Aldehyde dehydrogenase 2 enzymes, alcohol dehydrogenase 1 enzymes, nucleic acids encoding enzymes, lipid nanoparticles, and methods of use

Abstract

Alcohol dehydrogenase 1 enzymes, aldehyde dehydrogenase 2 enzymes, nucleic acids encoding alcohol dehydrogenase 1 enzymes and aldehyde dehydrogenase 2 enzymes, lipid nanoparticles, lipid nanoparticles comprising nucleic acids encapsulated therein, pharmaceutical compositions comprising lipid nanoparticles which comprise nucleic acids encapsulated therein are useful for treating alcohol-related conditions, such as alcohol poisoning.

Description

PATENT Docket No.: 048440-877001WO ALDEHYDE DEHYDROGENASE 2 ENZYMES, ALCOHOL DEHYDROGENASE 1 ENZYMES, NUCLEIC ACIDS ENCODING ENZYMES, LIPID NANOPARTICLES, AND METHODS OF USE CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to US Application No.63 / 664,296 filed June 26, 2024, the disclosure of which is incorporated by reference herein in its entirety. REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAM LISTING APPENDIX SUBMITTED AS AN ASCII FILE

[0002] A Sequence Listing in an XML file having the title “048440-877001WO-Sequence- Listing-ST26,” having 1,050,227 bytes and created on June 19, 2025, is incorporated by reference herein in its entirety. BACKGROUND

[0003] Excessive alcohol consumption, such as binge drinking, can lead to alcohol poisoning and death. It is estimated that more than 140,000 people die from alcohol-related causes annually, making alcohol the fourth-leading preventable cause of death in the United States behind tobacco, poor diet and physical inactivity, and illegal drugs. In 2014, there were just under 5 million alcohol-related ER visits in the US. There were 61 percent more alcohol-related emergency room visits in 2014 than in 2006. Excessive alcohol consumption and poisoning is a serious health problem not just in the US but globally. There is a need in the art for an antidote for alcohol poisoning. The disclosure is directed to this, as well as other, important ends. BRIEF SUMMARY

[0004] Provided herein are nucleic acids encoding an aldehyde dehydrogenase 2 enzyme; nucleic acids encoding alcohol dehydrogenase 1 enzyme; and nucleic acids encoding an aldehyde dehydrogenase 2 enzyme and an alcohol dehydrogenase 1 enzyme. Provided herein are aldehyde dehydrogenase 2 enzymes and alcohol dehydrogenase 1 enzymes.

[0005] Provided herein are lipid nanoparticles comprising a nucleic acid encoding an aldehyde dehydrogenase 2 enzyme; lipid nanoparticles comprising a nucleic acid encoding an alcohol dehydrogenase 1 enzyme; lipid nanoparticles comprising a nucleic acid encoding an aldehyde dehydrogenase 2 enzyme and an alcohol dehydrogenase 1 enzyme; and lipid nanoparticles comprising a first nucleic acid encoding an aldehyde dehydrogenase 2 enzyme and a second nucleic acid encoding an alcohol dehydrogenase 1 enzyme.

[0006] Provided herein are a plurality of lipid nanoparticles comprising: (i) a first lipid nanoparticle comprising a first nucleic acid encoding an aldehyde dehydrogenase 2 enzyme; and(ii) a second lipid nanoparticle comprising a second nucleic acid encoding an alcohol dehydrogenase 1 enzyme.

[0007] Provided herein are methods of treating or preventing alcohol poisoning, treating or preventing a symptom of alcohol poisoning, and treating or preventing alcohol flush syndrome in a patient in need thereof comprising administering to the patient an effective amount of the nucleic acids described herein, the enzymes described herein, the lipid nanoparticle nanoparticles described herein, and the pharmaceutical compositions described herein.

[0008] These and other embodiments of the disclosure are described herein. DESCRIPTION OF THE DRAWINGS

[0009] FIGS.1A-1B show the results of an alcohol behavior study in mice administered: (i) a combination of ALDH2*1 and ADH1B*2 (2:1) mRNA-SM102-LNP, (ii) ALDH2*1 mRNA- SM102-LNP, or (iii) PBS.

[0010] FIGS.2A-2C are graphs showing the behavioral score of mice injected with control LNP or LNP- ADH1B*2 mRNA.

[0011] FIGS.3A-3C are graphs showing the behavioral score of mice injected with control LNP or LNP- ALDH2*1 mRNA.

[0012] FIGS.4A-4C are graphs showing the behavioral score of mice injected with control LNP or LNP- ALDH2*1 mRNA.

[0013] FIGS.5A-5C are graphs showing the behavioral score of mice injected with control LNP or LNP- ALDH2*1 mRNA.

[0014] FIGS.6A-6C are graphs showing the behavioral score of mice injected with control LNP or LNP- ALDH2*1 mRNA.

[0015] FIGS.7A-7C are graphs showing plasma AST and plasma ALT levels after injection with PBS, control LNP, LNP-GFP (green fluorescent protein), or LNP-ALDH2*1 mRNA.

[0016] FIGS.8A-8B are graphs showing the behavioral score of mice injected with PBS, control LNP, or LNP-ADH1B*2 mRNA.

[0017] FIGS.9A-9B are graphs showing the body temperature of mice injected with PBS, control LNP, or LNP-ADH1B*2 mRNA.

[0018] FIG.10 is a graph showing the OD 450 for 20-120 minutes for LNP-ALDH2*1, LNP- GFP, no LNP, and blank.

[0019] FIG.11 is a graph showing the ADH activity of ADH1B*2 mRNA and GFP mRNA at OD 565 for 30 minutes.

[0020] FIG.12 is a graph showing the theoretical results of successful expression of functional ADH1B*2 and ALDH2*1 in hepatocytes via mRNA-LNP. Ethanol clearance canincrease 3-10x over baseline. DETAILED DESCRIPTION

[0021] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. See, e.g., Singleton et al., Dictionary of Microbiology and Molecular Biology, 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of this disclosure. The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0022] “Aldehyde dehydrogenase 2 enzyme” or “ALDH2 enzyme” or “Aldh2 enzyme” includes any of the recombinant or naturally-occurring forms of the ALDH2 enzyme or variants or homologs thereof that maintain ALDH2 enzyme activity (e.g., within at least 80%, at least 90%, at least 95%, or 100% activity compared to the ALDH2 enzyme. In aspects, the variants or homologs have at least 90%, 95%, or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150, or 200 continuous amino acid portion) compared to the naturally occurring ALDH2 enzyme. In embodiments, the ALDH2 enzyme is identified by the UniProt reference P05091, a homolog, or a functional fragment thereof. In embodiments, the ALDH2 enzyme is identified by the UniProt reference P51648, a homolog, or a functional fragment thereof. In embodiments, the ALDH2 enzyme is identified by the UniProt reference P48448, a homolog, or a functional fragment thereof. In embodiments, the ALDH2 enzyme is identified by the UniProt reference S4R3S4, a homolog, or a functional fragment thereof.

[0023] “ALDH2*1” refers to the wild-type allele of the ALDH2 gene. The ALDH2*1 allele isassociated with normal ALDH2 enzyme activity, while the ALDH2*2 allele is a mutation that reduces or eliminates this activity.

[0024] “Aldehyde dehydrogenase 2 enzyme activity” or “ALDH2 enzyme activity” refers to catalyzing the chemical transformation from acetaldehyde to acetic acid in the major oxidative pathway of alcohol metabolism.

[0025] “Alcohol dehydrogenase 1 enzyme” or “ADH1 enzyme” or “Adh1 enzyme” includes any of the recombinant or naturally-occurring forms of the ADH1 enzyme or variants orhomologs thereof that maintain ADH1 enzyme activity (e.g., within at least 80%, at least 90%, at least 95%, or 100% activity compared to the ADH1 enzyme. In aspects, the variants or homologs have at least 90%, 95%, or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150, or 200 continuous amino acid portion) compared to the naturally occurring ADH1 enzyme. In embodiments, the ADH1 enzyme is identified by the UniProt reference P07327, a homolog, or a functional fragment thereof.

[0026] “ADH1B” or “ADH1B enzyme” or “alcohol dehydrogenase 1B enzyme” refers to a type of of ADH1 enzyme. For example, ADH1 enzyme is encompasses all class I alcohol dehydrogenase while ADH1B falls within the category of ADH1 enzymes. ADH1B encodes the beta subunit of class I alcohol dehydrogenase. In instances, the terms ADH1 and ADH1B may be used interchangeably.

[0027] “ADH1B*2” refers to the ADH1B2 allele of the ADH1B gene.

[0028] “Alcohol dehydrogenase 1 enzyme activity” or “ADH1 enzyme activity” refers to catalyzing the oxidation of primary and secondary alcohols to aldehydes and ketones, respectively.

[0029] “Lipid nanoparticle” refers to a lipid formulation that can be used to deliver an active agent or therapeutic agent, such as a nucleic acid (e.g., DNA, RNA, aRNA, siRNA, mRNA), to a target site of interest (e.g., cell, tissue, organ, and the like). In embodiments, the lipid nanoparticle is typically formed from a cationic lipid, a non-cationic lipid, and a conjugated lipid that prevents aggregation of the particle. In other embodiments, the active agent or therapeutic agent, such as a nucleic acid (e.g., DNA, RNA, aRNA, siRNA, mRNA), is encapsulated within the lipid nanoparticle, thereby protecting it from enzymatic degradation.

[0030] “Lipid” refers to a group of organic compounds that include, but are not limited to, esters of fatty acids. and are characterized by being insoluble in water, but soluble in many organic solvents.

