Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6248934

Title:Gene trap vectors
Inventors:Tessier-Lavigne, Marc; San Francisco, CA, USA
Skarnes, William C.; Berkeley, CA, USA
Mitchell, Kevin; Albany, CA, USA
Leighton, Philip A.; San Francisco, CA, USA
Summary:This invention introduces gene trap vectors comprised of a polynucleotide consisting of a promoterless selectable marker and axon reporter encoding sequences for expressing targeted gene products in vertebrate neurons. The axon reporter encoding sequences are described as being operatively joined to an internal ribosome-entry site, and may comprise a splice acceptor site located 5' to the selectable marker and axon reporter encoding sequences. Further disclosed are methods of expressing an axon reporter in a cell by transferring the vectors into an embryonic stem cell and incubating the cell under conditions whereby the cell or a progeny of the cell differentiates into a neuron comprising an axon or dendrites, such that the neuron expresses the axon reporter under the transcriptional control of the gene. Also provided are methods of detecting the axon reporter in the axon or dendrites. Further claims of the invention include neuronal specific expression which may be effected in binary systems. Therapeutic applications include the rapid screening of genes involved in establishing correct patterns of neuronal connectivity.
Abstract:The invention provides methods and compositions for expressing targeted gene products in vertebrate neurons. The compositions include gene trap vectors comprising a polynucleotide comprising promoterless selectable marker and axon reporter encoding sequences, which may be operatively joined to an internal ribosome-entry site, and may comprise a splice acceptor site located 5' to the selectable marker and axon reporter encoding sequences. The methods include methods of expressing an axon reporter in a cell by transferring the subject vectors into an embryonic stem cell and incubating the cell under conditions whereby the cell or a progeny of the cell differentiates into a neuron comprising an axon or dendrites, and the neuron expresses the axon reporter under the transcriptional control of the gene; and specifically detecting the axon reporter in the axon or dendrites. Neuronal specific expression may also be effected in disclosed binary systems.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6248934
Application Number:US1998000191652
Date Filed:13/11/1998
Date Published:19/06/2001
Assignee:The Regents of the University of California, Oakland, CA, USA


 
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