes that cause illness LIVE.