[0031] “Lipid conjugate” refers to a conjugated lipid that inhibits aggregation of lipid particles. Such lipid conjugates include PEG-lipid conjugates such as, e.g., PEG conjugated to dialkyloxypropyls (e.g., DAA-PEG conjugates), PEG conjugated to diacylglycerols (e.g., DAG- PEG conjugates), PEG conjugated to cholesterol, PEG conjugated to phosphatidylethanol amines, PEG conjugated to ceramides, cationic PEG lipids, polyoxazoline (POZ)-lipid conjugates (e.g., POZ-DAA conjugates), polyamide oligomers (e.g., ATTA-lipid conjugates), and mixtures thereof. PEG or POZ can be conjugated directly to the lipid or may be linked to the lipid via a linker moiety. Any linker moiety suitable for coupling the PEG or the POZ to a lipidcan be used including, e.g., non-ester containing linker moieties and ester-containing linker moieties. In embodiments, non-ester containing linker moieties, such as amides or carbamates, are used.

[0032] “Polyethylene glycol-lipid conjugate” or “PEG-lipid conjugate” refers to a polyethylene glycol (PEG) having an average molecular weight from about 500 Daltons to about 10,000 Daltons conjugated to a C12-C22 fatty acid lipid. In embodiments, a PEG-lipid conjugate is polyethylene glycol having an average molecular weight from about 1,000 Daltons to about 6,000 Daltons conjugated to a C12-C22 fatty acid lipid. In embodiments, a PEG-lipid conjugate is polyethylene glycol having an average molecular weight from about 2,000 Daltons to about 5,000 Daltons conjugated to a C12-C20 fatty acid lipid. In embodiments, a PEG-lipid conjugate is polyethylene glycol having an average molecular weight from about 2,000 Daltons to about 5,000 Daltons conjugated to a C12-C18 fatty acid lipid. In embodiments, a PEG-lipid conjugate is polyethylene glycol having an average molecular weight from about 1,500 Daltons to about 2,500 Daltons conjugated to a C12-C18 fatty acid lipid. In embodiments, a PEG-lipid conjugate is polyethylene glycol having an average molecular weight from about 2,000 Daltons conjugated to a C12-C18 fatty acid lipid. In embodiments, the PEG-lipid conjugate is N-palmitoyl- sphingosine-1-{succinyl[methoxy-(polyethylene glycol)]} (C16 PEG ceramide), 1,2- dimyristoyl-rac-glycero-3-methoxy-polyethylene glycol (DMG-PEG), 1,2-dipalmitoyl-sn- glycero-3-phosphoethanolamine-N-[azido(polyethylene glycol) (DPPE-PEG), 1,2-dipalmitoyl- rac-glycero-3-methylpolyoxy-ethylene (DPG-PEG), distearoyl-rac-glycerol(polyethylene glycol) (DSG-PEG), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol) (DSPE-PEG). In embodiments, the polyethylene glycol has an average molecular weight of about 2000 daltons (e.g., DMG-PEG2000, DPPE-PEG2000, DPG-PEG2000, DSG-PEG2000, DSPE-PEG2000). In embodiments, DMG-PEG is a mixture of 1,2-DMG PEG2000 and 1,3- DMG PEG2000. In embodiments, DMG-PEG is a mixture of 1,2-DMG PEG2000 and 1,3-DMG PEG2000 in a ratio of about 99:1 to about 90:10. The polyethylene glycol-lipid conjugate can optionally be in the form of a pharmaceutically acceptable salt (e.g., ammonium salt).

[0033] “Average molecular weight” refers to the average molecular weight of a polymer sample that is determined by a technique known in the art, such as ultracentrifugation, gel permeation chromatography, light-scattering measurements, MALDI-TOF-Mass Spectrometry, USP 40-NF 35, and viscosity measurements. In embodiments, the average molecular weight is measured by MALDI-TOF-MS. In embodiments, the average molecular weight is measured by USP 40-NF 35. In embodiments, the average molecular weight is the number average molecular weight which is defined as the total weight of polymer divided by the total number of molecules.

[0034] “Amphipathic lipid” refers, in part, to any material wherein the hydrophobic portion of the lipid orients into a hydrophobic phase, while the hydrophilic portion orients toward the aqueous phase. Hydrophilic characteristics derive from the presence of polar or charged groups such as carbohydrates, phosphate, carboxylic, sulfato, amino, sulfhydryl, nitro, hydroxyl, and other like groups. Hydrophobicity can be conferred by the inclusion of apolar groups that include long-chain saturated and unsaturated aliphatic hydrocarbon groups and such groups substituted by one or more aromatic, cycloaliphatic, or heterocyclic group(s). Examples of amphipathic compounds include phospholipids, aminolipids, and sphingolipids. Other compounds lacking in phosphorus, such as sphingolipid, glycosphingolipid families, diacylglycerols, and β-acyloxyacids, are also within the group designated as amphipathic lipids. Additionally, the amphipathic lipids can be mixed with other lipids including triglycerides and sterols.

[0035] “Phospholipids” are a class of lipids whose molecule has a hydrophilic ”head” containing a phosphate group and two hydrophobic ”tails” derived from fatty acids, joined by an alcohol residue. Exemplary phospholipids include dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylethanolamine (DOPE), 1,2-di(9Z- octadecenoyl)-sn-glycero-3-phosphocholine (DOPC), palmitoyloleoyl-phosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), palmitoyloleyol- phosphatidylglycerol (POPG), dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoyl- phosphatidylethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), monomethyl-phosphatidylethanolamine (MMPE), dimethyl-phosphatidylethanolamine (DMPE), dielaidoyl-phosphatidylethanolamine (DEPE), stearoyloleoyl-phosphatidyl-ethanolamine (SOPE), egg phosphatidylcholine (EPC), hydrogenated soy phosphatidylcholine (HSPC), dipalmitoyl-phosphatidylglycerol (DPPG), and mixtures thereof. In embodiments, DSPC is 1,2- distearoyl-sn-glycero-3-phosphocholine. In embodiments, DPPG is 1,2-dipalmitoyl- phosphatidyl-glycerol. In embodiments, DPPG is 1,2-dihexadecanoyl-sn-glycero-3-phospho-(1'- sn-glycerol).

[0036] “Neutral lipid” refers to any of a number of lipid species that exist either in an uncharged or neutral zwitterionic form at a selected pH. At physiological pH, such lipids include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides, and diacylglycerols.

[0037] “Non-cationic lipid” refers to any amphipathic lipid as well as any other neutral lipid or anionic lipid.

[0038] “Anionic lipid” refers to any lipid that is negatively charged at physiological pH. Theselipids include, but are not limited to, phosphatidylglycerols, cardiolipins, diacylphosphatidylserines, diacylphosphatidic acids, N-dodecanoyl phosphatidylethanolamines, N-succinyl phosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids.

[0039] “Hydrophobic lipid” refers to compounds having apolar groups that include long-chain saturated and unsaturated aliphatic hydrocarbon groups and such groups optionally substituted by one or more aromatic, cycloaliphatic, or heterocyclic group(s). Examples include diacylglycerol, dialkylglycerol, N-N-dialkylamino, 1,2-diacyloxy-3-aminopropane, and 1,2- dialkyl-3-aminopropane.

[0040] A “cationic lipid” refers to a lipid capable of being positively charged or having a positive charge. Exemplary cationic lipids include an amine group which has a positive charge. Cationic lipids can be ionizable such that they can exist in a positively charged or neutral form depending on pH. The ionization of the cationic lipid affects the surface charge of the lipid nanoparticle under different pH conditions. The term “ionizable lipid” refers to a type of cationic lipid which is protonated at low pH (i.e., making them positively charged), but which remains neutral at physiological pH. Exemplary cationic lipids include MC3, MC3 derivatives, 1,2- dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 2,2-dilinoleyl-4-(2-dimethyl- aminoethyl)-[1,3]-dioxolane (DLin-KC2-DMA), 2,2-dilinoleyl-4-(3-dimethylaminopropyl)- [1,3]-dioxolane (DLin-K-3-DMA), 2,2-dilinoleyl-4-(4-dimethylaminobutyl)-[1,3]-dioxolane (DLin-KC4-DMA), 2,2-dilinoleyl-5-dimethylaminomethyl-[1,3]-dioxane (DLin-K6-DMA), 2,2- dilinoleyl-4-N-methylpepiazino-[1,3]-dioxolane (DLin-K-MPZ), 2,2-dilinoleyl-4-dimethyl- aminomethyl-[1,3]-dioxolane (DLin-K-DMA), 1,2-dilinoleylcarbamoyloxy-3-dimethyl- aminopropane (DLin-C-DAP), 1,2-dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin- DAC), 1,2-dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethyl- aminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1- linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy-3- trimethylaminopropane or a salt thereof (DLin-TMA), 1,2-dilinoleoyl-3-trimethylaminopropane or a salt thereof (DLin-TAP), 1,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), 3- (N,N-dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleylamino)-1,2-propanedio (DOAP), 1,2-dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 1,2- N,N′-dilinoleylcarbamyl-3-dimethylaminopropane (DLincarbDAP), N,N-dioleyl-N,N- dimethylammonium chloride (DODAC), 1,2-dioleyloxy-N,N-dimethyl-aminopropane (DODMA), 1,2-distearyloxy-N,N-dimethylaminopropane (DSDMA), N-(1-(2,3-dioleyloxy)-propyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-distearyl-N,N-dimethyl- ammonium bromide (DDAB), 1,2-dioleoyloxy-3-(trimethylammonium)propane (DOTAP), 3- (N-(N′,N′-dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(1,2-dimyristyloxyprop- 3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (DMRIE), 2,3-dioleyloxy-N- [2(spermine-carboxamido)ethyl]-N,N-dimethyl-1-propanaminium-trifluoroacetate (DOSPA), dioctadecylamidoglycyl spermine (DOGS), 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutan- 4-oxy)-1-(cis,cis-9,12-octadecadienoxy)-propane (CLinDMA), 2-[5′-(cholest-5-en-3-beta-oxy)- 3′-oxapentoxy)-3-dimethy-1-(cis,cis-9′,1-2′-octadecadienoxy)-propane (CpLinDMA), N,N- dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N′-dioleylcarbamyl-3-dimethyl- aminopropane (DOcarbDAP), 3-(N,N-dioleylamino)-1,2-propanedio (DOAP), and 1,1’-((2-(4- (2-((2-(bis(2-hydroxydodecyl)amino)ethyl) (C12-200), N1,N16-didodecyl-4,7,13-tris[3- (dodecylamino)-3-oxopropyl]-4,7,10,13-tetraazahexadecanediamide (98N12-5), heptadecan-9-yl 8-((2-hydroxyethyl)(8-(nonyloxy)-8-oxooctyl)amino)octanoate (Lipid 5), Lipid A9, 4A3-SC8, tetrakis(2-(octyldisulfaneyl)ethyl) 3,3',3'',3'''-(((methylazanediyl)bis(propane-3,1-diyl))bis- (azanetriyl))tetrapropionate (306-012B), 1,2-dioleoyl-3-dimethylammonium propane (DODAP), 9-heptadecanyl 8-{(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino}octanoate (SM-102), 6- ((2-hexyldecanoyl)oxy)-N-(6-((2-hexyldecanoyl)oxy)hexyl)-N-(4-hydroxybutyl)hexan-1- aminium (ALC-0315), 9-[4-(dimethylamino)-1-oxobutoxy]-heptadecanedioic acid, 1,17-di-(2Z)- 2-nonen-1-yl ester (Lipid 319 or L-319), 5A2-SC8, tetrakis(8-methylnonyl) 3,3',3'',3'''- (((methylazanediyl)bis(propane-3,1-diyl))bis(azanetriyl))tetrapropionate (306Oi10), 5- (dimethylamino)-pentanoic acid, (6Z)-1,2-di-(4Z)-4-decen-1-yl-6-dodecen-1-yl ester (Lipid CL1), and [(Z)-non-2-enyl] 2-[4-(dimethylamino)butylsulfanylcarbonyl-(2-heptadecan-9-yloxy- 2-oxoethyl)amino]acetate (Arcturus Lipid 2,2 or ATX-0114).

