Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6319707

Title:Cap-independent multicistronic retroviral vectors
Inventors:Adam, Mohammed A.; Kirkland, Canada
Summary:Described herein are retroviral virions for producing coordinately expressed polycistronic mRNA in transfected host cells. The invention relates to a representative retroviral construct capable of forming a proviral genome in a host cell containing a first nucleotide coding sequence, a second nucleotide coding sequence, and a third nucleotide sequence capable of hybridizing under stringent conditions to a 5' nontranslated region (NTR) of a picornavirus RNA or its complementary RNA strand, such that transcription of the proviral genome gives rise to a messenger RNA molecule containing transcripts of the first, second, and third nucleotide sequences. Further claims of the invention include the transcript of the third nucleotide sequence in the messenger RNA molecule which is described as containing a nucleic acid capable of forming a regulatory stem-loop nucleic acid structure followed by at least one operable AUG start codon. Further disclosed are properties by which the regulatory stem-loop nucleic acid structure is capable of operably binding to a translation initiation complex in a host cell such that the transcripts of the first and second nucleotide sequences in the messenger RNA molecule are coordinately expressed in the host cell.
Abstract:Retroviral vectors for producing coordinately expressed polycistronic mRNA in transfected host cells. A representative retroviral construct capable of forming a proviral genome in a host cell contains a first nucleotide coding sequence, a second nucleotide coding sequence, and a third nucleotide sequence capable of hybridizing under stringent conditions to a 5' nontranslated region (NTR) of a picornavirus RNA or its complementary RNA strand. The first, second, and third nucleotide sequences are operably linked such that transcription of the proviral genome gives rise to a messenger RNA molecule containing transcripts of the first, second, and third nucleotide sequences. The transcript of the third nucleotide sequence in the messenger RNA molecule contains a nucleic acid capable of forming a regulatory stem-loop nucleic acid structure followed by at least one operable AUG start codon. The regulatory stem-loop nucleic acid structure is capable of operably binding a translation initiation complex in a host cell such that the transcripts of the first and second nucleotide sequences in the messenger RNA molecule are coordinately expressed in the host cell.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6319707
Application Number:US1993000009338
Date Filed:26/01/1993
Date Published:20/11/2001
Assignee:Fred Hutchinson Cancer Research Center Board Regents of the University of Washington, Seattle, WA, USA


 
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