Nucleotide sequence and pharmaceutical composition based thereon with prolonged VEGF transgene expression
First Claim
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1. A method for extending the lifetime of mRNA encoding vascular endothelial growth factor (VEGF) in a host cell comprising:
- performing at least one modification selected from the group consisting of at least one pointwise single deletion of guanine, adenosine, thiamine, or cytosine, and at least one pointwise single substitution of guanine or adenosine with cytosine, in the 3′
-untranslated region of a VEGF gene to produce at least one engineered polynucleotide;
transforming the at least one engineered polynucleotide into a host cell,determining lifetime of said at least one mutant mRNA in the host cell, wherein when the engineered polynucleotide is transformed into and transcribed by the host cell, the at least one mutant mRNA is its transcription product, andselecting at least one mutant mRNA that has an extended lifetime compared to lifetime of an otherwise identical control mRNA which is a transcription product of the VEGF gene that has not been mutated.
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Abstract
A nucleic acid or transgene comprising a modified VEGF 3′-untranslated region (3′-UTR) polynucleotide sequence and a polynucleotide sequence encoding a Vascular Endothelial Growth Factor (VEGF). When transformed into a host cell, the nucleic acid or transgene exhibits a high stability and provides prolonged and reliable expression of VEGF. A method for extending the lifetime of transgene mRNA encoding VEGF in a mammalian host cell. A method for treating a subject in need of increased or modified expression of VEGF using this nucleic acid or transgene.
11 Citations
9 Claims
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1. A method for extending the lifetime of mRNA encoding vascular endothelial growth factor (VEGF) in a host cell comprising:
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performing at least one modification selected from the group consisting of at least one pointwise single deletion of guanine, adenosine, thiamine, or cytosine, and at least one pointwise single substitution of guanine or adenosine with cytosine, in the 3′
-untranslated region of a VEGF gene to produce at least one engineered polynucleotide;transforming the at least one engineered polynucleotide into a host cell, determining lifetime of said at least one mutant mRNA in the host cell, wherein when the engineered polynucleotide is transformed into and transcribed by the host cell, the at least one mutant mRNA is its transcription product, and selecting at least one mutant mRNA that has an extended lifetime compared to lifetime of an otherwise identical control mRNA which is a transcription product of the VEGF gene that has not been mutated. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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Specification