[0041] “Lipid A9” refers to the compound having the following structure, as described in Buschmann et al, Vaccines, 9, 65 (2021); doi.org / 10.3390 / vaccines9010065: .

[0042] in Wang et al, Nature Protocols, 18:265-291 (2023):. te. In embodiments, “MC3” refers to heptatriaconta-6,9,28,31-tetraen-19-yl 4- (dimethylamino)butanoate. In embodiments, “MC3” refers to (6Z,9Z,28Z,31Z)-heptatriaconta- 6,9,28,31-tetraen-19-yl-4-(dimethylamino)butanoate.

[0044] “MC3 derivative” refers to derivatives of MC3 such as those described in US Publication No.2017 / 0151333. Exemplary MC3 derivatives include LenMC3, γ-LenMC3, MC3 ether, MC4 ether, MC3MC, MC2C, MC2MC, MC3 thioester, MC3 alkyne, and MC3 amide.

[0045] “Non-lamellar morphology” refer to a non-bilayer structure. The non-bilayer morphology can include, for example, three dimensional tubes, rods, cubic symmetries, etc. The non-lamellar morphology (i.e., non-bilayer structure) of the lipid particles can be determined using analytical techniques including Cryo-Transmission Electron Microscopy (“Cryo-TEM”), Differential Scanning calorimetry (“DSC”), and X-Ray Diffraction.

[0046] The singular terms “a,” “an,” and “the” include the plural reference unless the context clearly indicates otherwise.

[0047] A “plurality of lipid nanoparticles” refers to at least 2 lipid nanoparticles, more preferably more than 102, 103, 104, 105, 106or more lipid nanoparticles (or any fraction thereof or range therein). In embodiments, the plurality of lipid nanoparticles includes 50-100, 50-200, 50-300, 50-400, 50-500, 50-600, 50-700, 50-800, 50-900, 50-1000, 50-1100, 50-1200, 50-1300, 50-1400, 50-1500, 50-1600, 50-1700, 50-1800, 50-1900, 50-2000, 50-2500, 50-3000, 50-3500, 50-4000, 50-4500, 50-5000, 50-5500, 50-6000, 50-6500, 50-7000, 50-7500, 50-8000, 50-8500, 50-9000, 50-9500, 50-10,000 or more lipid nanoparticles.

[0048] “Nucleic acid” refers to nucleotides (e.g., deoxyribonucleotides or ribonucleotides) and polymers thereof in either single-, double- or multiple-stranded form, or complements thereof. The terms “polynucleotide,” “oligonucleotide,” “oligo” or the like refer, in the usual and customary sense, to a linear sequence of nucleotides. The term “nucleotide” refers, in the usual and customary sense, to a single unit of a polynucleotide, i.e., a monomer. Nucleotides can be ribonucleotides, deoxyribonucleotides, or modified versions thereof. Examples of nucleic acids contemplated herein include single and double stranded DNA, single and double stranded RNA, and hybrid molecules having mixtures of single and double stranded DNA and RNA. Examplesof nucleic acids contemplated herein include any types of RNA (e.g., antisense RNA, mRNA, siRNA, miRNA, shRNA, guide RNA, dicer substrate RNA, dicer substrate siRNAs (dsiRNAs) (dsiRNA are cleaved by the RNase III class endoribonuclease dicer into 21-23 base duplexes having 2-base 3’-overhangs siRNA), and any type of DNA, genomic DNA, plasmid DNA, minicircle DNA, minigene, and any fragments thereof. The terms “messenger RNA” or “mRNA” refer a single-stranded molecule of RNA that corresponds to the genetic sequence of a gene, and is read by a ribosome in the process of synthesizing a protein. Nucleic acids can be linear or branched. For example, nucleic acids can be a linear chain of nucleotides or the nucleic acids can be branched, e.g., such that the nucleic acids comprise one or more arms or branches of nucleotides. Optionally, the branched nucleic acids are repetitively branched to form higher ordered structures such as dendrimers and the like.

[0049] The terms also encompass nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non- naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, include, without limitation, phosphodiester derivatives including, e.g., phosphoramidate, phosphorodiamidate, phosphorothioate (also known as phosphorothioate having double bonded sulfur replacing oxygen in the phosphate), phosphorodithioate, phosphonocarboxylic acids, phosphonocarboxylates, phosphonoacetic acid, phosphonoformic acid, methyl phosphonate, boron phosphonate, or O-methylphosphoroamidite linkages (see Eckstein, Oligonucleotides and Analogues: A Practical Approach, Oxford University Press) as well as modifications to the nucleotide bases such as 2’O-methyl, 2’O-methoxyethoxy, 2’fluoro, 5-methyl cytidine or pseudouridine; and peptide nucleic acid backbones and linkages. Other analog nucleic acids include those with positive backbones; non-ionic backbones, modified sugars (e.g., deoxyribose), and non-ribose backbones (e.g. phosphorodiamidate morpholino oligos or locked nucleic acids (LNA) as known in the art), including those described in U.S. Patent Nos.5,235,033 and 5,034,506, and Chapters 6 and 7, ASC Symposium Series 580, Carbohydrate Modifications in Antisense Research, Sanghui & Cook, eds. Nucleic acids containing one or more carbocyclic sugars are also included within one definition of nucleic acids. Modifications of the ribose-phosphate backbone may be done for a variety of reasons, e.g., to increase the stability and half-life of such molecules in physiological environments or as probes on a biochip. Mixtures of naturally occurring nucleic acids and analogs can be made; alternatively, mixtures of different nucleic acid analogs, and mixtures of naturally occurring nucleic acids and analogs may be made. In embodiments, the internucleotide linkages in DNAare phosphodiester, phosphodiester derivatives, or a combination of both.

[0050] Nucleic acids, including, e.g., nucleic acids with a phosphothioate backbone, can include one or more reactive moieties. As used herein, the term reactive moiety includes any group capable of reacting with another molecule, e.g., a nucleic acid or polypeptide through covalent, non-covalent or other interactions. By way of example, the nucleic acid can include an amino acid reactive moiety that reacts with an amio acid on a protein or polypeptide through a covalent, non-covalent or other interaction.

[0051] Nucleic acids can include nonspecific sequences. As used herein, the term “nonspecific sequence” refers to a nucleic acid sequence that contains a series of residues that are not designed to be complementary to or are only partially complementary to any other nucleic acid sequence. By way of example, a nonspecific nucleic acid sequence is a sequence of nucleic acid residues that does not function as an inhibitory nucleic acid when contacted with a cell or organism.

[0052] “Hybridize” and “hybridization” refer to the pairing of complementary (including partially complementary) nucleic acid strands. Hybridization and the strength of hybridization (e.g., the strength of the association between nucleic acid strands) is impacted by factors known in the art including the degree of complementarity between the nucleic acid, stringency of the conditions involved affected by such conditions as the concentration of salts, the melting temperature (Tm) of the formed hybrid, the presence of other components, the molarity of the hybridizing strands and the G:C content of the nucleic acid strands. When one nucleic acid is said to “hybridize” to another nucleic acid, it means that there is some complementarity between the two nucleic acids or that the two nucleic acids form a hybrid under high or low stringency conditions.

