Liver Specific Delivery of Messenger RNA
Disclosed herein are compositions and methods of modulating the expression of gene or the production of a protein by transfecting target cells with nucleic acids. The compositions disclosed herein demonstrate a high transfection efficacy and are capable of ameliorating diseases associated with protein or enzyme deficiencies.
1 - 126 . (canceled)
127 . A method of delivery of messenger RNA (mRNA) for in vivo production of protein in the liver, comprising administering systemically to a subject in need thereof a composition comprising a first mRNA encoding a protein, encapsulated within a liposome such that the administering of the composition results in the prolonged stable expression of the protein encoded by the mRNA in the liver, and a second nucleic acid; and wherein the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids and has a size less than about 150 nm.
128 . A composition comprising an mRNA encoding a protein, encapsulated within a liposome such that the administering of the composition results in the prolonged stable expression of the protein encoded by the mRNA in the liver; and wherein the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids and has a size less than about 150 nm.
129 . The method of claim 127 , wherein the one or more non-cationic lipids comprise DOPE.
130 . The method of claim 127 , wherein the one or more cationic lipids comprise DLinDMA, CLinDMA, or DLin-K-XTC2-DMA.
131 . The method of claim 127 , wherein the one or more PEG-modified lipids comprise DMG-PEG-2000 or C8-PEG-2000.
132 . The method of claim 127 , wherein the mRNA is of at least 30 kDa.
133 . The method of claim 127 , wherein the mRNA is modified to enhance stability.
134 . The method of claim 133 , wherein the mRNA is modified to include a modified nucleotide, an alteration to the 5′ or 3′ untranslated region, a cap structure or a poly A tail.
135 . The method of claim 134 , wherein the modified nucleotide is a pseudouridine.
136 . The method of claim 127 , wherein the mRNA is unmodified.
137 . The method of claim 127 , wherein the protein is secreted extracellularly after expression.
138 . The method of claim 127 , wherein the composition is administered intravenously.
139 . The method of claim 127 , wherein the protein is systemically distributed.
140 . The method of claim 127 , wherein the expression of the protein encoded by the mRNA in the liver is detectable at least 4 hours post administration.
141 . The method of claim 127 , wherein the composition comprises two or more mRNAs encoding two or more distinct proteins.
142 . The method of claim 127 , wherein the one or more cationic lipids carry a net positive charge at physiological pH.
143 . The method of claim 142 , wherein the one or more cationic lipids constitute about 20-70% mol % of the total lipids present in the liposome.
144 . The method of claim 127 , wherein the one or more PEG-modified lipids comprise a polyethylene glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C 6 -C 20 length.
145 . The method of claim 127 , wherein the one or more PEG-modified lipids constitute up to about 20 mol % of the total lipids present in the liposome.