Hairpin MRNA elements and methods for the regulation of protein translation
AssigneeUniversity of Florida Research Foundation, Inc.
InventorsChen Ling, Arun Srivastava
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Abstract
The present invention provides novel mRNA elements capable of forming hairpin, double-stranded RNA structures independent of other non-coding RNAs. These mRNAs are stably expressed, lack polyadenylation tails, and allow minimal protein translation except when in the presence of specific proteins. Also provided, are compositions and kits comprising the mRNA element, as well as methods for its use in the regulation of protein translation. Advantageously, the disclosed elements represent a novel tool useful in regulating the expression of a wide variety of proteins of interest.
Claims22
A method for providing a mammal with a therapeutically-effective amount of a biologically-active mammalian polypeptide to a mammal in need thereof, comprising introducing into suitable cells of a mammal in need thereof, an effective amount of a recombinant adeno-associated viral (rAAV) vector, for a time sufficient to produce a biologically-active mammalian polypeptide therefrom in a population of cells or a tissue of the mammal into which the rAAV vector has been introduced, wherein the rAAV vector comprises an expression cassette that comprises a promoter operably linked to a nucleic acid segment that encodes the biologically-active mammalian polypeptide, wherein the expression cassette lacks a polyA signal sequence, and wherein the mRNA produced from the vector lacks a polyA sequence at its 3′-end but includes an inverted terminal repeat (ITR) at its 3′-end.
A method for increasing the level of a biologically-active protein or polypeptide in one or more cells of a mammal, comprising introducing a recombinant adeno-associated viral (rAAV) vector into a population of cells of the mammal, in an amount and for a time effective to increase the level of a biologically-active protein or polypeptide in one or more cells of the mammal, wherein the rAAV vector comprises an expression cassette that comprises a promoter operably linked to a nucleic acid segment that encodes the biologically-active protein or polypeptide, wherein the expression cassette lacks a polyA signal sequence, and wherein the mRNA produced from the vector lacks a polyA sequence at its 3′-end but includes an inverted terminal repeat (ITR) at its 3′-end.
A method for expressing a selected polypeptide in a cell, comprising introducing into the cell a population of rAAV virions comprising a recombinant adeno-associated viral (rAAV) vector, and expressing the vector in the cell, wherein the rAAV vector comprises an expression cassette that comprises a promoter operably linked to a nucleic acid segment that encodes the biologically-active protein or polypeptide, wherein the expression cassette lacks a polyA signal sequence, and wherein the mRNA produced from the vector lacks a polyA sequence at its 3′-end but includes an inverted terminal repeat (ITR) at its 3′-end.
Dependencies are inferred from the claim text.