[0053] “Complement” refers to a nucleotide (e.g., RNA or DNA) or a sequence of nucleotides capable of base pairing with a complementary nucleotide or sequence of nucleotides. As described herein and commonly known in the art the complementary (matching) nucleotide of adenosine is thymidine and the complementary (matching) nucleotide of guanidine is cytosine. Thus, a complement may include a sequence of nucleotides that base pair with corresponding complementary nucleotides of a second nucleic acid sequence. The nucleotides of a complement may partially or completely match the nucleotides of the second nucleic acid sequence. Where the nucleotides of the complement completely match each nucleotide of the second nucleic acid sequence, the complement forms base pairs with each nucleotide of the second nucleic acid sequence. Where the nucleotides of the complement partially match the nucleotides of the second nucleic acid sequence only some of the nucleotides of the complementform base pairs with nucleotides of the second nucleic acid sequence. Examples of complementary sequences include coding and a non-coding sequences, wherein the non-coding sequence contains complementary nucleotides to the coding sequence and thus forms the complement of the coding sequence. A further example of complementary sequences are sense and antisense sequences, wherein the sense sequence contains complementary nucleotides to the antisense sequence and thus forms the complement of the antisense sequence. The complementarity of sequences may be partial, in which only some of the nucleic acids match according to base pairing, or complete, where all the nucleic acids match according to base pairing. Thus, two sequences that are complementary to each other, may have a specified percentage of nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region).

[0054] “Hybridization conditions” refers to conditions under which a nucleic acid will hybridize to its target sequence, typically in a complex mixture of nucleic acids, but to no other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures. An extensive guide to the hybridization of nucleic acids is found in Tijssen, Techniques in Biochemistry and Molecular Biology-Hybridization with Nucleic Probes, “Overview of principles of hybridization and the strategy of nucleic acid assays” (1993). Generally, stringent conditions are selected to be about 5-10°C. lower than thermal melting point (Tm) for the specific sequence at a defined ionic strength pH. The Tmis the temperature (under defined ionic strength, pH, and nucleic concentration) at which 50% of the probes complementary to the target hybridize to the target sequence at equilibrium (as the target sequences are present in excess, at Tm, 50% of the probes are occupied at equilibrium). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. For selective or specific hybridization, a positive signal is at least two times background, preferably 10 times background hybridization. Exemplary hybridization conditions can be as follows: 50% formamide, 5×SSC, and 1% SDS, incubating at 42°C, or 5×SSC, 1% SDS, incubating at 65°C, with wash in 0.2×SSC, and 0.1% SDS at 65°C. For PCR, a temperature of about 36°C is typical for low stringency amplification, although annealing temperatures may vary between about 32°C and 48°C depending on primer length. For PCR amplification, a temperature of about 62°C is typical, although high stringency annealing temperatures can range from about 50°C to about 65°C depending on the primer length and specificity. Typical cycle conditions for both high and low stringency amplifications include a denaturation phase of 90°C-95°C for 30 seconds to 2 minutes, an annealing phaselasting 30 seconds to 2 minutes, and an extension phase of about 72°C for 1-2 min. Protocols and guidelines for low and high stringency amplification reactions are provided, e.g., Innis et al., PCR Protocols, A Guide to Methods and Applications, Academic Press, Inc. N.Y. (1990).

[0055] A polynucleotide is typically composed of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T) (uracil (U) for thymine (T) when the polynucleotide is RNA). Thus, the term “polynucleotide sequence” is the alphabetical representation of a polynucleotide molecule; alternatively, the term may be applied to the polynucleotide molecule itself. This alphabetical representation can be input into databases in a computer having a central processing unit and used for bioinformatics applications such as functional genomics and homology searching. Polynucleotides may optionally include one or more non-standard nucleotide(s), nucleotide analog(s) and / or modified nucleotides.

[0056] “Conservatively modified variants” applies to both amino acid and nucleic acidsequences. With respect to particular nucleic acid sequences, “conservatively modified variants” refers to those nucleic acids that encode identical or essentially identical amino acid sequences. Because of the degeneracy of the genetic code, a number of nucleic acid sequences will encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are “silent variations,” which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. One of skill will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence.

[0057] “Identical” or percent “identity,” in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (e.g., ncbi.nlm.nih.gov / BLAST / or the like). Such sequences are then said to be “substantially identical.” This definition also refers to, or may beapplied to, the compliment of a test sequence. The definition also includes sequences that have deletions and / or additions, as well as those that have substitutions. The preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 amino acids or nucleotides in length. In embodiments, the percentage of sequence identity is determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.

[0058] An amino acid or nucleotide base “position” is denoted by a number that sequentially identifies each amino acid (or nucleotide base) in the reference sequence based on its position relative to the N-terminus (or 5'-end). Due to deletions, insertions, truncations, fusions, and the like that must be taken into account when determining an optimal alignment, in general the amino acid residue number in a test sequence determined by simply counting from the N- terminus will not necessarily be the same as the number of its corresponding position in the reference sequence. For example, in a case where a variant has a deletion relative to an aligned reference sequence, there will be no amino acid in the variant that corresponds to a position in the reference sequence at the site of deletion. Where there is an insertion in an aligned reference sequence, that insertion will not correspond to a numbered amino acid position in the reference sequence. In the case of truncations or fusions there can be stretches of amino acids in either the reference or aligned sequence that do not correspond to any amino acid in the corresponding sequence.

[0059] “Numbered with reference to” or “corresponding to,” when used in the context of the numbering of a given amino acid or polynucleotide sequence, refers to the numbering of the residues of a specified reference sequence when the given amino acid or polynucleotide sequence is compared to the reference sequence.

[0060] “Isolated” when applied to a nucleic acid or protein denotes that the nucleic acid or protein is essentially free of other cellular components with which it is associated in the natural state. It can be, for example, in a homogeneous state and may be in either a dry or aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniquessuch as polyacrylamide gel electrophoresis or high performance liquid chromatography. A protein that is the predominant species present in a preparation is substantially purified. In embodiments, the nucleic acids described herein are isolated nucleic acids.

[0061] “Expression” includes any step involved in the production of the polypeptide including, but not limited to, transcription, post-transcriptional modification, translation, post- translational modification, and secretion. Expression can be detected using conventional techniques for detecting protein (e.g., ELISA, Western blotting, flow cytometry, immunofluorescence, immunohistochemistry, etc.).

[0062] “Pharmaceutically acceptable salts” refers to salts of the active compounds that are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. Any of the compounds described herein can be in the form of a pharmaceutically acceptable salt. When compounds of the present disclosure contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present disclosure contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, phosphoric, sulfuric, monohydrogencarbonic, monohydrogenphosphoric, dihydrogenphosphoric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, oxalic, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1- 19). Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0063] “Treating” or “treatment” are used in accordance with their plain and ordinary meaning and broadly includes any approach for obtaining beneficial or desired results in a subject’s condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment ofthe extent of a disease, stabilizing (i.e., not worsening) the state of disease, delay or slowing of disease progression, amelioration of the disease state, and remission, whether partial or total and whether detectable or undetectable. Treatment may inhibit the disease’s spread; relieve the disease’s symptoms, fully or partially remove the disease’s underlying cause, shorten a disease’s duration, or do a combination of these things. Treatment methods include administering to a subject a therapeutically effective amount of an active agent. The term “treating” does not including preventing.

[0064] “Patient” or “subject” refers to a living organism. Non-limiting examples include humans, other mammals, dogs, cats, rats, mice, monkeys, and non-mammalian animals. In embodiments, a patient is a mammal. In embodiments, a patient is human.

[0065] “Supplemental magnesium” refers to Mg2+in any form, such as pharmaceutically acceptable salts of magnesium, including magnesium sulfate, magnesium citrate, magnesium oxide, magnesium chloride, magnesium carbonate, magnesium lactate, magnesium malate, magnesium gluconate, magnesium hydroxide, magnesium glycinate, or chelated magnesium. Supplemental magnesium can be administered as a solid or liquid (e.g., magnesium sulfate in water). Supplemental magnesium can be administered orally or parenterally (e.g., intravenous, intramuscular).

[0066] “Supplemental zinc” refers to Zn2” in any form, such as pharmaceutically acceptable salts of zinc, including zinc sulfate, zinc acetate, zinc gluconate, zinc glycinate, zinc picolinate, zinc citrate, zinc oxide, or chelated zinc. Supplemental zinc can be administered as a solid or liquid. Supplemental zinc can be administered orally or parenterally.

[0067] “Vitamin B3” is a vitamin family that includes niacin (or nicotinic acid), nicotinamide (or niacinamide), and nicotinamide riboside. Niacin is C6H5NO2. Nicotinamide is C6H6N2O. Nicotinamide riboside is C11H15N2O5+.

[0068] A “effective amount” as used herein, is an amount sufficient for a compound to accomplish a stated purpose relative to the absence of the compound (e.g. achieve the effect for which it is administered, treat a disease, reduce enzyme activity, increase enzyme activity, reduce a signaling pathway, or reduce one or more symptoms of a disease or condition). In these methods, the effective amount of the nucleic acid (DNA, RNA, mRNA) described herein is an amount effective to accomplish the stated purpose of the method. An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.” A “reduction” of a symptom or symptoms (and grammatical equivalents ofthis phrase) means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s). The exact amounts will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols.1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0069] “Therapeutically effective amount,” as used herein, refers to that amount of therapeutic agent sufficient to ameliorate the disorder, as described above. For example, for the given parameter, a therapeutically effective amount will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy can also be expressed as “-fold” increase or decrease. For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more effect over a control. For any compound described herein, therapeutically effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of active compound(s) that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art. As is in the art, therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals.

[0070] For any therapeutic agent (e.g., nucleic acid) described herein, the therapeutically effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of active compound that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art. Therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring compounds effectiveness and adjusting the dosage upwards or downwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods is well within the capabilities of the ordinarily skilled artisan.

[0071] “Administering” means intranasal administration, inhalation administration, oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal,sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. In embodiments, administering does not include administration of any active agent other than the nucleic acid. In embodiments, administration is intranasal. In embodiments, administration is intravenous. In embodiments, administration is intranasal administration of lipid nanoparticles. In embodiments, administration is intravenous administration of lipid nanoparticles.

