Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5728521

Title:Targeted cleavage of RNA using eukaryotic ribonuclease P and external guide sequence
Inventors:Yuan, Yan; New Haven, CT, USA
Guerrier-Takada, Cecilia; New Haven, CT, USA
Altman, Sidney; Hamden, CT, USA
Liu, Fenyong; New Haven, CT, USA
Summary:This invention describes a method of targeting ribonuclease P cleavage of RNA with an oligonucleotide comprised of a target recognition sequence and an RNAase P binding sequence. Central to the invention is the discovery that any RNA can be targeted for cleavage by RNAase P from eukaryotic cells to form a hybrid with the target RNA, thereby creating a substrate for cleavage by RNAase P in vitro. For human RNAase P, a suitably designed oligoribonucleotide is used in which the anticodon stem and loop have been deleted. Further disclosed herein are methods by which a selected RNA may be targeted for cleavage by the host cell RNAase P, thus preventing expression of the function of the target RNA. Therapeutic applications are included for treatments requiring the suppression of disease-causing genes in vivo, such as in the treatment of viral infections, bacterial infections and cancer.
Abstract:It has been discovered that any RNA can be targeted for cleavage by RNAase P from eukaryotic cells, for example, human RNAase P, using a suitably designed oligoribonucleotide (external guide sequence, or EGS) to form a hybrid with the target RNA, thereby creating a substrate for cleavage by RNAase P in vitro. The EGS hydrogen bonds to the targeted RNA to form a partial tRNA like structure including the aminoacyl acceptor stem, the T stem and loop, and part of the D stem. The most efficient EGS with human RNAase P is the EGS in which the anticodon stem and loop was deleted. Modifications can also be made within the T-loop. Methods are also disclosed to randomly select and to express a suitable EGS in vivo to make a selected RNA a target for cleavage by the host cell RNAase P, thus preventing expression of the function of the target RNA. The methods and compositions should be useful to prevent the expression of disease-causing genes in vivo.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5728521
Application Number:US1994000215082
Date Filed:18/03/1994
Date Published:17/03/1998
Assignee:Yale University, New Haven, CT, USA


 
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