Nuclease resistant polynucleotides and uses thereof

Abstract

The invention provides, among other things, methods of mRNA stabilizing mRNA and nuclease resistant mRNA prepared in accordance with such methods. In certain embodiments, the nuclease resistant mRNA encodes a functional protein, such as enzyme, and is characterized by its resistance to nuclease digestion, increased half-life and/or its ability to produce increased amounts of the functional protein (e.g., enzyme) encoded thereby.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Provisional Application No. 61 / 657,465, filed on Jun. 8, 2012, the disclosure of which is incorporated herein by reference.BACKGROUND[0002]The administration of exogenous nucleic acids and polynucleotides, for example DNA vectors and plasmids, to a subject for the treatment of protein or enzyme deficiencies represents a significant advance in the treatment of such deficiencies however, the administration of such exogenous nucleic acids to a subject remains especially challenging. For example, gene therapies that rely on viruses to carry and deliver exogenous polynucleotides (e.g., DNA) to host cells and that cause the integration of such polynucleotides into the host cells' genome are capable of eliciting serious immunological and inflammatory responses. Furthermore, in certain instances the integration of such exogenous polynucleotides into the host cells' genome has the potential of misregulating the expression ...

AI Technical Summary

Benefits of technology

[0021]Also provided herein are methods of increasing the translational efficiency of an exogenous mRNA transcript. Such methods may facilitate, for example, an increase in the production of an expression product produced following translation of the mRNA polynucleotides or transcripts of the present inventions. Generally, such methods comprise a step of contacting the mRNA polynucleotide transcript with a stabilizing oligonucleotide that is complementary to the coding and/or non-coding region of the mRNA transcript under suitable conditions (e.g., high stringency conditions), thereby causing the mRNA p

Problems solved by technology

The administration of exogenous nucleic acids and polynucleotides, for example DNA vectors and plasmids, to a subject for the treatment of protein or enzyme deficiencies represents a significant advance in the treatment of such deficiencies however, the administration of such exogenous nucleic acids to a subject remains especially challenging.

Furthermore, in certain instances the integration of such exogenous polynucleotides into the host cells' genome has the potential of misregulating the expression of the host's endogenous genes and unpredictably impacting cellular activity.

Such plasmids are however, frequently characterized as having highly inefficient cellular uptake in vivo.

While in some instances, the use of recombinant proteins and enzymes may provide a means of ameliorating the symptoms of the underlying deficiency, the utility of such therapies are often limited and are not considered curative.

Furthermore, recombinant proteins or enzymes are often prepared using non-human cell lines and may lack certain post-translational modifications (e.g., human glycosylation) relative to their endogenously produced counterparts.

Recombinant protein and enzyme replacement therapies are also associated with great financial expense.

Since replacement therapie

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