[0072] “About” means a range of values including the specified value, which a person of ordinary skill in the art would consider reasonably similar to the specified value. In embodiments, “about” means within a standard deviation using measurements generally acceptable in the art. In embodiments, “about” means a range extending to + / - 15% of the specified value. In embodiments, “about” means a range extending to + / - 10% of the specified value. In embodiments, “about” means a range extending to + / - 5% of the specified value.

[0073] Nucleic Acids

[0074] Provided herein are nucleic acids. In embodiments, the nucleic acid encodes an ALDH2 enzyme. In embodiments, the nucleic acid encodes an ADH1 enzyme. In embodiments, the nucleic acid encodes an ALDH2 enzyme and an ADH1 enzyme.

[0075] In embodiments, the ALDH2 enzyme has at least 85% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 90% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 91% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 2% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 93% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 94% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 95% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 96% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 97% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 98% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has at least 99% sequence identity to SEQ ID NO:7. In embodiments, the ALDH2 enzyme has SEQ ID NO:7.

[0076] In embodiments, the ALDH2 enzyme has at least 85% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 90% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 91% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 2% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 93% sequence identity to SEQ IDNO:11. In embodiments, the ALDH2 enzyme has at least 94% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 95% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 96% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 97% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 98% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has at least 99% sequence identity to SEQ ID NO:11. In embodiments, the ALDH2 enzyme has SEQ ID NO:11. In embodiments of SEQ ID NO:11, X4is E. In embodiments, X4is S. In embodiments, X4is N. In embodiments, X4is D. In embodiments, X4is A. In embodiments, X4is R. In embodiments, X4is C. In embodiments, X4is Q. In embodiments, X4is G. In embodiments, X4is P. In embodiments, X4is Y. In embodiments, X4is H. In embodiments, X4is I. In embodiments, X4is L. In embodiments, X4is K. In embodiments, X4is M. In embodiments, X4is F. In embodiments, X4is T. In embodiments, X4is W. In embodiments, X4is V. In embodiments where the ALDH2 enzyme has less than 100% sequence identity to SEQ ID NO:11, the amino acid X4is present at the position corresponding to X4in SEQ ID NO:11.

[0077] In embodiments, the ALDH2 enzyme has at least 85% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 90% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 91% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 2% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 93% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 94% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 95% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 96% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 97% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 98% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has at least 99% sequence identity to SEQ ID NO:14. In embodiments, the ALDH2 enzyme has SEQ ID NO:14. In embodiments of SEQ ID NO:14, X5is E. In embodiments, X5is S. In embodiments, X5is N. In embodiments, X5is D. In embodiments, X5is A. In embodiments, X5is R. In embodiments, X5is C. In embodiments, X5is Q. In embodiments, X5is G. In embodiments, X5is P. In embodiments, X5is Y. In embodiments, X5is H. In embodiments, X5is I. In embodiments, X5is L. In embodiments, X5is K. In embodiments, X5is M. In embodiments, X5is F. In embodiments, X5is T. In embodiments, X5is W. In embodiments, X5is V. In embodiments where the ALDH2 enzyme has less than 100% sequence identity to SEQ ID NO:14, the amino acid X5is present at theposition corresponding to X5in SEQ ID NO:14.

[0078] In embodiments, the ADH1 enzyme has at least 85% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 90% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 91% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 92% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 93% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 94% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 95% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 96% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 97% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 98% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has at least 99% sequence identity to SEQ ID NO:8. In embodiments, the ADH1 enzyme has SEQ ID NO:8.

[0079] In embodiments, the ADH1 enzyme has at least 85% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 90% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 91% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 92% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 93% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 94% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 95% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 96% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 97% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 98% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has at least 99% sequence identity to SEQ ID NO:17. In embodiments, the ADH1 enzyme has SEQ ID NO:17. In embodiments of SEQ ID NO:17, X6is E. In embodiments, X6is S. In embodiments, X6is N. In embodiments, X6is D. In embodiments, X6is A. In embodiments, X6is R. In embodiments, X6is C. In embodiments, X6is Q. In embodiments, X6is G. In embodiments, X6is P. In embodiments, X6is Y. In embodiments, X6is H. In embodiments, X6is I. In embodiments, X6is L. In embodiments, X6is K. In embodiments, X6is M. In embodiments, X6is F. In embodiments, X6is T. In embodiments, X6is W. In embodiments, X6is V. In embodiments where the ADH1 enzyme has less than 100% sequence identity to SEQ ID NO:17, the amino acid X6is present at the position corresponding to X6in SEQ ID NO:17.

[0080] In embodiments, the ADH1 enzyme has at least 85% sequence identity to SEQ IDNO:20. In embodiments, the ADH1 enzyme has at least 90% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 91% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 92% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 93% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 94% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 95% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 96% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 97% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 98% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has at least 99% sequence identity to SEQ ID NO:20. In embodiments, the ADH1 enzyme has SEQ ID NO:20. In embodiments of SEQ ID NO:20, X7is E. In embodiments, X7is S. In embodiments, X7is N. In embodiments, X7is D. In embodiments, X7is A. In embodiments, X7is R. In embodiments, X7is C. In embodiments, X7is Q. In embodiments, X7is G. In embodiments, X7is P. In embodiments, X7is Y. In embodiments, X7is H. In embodiments, X7is I. In embodiments, X7is L. In embodiments, X7is K. In embodiments, X7is M. In embodiments, X7is F. In embodiments, X7is T. In embodiments, X7is W. In embodiments, X7is V. In embodiments of SEQ ID NO:20, X8is E. In embodiments, X8is S. In embodiments, X8is N. In embodiments, X8is D. In embodiments, X8is A. In embodiments, X8is R. In embodiments, X8is C. In embodiments, X8is Q. In embodiments, X8is G. In embodiments, X8is P. In embodiments, X8is Y. In embodiments, X8is H. In embodiments, X8is I. In embodiments, X8is L. In embodiments, X8is K. In embodiments, X8is M. In embodiments, X8is F. In embodiments, X8is T. In embodiments, X8is W. In embodiments, X8is V. In embodiments where the ADH1 enzyme has less than 100% sequence identity to SEQ ID NO:20, the amino acids X7and X8are present at the positions corresponding to X7and X8, respectively, in SEQ ID NO:20.

[0081] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has anucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:1. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:1. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0082] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:2. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:2. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0083] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90%sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:3. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:3. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0084] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:9. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:9. In embodiments of SEQ ID NO:9, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:9, the codon X1X2X3is present at the position corresponding to X1X2X3in SEQ ID NO:9. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0085] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid thatencodes the ALDH2 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:10. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:10. In embodiments of SEQ ID NO:10, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:10, the codon X1X2X3is present at the position corresponding to X1X2X3in SEQ ID NO:10. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0086] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid thatencodes the ALDH2 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:12. In embodiments of SEQ ID NO:12, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:12, the codon X1X2X3is present at the position corresponding to X1X2X3in SEQ ID NO:12. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0087] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid thatencodes the ALDH2 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:13. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:13. In embodiments of SEQ ID NO:13, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:13, the codon X1X2X3is present at the position corresponding to X1X2X3in SEQ ID NO:13. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0088] In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid thatencodes the ALDH2 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:21. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has SEQ ID NO:21.

[0089] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:4. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:4. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0090] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least95% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:5. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:5. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0091] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:6. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:6. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0092] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:15. Inembodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:15. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:15. In embodiments of SEQ ID NO:15, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:15, the codon X1X2X3is present at the position corresponding to X1X2X3in SEQ ID NO:15. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0093] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:16. Inembodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:16. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:16. In embodiments of SEQ ID NO:16, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:16, the codon X1X2X3is present at the position corresponding to X1X2X3in SEQ ID NO:16. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0094] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:18. Inembodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:18. In embodiments of SEQ ID NO:18, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments, XAXBXCis a codon that encodes E. In embodiments, XAXBXCis a codon that encodes S. In embodiments, XAXBXCis a codon that encodes N. In embodiments, XAXBXCis a codon that encodes D. In embodiments, XAXBXCis a codon that encodes A. In embodiments, XAXBXCis a codon that encodes R. In embodiments, XAXBXCis a codon that encodes C. In embodiments, XAXBXCis a codon that encodes Q. In embodiments, XAXBXCis a codon that encodes G. In embodiments, XAXBXCis a codon that encodes P. In embodiments, XAXBXCis a codon that encodes Y. In embodiments, XAXBXCis a codon that encodes H. In embodiments, XAXBXCis a codon that encodes I. In embodiments, XAXBXCis a codon that encodes L. In embodiments,XAXBXCis a codon that encodes K. In embodiments, XAXBXCis a codon that encodes M. In embodiments, XAXBXCis a codon that encodes F. In embodiments, XAXBXCis a codon that encodes T. In embodiments, XAXBXCis a codon that encodes W. In embodiments, XAXBXCis a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:18, the codons X1X2X3and XAXBXCare present at the positions corresponding to X1X2X3and XAXBXC, respectively, in SEQ ID NO:18. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0095] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:19. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:19. In embodiments of SEQ ID NO:19, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments,X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments, XAXBXCis a codon that encodes E. In embodiments, XAXBXCis a codon that encodes S. In embodiments, XAXBXCis a codon that encodes N. In embodiments, XAXBXCis a codon that encodes D. In embodiments, XAXBXCis a codon that encodes A. In embodiments, XAXBXCis a codon that encodes R. In embodiments, XAXBXCis a codon that encodes C. In embodiments, XAXBXCis a codon that encodes Q. In embodiments, XAXBXCis a codon that encodes G. In embodiments, XAXBXCis a codon that encodes P. In embodiments, XAXBXCis a codon that encodes Y. In embodiments, XAXBXCis a codon that encodes H. In embodiments, XAXBXCis a codon that encodes I. In embodiments, XAXBXCis a codon that encodes L. In embodiments, XAXBXCis a codon that encodes K. In embodiments, XAXBXCis a codon that encodes M. In embodiments, XAXBXCis a codon that encodes F. In embodiments, XAXBXCis a codon that encodes T. In embodiments, XAXBXCis a codon that encodes W. In embodiments, XAXBXCis a codon that encodes V. In embodiments where the nucleic acid has less than 100% sequence identity to SEQ ID NO:19, the codons X1X2X3and XAXBXCare present at the positions corresponding to X1X2X3and XAXBXC, respectively, in SEQ ID NO:19. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0096] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:22. Inembodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:22. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:22.

