Rapid selection of accessible and cleavable sites in RNA by Escherichia coli RNase P and random external guide sequences.
A method of inhibiting the expression of particular genes by using external guide sequences (EGSs) has been improved in its rapidity and specificity. Random EGSs that have 14-nt random sequences are used in the selection procedure for an EGS that attacks the mRNA for a gene in a particular location. A mixture of the random EGSs, the particular target RNA, and RNase P is used in the diagnostic procedure, which, after completion, is analyzed in a gel with suitable control lanes. Within a few hours, the procedure is complete. The action of EGSs designed by an older method is compared with EGSs designed by the random EGS method on mRNAs from two bacterial pathogens.
Lundblad
Eirik W
EW
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Xiao
Gaoping
G
Ko
Jae-Hyeong
JH
Altman
Sidney
S
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D030561:Databases, Nucleic Acid; D004926:Escherichia coli / Q000201:enzymology*; D012333:RNA, Messenger / Q000235:genetics; D043262:Ribonuclease P / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; D012333:RNA, Messenger; D043262:Ribonuclease P