Formulation and delivery of modified nucleoside, nucleotide, and nucleic acid compositions
The present disclosure provides, inter alia, formulation compositions comprising modified nucleic acid molecules which may encode a protein, a protein precursor, or a partially or fully processed form of the protein or a protein precursor. The formulation composition may further include a modified nucleic acid molecule and a delivery agent. The present invention further provides nucleic acids useful for encoding polypeptides capable of modulating a cell's function and/or activity.
1. A method of administering an mRNA to a subject, the method comprising:
(a) injecting the subject with the mRNA, and
(b) delivering electric pulses at or near the site of injection in three stages, wherein
(i) the first stage is a single pulse at an amplitude of approximately 450V for a pulse duration of approximately 0.05 ms and a pause interval of approximately 0.2 ms;
(ii) the second stage is a single pulse at an amplitude of approximately 450V for a pulse duration of approximately 0.05 ms and a pause interval of approximately 50 ms; and
(iii) the third stage is a pulse repeated eight times at an amplitude of approximately 11 OV for a pulse duration of approximately 10 ms and a pause interval of approximately 20 ms,
wherein the method results in an increase in the duration of protein expression at the site of injection from the mRNA in a cell or tissue of the subject compared to protein expression at the site of injection in the subject with the mRNA without electroporation.
2. The method of claim 1 , wherein the injection is selected from the group consisting of intramuscular injection and intradermal injection.
3. The method of claim 2 , wherein the injection is intramuscular injection.
4. The method of claim 1 , wherein the mRNA comprises at least one chemical modification.
5. The method of claim 4 , wherein the at least one chemical modification is selected from the group consisting of pseudouridine, 1-methylpseudouridine and 5-methylcytosine.
6. The method of claim 1 , wherein the mRNA is administered at a dose selected from the group consisting of 0.025 mg/kg, 0.25 mg/kg, 2.5 mg/kg and 5 mg/kg.