[0097] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:23. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:23.

[0098] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 91% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 92% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 93% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has anucleic acid sequence that has at least 94% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 96% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 97% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 98% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 99% sequence identity to SEQ ID NO:24. In embodiments, the nucleic acid that encodes the ADH1 enzyme has SEQ ID NO:24.

[0099] Provided herein is a nucleic acid that encodes the ALDH2 enzyme that has a nucleic acid sequence that has at least 85% sequence identity to the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 85% sequence identity to the nucleic acid sequence beginning after the start codon to before the beginning of the stop codon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 90% sequence identity to the nucleic acid sequence beginning after the start codon to before the beginning of the stop codon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has a nucleic acid sequence that has at least 95% sequence identity to the nucleic acid sequence beginning after the start codon to before the beginning of the stop codon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid that encodes the ALDH2 enzyme has the nucleic acid sequence beginning after the start codon to before the beginning of the stopcodon of SEQ ID NO:1, SEQ ID NO:9, or SEQ ID NO:12. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA. Provided herein is an ALDH2 enzyme encoded by any of the nucleic acids described herein. The pharse “the beginning of the start codon” means nucleic acid sequence includes the start codon. The phrase “beginning after the start codon” means the nucleic acid sequence does not include the start codon. The phrase “before the beginning of the stop codon” means the nucleic acid sequence does not include the stop codon.

[0100] In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has the nucleic acid sequence spanning the beginning of the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 85% sequence identity to the nucleic acid sequence beginning after the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 90% sequence identity to the nucleic acid sequence beginning after the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has a nucleic acid sequence that has at least 95% sequence identity to the nucleic acid sequence beginning after the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid that encodes the ADH1 enzyme has the nucleic acid sequence beginning after the start codon to before the beginning of the stop codon of SEQ ID NO:4, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:18. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA. Provided herein is an ADH1 enzyme encoded by any of the nucleic acids describedherein. The pharse “the beginning of the start codon” means nucleic acid sequence includes the start codon. The phrase “beginning after the start codon” means the nucleic acid sequence does not include the start codon. The phrase “before the beginning of the stop codon” means the nucleic acid sequence does not include the stop codon.

[0101] In embodiments, the nucleic acid encodes an ALDH2 enzyme and an ADH1 enzyme. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 85% sequence identity to a nucleic acid comprising: (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 90% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 91% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 92% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, or SEQ ID NO:19. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 93% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 94% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 95% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 96% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ IDNO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 97% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, or SEQ ID NO:13, and (ii) SEQ ID NO:5, SEQ ID NO:16, or SEQ ID NO:19. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 98% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence with at least 99% sequence identity to a nucleic acid comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid that encodes an ALDH2 enzyme and an ADH1 enzyme has a nucleic acid sequence comprising (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21, and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24. In embodiments, the nucleic acid sequence comprises SEQ ID NO:2 (or a percent sequence identity thereof) and SEQ ID NO:5 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:2 (or a percent sequence identity thereof) and SEQ ID NO:16 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:2 (or a percent sequence identity thereof) and SEQ ID NO:19 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:10 (or a percent sequence identity thereof) and SEQ ID NO:5 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:10 (or a percent sequence identity thereof) and SEQ ID NO:16 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:10 (or a percent sequence identity thereof) and SEQ ID NO:19 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:13 (or a percent sequence identity thereof) and SEQ ID NO:5 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:13 (or a percent sequence identity thereof) and SEQ ID NO:16 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:13 (or a percent sequence identity thereof) and SEQ ID NO:19 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:21 (or a percent sequence identity thereof) and SEQ ID NO:5 (or a percent sequence identity thereof). Inembodiments, the nucleic acid sequence comprises SEQ ID NO:21 (or a percent sequence identity thereof) and SEQ ID NO:16 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:21 (or a percent sequence identity thereof) and SEQ ID NO:19 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:2 (or a percent sequence identity thereof) and SEQ ID NO:22 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:10 (or a percent sequence identity thereof) and SEQ ID NO:22 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:13 (or a percent sequence identity thereof) and SEQ ID NO:22 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:21 (or a percent sequence identity thereof) and SEQ ID NO:22 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:2 (or a percent sequence identity thereof) and SEQ ID NO:23 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:10 (or a percent sequence identity thereof) and SEQ ID NO:23 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:13 (or a percent sequence identity thereof) and SEQ ID NO:23 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:21 (or a percent sequence identity thereof) and SEQ ID NO:23 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:2 (or a percent sequence identity thereof) and SEQ ID NO:24 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:10 (or a percent sequence identity thereof) and SEQ ID NO:24 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:13 (or a percent sequence identity thereof) and SEQ ID NO:24 (or a percent sequence identity thereof). In embodiments, the nucleic acid sequence comprises SEQ ID NO:21 (or a percent sequence identity thereof) and SEQ ID NO:24 (or a percent sequence identity thereof). In embodiments of SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, and SEQ ID NO:19, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments,X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V. In embodiments of SEQ ID NO:19, XAXBXCis a codon that encodes E. In embodiments, XAXBXCis a codon that encodes S. In embodiments, XAXBXCis a codon that encodes N. In embodiments, XAXBXCis a codon that encodes D. In embodiments, XAXBXCis a codon that encodes A. In embodiments, XAXBXCis a codon that encodes R. In embodiments, XAXBXCis a codon that encodes C. In embodiments, XAXBXCis a codon that encodes Q. In embodiments, XAXBXCis a codon that encodes G. In embodiments, XAXBXCis a codon that encodes P. In embodiments, XAXBXCis a codon that encodes Y. In embodiments, XAXBXCis a codon that encodes H. In embodiments, XAXBXCis a codon that encodes I. In embodiments, XAXBXCis a codon that encodes L. In embodiments, XAXBXCis a codon that encodes K. In embodiments, XAXBXCis a codon that encodes M. In embodiments, XAXBXCis a codon that encodes F. In embodiments, XAXBXCis a codon that encodes T. In embodiments, XAXBXCis a codon that encodes W. In embodiments, XAXBXCis a codon that encodes V. In embodiments, the nucleic acid is RNA. In embodiments, the nucleic acid is mRNA.

[0102] Lipid Nanoparticles

[0103] Provided herein are lipid nanoparticles. In embodiments, the lipid nanoparticles comprise: (i) a cationic lipid; (ii) a phospholipid; (iii) a sterol; and (iv) a PEG-lipid conjugate. In embodiments, the lipid nanoparticles comprise: (i) a cationic lipid selected from the group consisting of DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B, DODAP, SM-102, ALC-0315, L-319, 5A2-SC8, 306Oi10, Lipid CL1, ATX-0114, MC3, DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B, DODAP, SM-102, ALC- 0315, L-319, 5A2-SC8, 306Oi10, Lipid CL1, ATX-0114, MC3, DLinDMA, DLin-KC2-DMA, DLin-KC3-DMA, DLin-KC4-DMA, DLin-K6-DMA, DLin-K-MPZ, DLin-K-DMA, DLin-C- DAP, DLin-DAC, DLin-MA, DLinDAP, DLin-S-DMA, DLin-2-DMAP, DLin-TMA, DLin- TAP, DLin-MPZ, DLinAP, DLin-EG-DMA, DLincarbDAP, and a mixture of two or more thereof; (ii) a phospholipid selected from the group consisting of DSPC, DPPC, DOPE, POPC, POPE, POPG, DPPE, DMPE, DSPE, MMPE, DMPE, DEPE, SOPE, EPC, HSPC, DPPG, and a mixture of two or more thereof; (iii) a sterol selected from the group consisting of cholesterol,cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2′-hydroxyethyl ether, cholesteryl-4′-hydroxybutyl ether, cholesteryl hemisuccinate, and a mixture of two or more thereof; and a PEG-lipid conjugate selected from the group consisting of DMG-PEG, DPPE- PEG, DPG-PEG, DSG-PEG, DSPE-PEG, and a mixture of two or more thereof; wherein the polyethylene glycol has a molecular weight from about 1,000 Daltons to about 6,000 Daltons.

[0104] In embodiments, the lipid nanoparticles comprise: (i) a cationic lipid selected from the group consisting of MC3, DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B, DODAP, SM-102, ALC-0315, L-319, 5A2- SC8, 306Oi10, Lipid CL1, ATX-0114, MC3, and a mixture of two or more thereof; (ii) a phospholipid selected from the group consisting of DSPC, DPPC, DOPE, POPC, POPE, POPG, DPPE, DMPE, DSPE, MMPE, DMPE, DEPE, SOPE, EPC, HSPC, DPPG, and a mixture of two or more thereof; (iii) a sterol selected from the group consisting of cholesterol, cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2′-hydroxyethyl ether, cholesteryl-4′- hydroxybutyl ether, cholesteryl hemisuccinate, and a mixture of two or more thereof; and a PEG-lipid conjugate selected from the group consisting of DMG-PEG, DPPE-PEG, DPG-PEG, DSG-PEG, DSPE-PEG, and a mixture of two or more thereof; wherein the polyethylene glycol has a molecular weight from about 1,000 Daltons to about 6,000 Daltons.

