Engineered nucleic acids and methods of use thereof

Abstract

Provided are compositions and methods for delivering biological moieties such as modified nucleic acids into cells to modulate protein expression. Such compositions and methods include the use of modified messenger RNAs, and are useful for production of proteins.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]The present application claims the benefit of U.S. Provisional Application No. 61 / 404,413, filed Oct. 1, 2010, the disclosure of which is considered part of (and is incorporated herein by reference in) the disclosure of this application.BACKGROUND OF THE INVENTION[0002]Naturally occurring RNAs are synthesized from four basic ribonucleotides: ATP, CTP, UTP and GTP, but may contain post-transcriptionally modified nucleotides. Further, approximately one hundred different nucleoside modifications have been identified in RNA (Rozenski, J, Crain, P, and McCloskey, J. (1999). The RNA Modification Database: 1999 update. Nucl Acids Res 27: 196-197). The role of nucleoside modifications on the immuno-stimulatory potential and on the translation efficiency of RNA, however, is unclear.[0003]There are multiple problems with prior methodologies of effecting protein expression. For example, heterologous DNA introduced into a cell can be inherited by dau...

AI Technical Summary

Benefits of technology

[0009]In one aspect, the disclosure features a method of increasing the production of a recombinantly expressed protein of interest in a cell, comprising the steps: (i) providing a target cell comprising a recombinant nucleic acid encoding the protein of interest; and (ii) introducing into the target cell a composition comprising a first isolated nucleic acid comprising a translatable region encoding a translation effector protein and a nucleoside modification under conditions such that the effector protein is produced in the cell, thereby increasing the production of the recombinantly expressed protein in the cell.

[0010]In some embodiments, the target cell is a mammalian cell. In some embodiments, the target cell is a yeast cell. In some embodiments, the target cell is a bacterial cell, an insect cell, or a plant cell. In some embodiments, the protein of interest is a secreted protein. In some embodiments, the protein of interest is a transmembrane protein. In some embodiments, the protein of interest is an antibody or an antigen-binding fragment thereof. In some embodiments, the protein of interest is a growth factor or

Problems solved by technology

There are multiple problems with prior methodologies of effecting protein expression.

Introduced DNA can integrate into host cell genomic DNA at some frequency, resulting in alterations and/or damage to the host cell genomic DNA.

Further, it is difficult

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