[0105] In embodiments, the lipid nanoparticles comprise: (i) about 30 mole% to about 70 mole% of a cationic lipid: (ii) about 5 mole% to about 20 mole% of a phospholipid; (iii) about 20 mole% to 50 mole% of a sterol; and (iv) about 0.1 mole% to about 10 mole% of a polyethylene glycol-lipid conjugate. In embodiments, the lipid nanoparticles comprise: (i) about 40 mole% to about 60 mole% of a cationic lipid; (ii) about 5 mole% to about 15 mole% of a phospholipid; (iii) about 25 mole% to about 50 mole% of a sterol; and (iv) about 0.1 mole% to about 8 mole% of a PEG-lipid conjugate. In embodiments, the lipid nanoparticles comprise: (i) about 45 mole% to about 55 mole% of a cationic lipid; (ii) about 5 mole% to about 15 mole% of a phospholipid; (iii) about 33 mole% to about 44 mole% of a sterol; and (iv) about 0.1 mole% to about 3 mole% of a PEG-lipid conjugate. In embodiments, the lipid nanoparticles comprise: (i) about 10 mole% to about 50 mole% of a cationic lipid: (ii) about 15 mole% to about 50 mole% of a phospholipid; (iii) about 30 mole% to 60 mole% of a sterol; and (iv) about 0.1 mole% to about 10 mole% of a polyethylene glycol-lipid conjugate.

[0106] In embodiments of the lipid nanoparticles described herein, the cationic lipid is any cationic lipid (or ionizable lipid) known in the art. In embodiments, the cationic lipid is DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS,CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B, DODAP, SM-102, ALC-0315, L-319, 5A2-SC8, 306Oi10, Lipid CL1, ATX-0114, or a mixture of two or more thereof. In embodiments, the cationic lipid is DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B, DODAP, SM-102, ALC-0315, L-319, 5A2-SC8, 306Oi10, Lipid CL1, ATX-0114, MC3, DLinDMA, DLin-KC2-DMA, DLin-KC3-DMA, DLin-KC4-DMA, DLin- K6-DMA, DLin-K-MPZ, DLin-K-DMA, DLin-C-DAP, DLin-DAC, DLin-MA, DLinDAP, DLin-S-DMA, DLin-2-DMAP, DLin-TMA, DLin-TAP, DLin-MPZ, DLinAP, DLin-EG-DMA, DLincarbDAP, or a mixture of two or more thereof. In embodiments, the cationic lipid is DOTAP. In embodiments, the cationic lipid is DODAC. In embodiments, the cationic lipid is DODMA. In embodiments, the cationic lipid is DSDMA. In embodiments, the cationic lipid is DOTMA. In embodiments, the cationic lipid is DDAB. In embodiments, the cationic lipid is DC-Chol. In embodiments, the cationic lipid is DMRIE. In embodiments, the cationic lipid is DOSPA. In embodiments, the cationic lipid is DOGS. In embodiments, the cationic lipid is CLinDMA. In embodiments, the cationic lipid is CpLinDMA. In embodiments, the cationic lipid is DMOBA. In embodiments, the cationic lipid is DOcarbDAP. In embodiments, the cationic lipid is DOAP. In embodiments, the cationic lipid is C12-200. In embodiments, the cationic lipid is 98N12-5. In embodiments, the cationic lipid is Lipid 5. In embodiments, the cationic lipid is Lipid A9. In embodiments, the cationic lipid is 4A3-SC8. In embodiments, the cationic lipid is 306-012B. In embodiments, the cationic lipid is DODAP. In embodiments, the cationic lipid is SM-102. In embodiments, the cationic lipid is ALC-0315. In embodiments, the cationic lipid is L-319. In embodiments, the cationic lipid is 5A2-SC8 (chemical structure described by Buschmann et al, Vaccines, 9, 65, 2021). In embodiments, the cationic lipid is 306Oi10. In embodiments, the cationic lipid is Lipid CL1. In embodiments, the cationic lipid is ATX-0114. In embodiments, the cationic lipid is MC3. In embodiments, the cationic lipid is Dlin-KC2-DMA. In embodiments, the cationic lipid is DLinDMA. In embodiments, the cationic lipid is DLin-KC3-DMA. In embodiments, the cationic lipid is DLin-KC4-DMA. In embodiments, the cationic lipid is DLin-K6-DMA. In embodiments, the cationic lipid is DLin- K-MPZ. In embodiments, the cationic lipid is DLin-K-DMA. In embodiments, the cationic lipid is DLin-C-DAP. In embodiments, the cationic lipid is DLin-DAC. In embodiments, the cationic lipid is DLin-MA. In embodiments, the cationic lipid is DLinDAP. In embodiments, the cationic lipid is DLin-S-DMA. In embodiments, the cationic lipid is DLin-2-DMAP. In embodiments, the cationic lipid is Dlin-TMA. In embodiments, the cationic lipid is Dlin-TAP. In embodiments,the cationic lipid is DLin-MPZ. In embodiments, the cationic lipid is DLinAP. In embodiments, the cationic lipid is DLin-EG-DMA. In embodiments, the cationic lipid is DLincarbDAP.

[0107] In embodiments of the lipid nanoparticles described herein, the phospholipid is any phospholipid known in the art. In embodiments, the phospholipid is DSPC, DPPC, DOPE, POPC, DOPC, POPE, POPG, DPPE, DMPE, DSPE, MMPE, DMPE, DEPE, SOPE, EPC, HSPC, DPPG. or a mixture of two or more thereof. In embodiments, the phospholipid is DSPC. In embodiments, the phospholipid is DPPC. In embodiments, the phospholipid is DOPE. In embodiments, the phospholipid is POPC. In embodiments, the phospholipid is POPE. In embodiments, the phospholipid is POPG. In embodiments, the phospholipid is DPPE. In embodiments, the phospholipid is DMPE. In embodiments, the phospholipid is DSPE. In embodiments, the phospholipid is MMPE. In embodiments, the phospholipid is DMPE. In embodiments, the phospholipid is DEPE. In embodiments, the phospholipid is SOPE. In embodiments, the phospholipid is EPC. In embodiments, the phospholipid is HSPC. In embodiments, the phospholipid is DPPG. In embodiments, the phospholipid is DOPC.

[0108] In embodiments of the lipid nanoparticles described herein, the sterol is any sterol known in the art. In embodiments, the sterol is cholesterol, cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2′-hydroxyethyl ether, cholesteryl-4′-hydroxybutyl ether, cholesteryl hemisuccinate, or a mixture of two or more thereof. In embodiments, the sterol is cholesterol. In embodiments, the sterol is cholestanol. In embodiments, the sterol is cholestanone. In embodiments, the sterol is cholestenone. In embodiments, the sterol is coprostanol. In embodiments, the sterol is cholesteryl-2′-hydroxyethyl ether. In embodiments, the sterol is cholesteryl-4′-hydroxybutyl ether. In embodiments, the sterol is cholesteryl hemisuccinate.

[0109] In embodiments of the lipid nanoparticles described herein, the PEG-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,000 Daltons to about 6,000 Daltons conjugated to a C12-C22fatty acid lipid. In embodiments, the PEG-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,000 Daltons to about 6,000 Daltons conjugated to a C12-C20fatty acid lipid. In embodiments, the PEG-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,000 Daltons to about 6,000 Daltons conjugated to a C12-C18fatty acid lipid. In embodiments, the PEG-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,000 Daltons to about 5,000 Daltons conjugated to a C12-C22fatty acid lipid. In embodiments, the PEG-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,000 Daltons to about 4,000 Daltons conjugated to a C12-C20fatty acid lipid. In embodiments, the PEG-lipid conjugate is apolyethylene glycol having a molecular weight from about 1,500 Daltons to about 4,000 Daltons conjugated to a C12-C18 fatty acid lipid. In embodiments, the PEG-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,500 Daltons to about 3,000 Daltons conjugated to a C12-C18 fatty acid lipid. In embodiments, the PEG-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,500 Daltons to about 2,500 Daltons conjugated to a C12 fatty acid lipid. In embodiments, the fatty acid lipid is saturated. In embodiments, the fatty acid lipid is unsaturated. In embodiments, the fatty acid lipid comprises one, two, or three –CH=CH- groups. In embodiments, the fatty acid lipid comprises one – CH=CH- group. In embodiments, the fatty acid lipid comprises two –CH=CH- groups. In embodiments, the PEG-lipid conjugate is DMG-PEG, DPPE-PEG, DPG-PEG, DSG-PEG, DSPE-PEG, or a mixture of two or more thereof. In embodiments, the PEG-lipid conjugate is DMG-PEG. In embodiments, the PEG-lipid conjugate is DPPE-PEG. In embodiments, the PEG- lipid conjugate is DPG-PEG. In embodiments, the PEG-lipid conjugate is DSG-PEG. In embodiments, the PEG-lipid conjugate is DSPE-PEG. In embodiments, the PEG-lipid conjugate is DMG-PEG2000, DPPE-PEG2000, DPG-PEG2000, DSG-PEG2000, DSPE-PEG2000, or a mixture of two or more thereof, wherein PEG2000 refers to PEG having a molecular weight of about 2,000 Daltons. In embodiments, the PEG-lipid conjugate is DMG-PEG2000. In embodiments, the PEG-lipid conjugate is DPPE-PEG2000. In embodiments, the PEG-lipid conjugate is DPG-PEG2000. In embodiments, the PEG-lipid conjugate is DSG-PEG2000. In embodiments, the PEG-lipid conjugate is DSPE-PEG2000.

[0110] In embodiments, the lipid nanoparticles comprise (i) Lipid 5, Lipid A9, SM-102, ALC- 0315, C12-200, 306-O12B, 4A3-SC8, DODAP, MC3, or a combination of two or more thereof; (ii) DPPC, DSPC, DOPE, DOPC, or a combination of two or more thereof; (iii) cholesterol; and (iv) DMG-PEG2000. In embodiments, the lipid nanoparticles comprise (i) Lipid 5, Lipid A9, SM-102, ALC-0315, C12-200, 306-O12B, 4A3-SC8, DODAP, or MC3; (ii) DPPC, DSPC, DOPE, or DOPC; (iii) cholesterol; and (iv) DMG-PEG2000.

[0111] In embodiments, the lipid nanoparticles comprise (i) Lipid 5, Lipid A9, SM-102, ALC- 0315, C12-200, 306-O12B, 4A3-SC8, DODAP, or a combination of two or more thereof; (ii) DPPC, DSPC, DOPE, DOPC, or a combination of two or more thereof; (iii) cholesterol; and (iv) DMG-PEG2000. In embodiments, the lipid nanoparticles comprise (i) Lipid 5, Lipid A9, SM- 102, ALC-0315, C12-200, 306-O12B, 4A3-SC8, or DODAP; (ii) DPPC, DSPC, DOPE, or DOPC; (iii) cholesterol; and (iv) DMG-PEG2000.

[0112] In embodiments, the lipid nanoparticles comprise Lipid 5, DPPC, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 40 mol% to about 60mol% of Lipid 5, about 5 mol% to about 15 mol% of DPPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 50 mol% of Lipid 5, about 10 mol% of DPPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0113] In embodiments, the lipid nanoparticles comprise Lipid A9, DSPC, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 40 mol% to about 60 mol% of Lipid A9, about 5 mol% to about 15 mol% of DSPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 50 mol% of Lipid A9, about 10 mol% of DSPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0114] In embodiments, the lipid nanoparticles comprise SM-102, DSPC, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 40 mol% to about 60 mol% of SM-102, about 5 mol% to about 15 mol% of DSPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 50 mol% of SM-102, about 10 mol% of DSPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0115] In embodiments, the lipid nanoparticles comprise ALC-0315, DSPC, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 40 mol% to about 60 mol% of ALC-0315, about 5 mol% to about 15 mol% of DSPC, about 36 mol% to about 48 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 46.3 mol% of ALC-0315, about 9.4 mol% of DSPC, about 42.7 mol% of cholesterol, and about 1.6 mol% of DMG-PEG2000.

[0116] In embodiments, the lipid nanoparticles comprise C12-200, DOPE, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 30 mol% to about 40 mol% of C12-200, about 10 mol% to about 20 mol% of DOPE, about 40 mol% to about 50 mol% of cholesterol, and about 1 mol% to about 4 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 35 mol% of C12-200, about 16 mol% of DOPE, about 46.5 mol% of cholesterol, and about 2.5 mol% of DMG-PEG2000.

[0117] In embodiments, the lipid nanoparticles comprise 306-O12B, DOPC, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 40 mol% to about 60 mol% of 306-O12B, about 5 mol% to about 15 mol% of DOPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 50 mol% of 306-O12B, about 10 mol% of DOPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0118] In embodiments, the lipid nanoparticles comprise 4A3-SC8, DOPE, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 15 mol% to about 30 mol% of 4A3-SC8, about 15 mol% to about 30 mol% of DOPE, about 40 mol% to about 55 mol% of cholesterol, and about 3 mol% to about 7 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 23.8 mol% of 4A3-SC8, about 23.8 mol% of DOPE, about 47.6 mol% of cholesterol, and about 4.8 mol% of DMG-PEG2000.

[0119] In embodiments, the lipid nanoparticles comprise 4A3-SC8, DODAP, DOPE, cholesterol, and DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 15 mol% to about 25 mol% of 4A3-SC8, about 15 mol% to about 30 mol% of DODAP, about 15 mol% to about 30 mol% of DOPE, about 30 mol% to about 50 mol% of cholesterol, and about 2 mol% to about 6 mol% of DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 19 mol% of 4A3-SC8, about 20 mol% of DODAP, about 19 mol% of DOPE, about 38 mol% of cholesterol, and about 3.8 mol% of DMG-PEG2000.

[0120] In embodiments, the lipid nanoparticles comprise MC3, DSPC, cholesterol, and DMG- PEG2000. In embodiments, the lipid nanoparticles comprise about 40 mol% to about 60 mol% of MC3, about 5 mol% to about 15 mol% DSPC, about 33 mol% to about 44 mol% cholesterol, and about 2 mol% to about 10 mol% DMG-PEG2000. In embodiments, the lipid nanoparticles comprise about 50 mol% of MC3, about 10 mol% DSPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0121] The lipid nanoparticles described herein typically have an average size (e.g., mean diameter) from about 10 nm to about 200 nm, from about 20 nm to about 190 nm, from about 30 nm to about 175 nm, from about 40 nm to about 160 nm, from about 50 nm to about 150 nm, or from about 60 nm to about 140 nm. In embodiments, the lipid nanoparticles described herein have an average size from about 30 nm to about 130 nm. In embodiments, the lipid nanoparticles have an average size from about 35 nm to about 125 nm. In embodiments, the lipid nanoparticles have an average size from about 40 nm to about 120 nm. In embodiments, the lipid nanoparticles have an average size from about 45 nm to about 115 nm. In embodiments, the lipid nanoparticles have an average size from about 50 nm to about 110 nm. In embodiments, the lipid nanoparticles have an average size from about 55 nm to about 105 nm. In embodiments, the lipid nanoparticles have an average size from about 60 nm to about 105 nm. In embodiments, the lipid nanoparticles have an average size from about 60 nm to about 100 nm. In embodiments, the lipid nanoparticles have an average size from about 65 nm to about 95 nm. In embodiments, the lipid nanoparticles have an average size from about 70 nm to about 90 nm. In embodiments, the lipid nanoparticles have an average size from about 75 nmto about 85 nm. In embodiments, the lipid nanoparticles have an average size from about 75 nm to about 80 nm. In embodiments, the lipid nanoparticles have an average size from about 70 nm to about 110 nm. In embodiments, the lipid nanoparticles have an average size from about 70 nm to about 105 nm. In embodiments, the lipid nanoparticles have an average size from about 70 nm to about 100 nm. In embodiments, the lipid nanoparticles have an average size from about 90 nm to about 110 nm. In embodiments, the lipid nanoparticles have an average size from about 95 nm to about 105 nm. In embodiments, the lipid nanoparticles have an average size from about 95 nm to about 100 nm. In embodiments, the lipid nanoparticles have an average size of about 70 nm. In embodiments, the lipid nanoparticles have an average size of about 75 nm. In embodiments, the lipid nanoparticles have an average size of about 80 nm. In embodiments, the lipid nanoparticles have an average size of about 85 nm. In embodiments, the lipid nanoparticles have an average size of about 90 nm. In embodiments, the lipid nanoparticles have an average size of about 95 nm. In embodiments, the lipid nanoparticles have an average size of about 100 nm. In embodiments, lipid nanoparticles refers to a plurality of lipid nanoparticles.

[0122] “Zeta potential” is a measure of the effective electric charge on the nanoparticle surface. The magnitude of the zeta potential provides information about particle stability, with particles with higher magnitude zeta potentials exhibiting increased stability due to a larger electrostatic repulsion between particles. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -1 mV to about -50 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -1 mV to about -10 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -1 mV to about -50 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -1 mV to about -5 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -1 mV to about -45 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -5 mV to about -40 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -5 mV to about -35 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -8 mV to about -26 mV. In embodiments, the lipid nanoparticles described herein have a zeta potential from about -10 mV to about -26 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -11 mV to about -25 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -12 mV to about -24 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -13 mV to about -23 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -14 mV to about -22 mV.In embodiments, the lipid nanoparticles have a zeta potential from about -15 mV to about -21 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -16 mV to about - 20 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -17 mV to about -20 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -17 mV to about -19 mV. In embodiments, the lipid nanoparticles have a zeta potential from about -18 mV to about -19 mV. In embodiments, the lipid nanoparticles have a zeta potential from about - 19 mV to about -20 mV. In embodiments, the lipid nanoparticles have a zeta potential of about - 12 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -13 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -14 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -15 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -16 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -17 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -18 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -19 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -20 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -21 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -22 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -23 mV. In embodiments, the lipid nanoparticles have a zeta potential of about -24 mV.

[0123] Lipid Nanoparticles Encapsulating Nucleic Acids

[0124] In embodiments, the disclosure provides a lipid nanoparticle comprising a nucleic acid encapsulated within the lipid nanoparticle. For purposes of the disclosure, the phrase “lipid nanoparticle comprises a nucleic acid” is equivalent to the phrase “lipid nanoparticle comprises a nucleic acid encapsulated within the lipid nanoparticle.” In embodiments, the nucleic acid is any nucleic acid described herein, including embodiments thereof. In embodiments, the nucleic acid encapsulated within the lipid nanoparticle is RNA. In embodiments, the nucleic acid encapsulated within the lipid nanoparticle is mRNA.

[0125] In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ALDH2 enzyme. In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:7, SEQ ID NO:11, or SEQ ID NO:14 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:7 (or a percent sequence identity thereof asdescribed herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:11 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:14 (or a percent sequence identity thereof as described herein).

[0126] In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:21 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:1 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:3 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:9 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:12 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ALDH2 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:21 (or a percent sequence identitythereof as described herein).

[0127] In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ADH1 enzyme. In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:8, SEQ ID NO:17, or SEQ ID NO:20 (or a percent sequence identity thereof as described herein). In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:17 (or a percent sequence identity thereof as described herein). In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:20 (or a percent sequence identity thereof as described herein).

[0128] In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid sequence comprises SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24 (or a percentsequence identity thereof as described herein). In embodiments, the lipid nanoparticle asdescribed herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:4 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:6 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:15 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:16 (or a percentsequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:18 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid encoding an ADH1 enzyme, wherein the nucleic acid comprises SEQ ID NO:24 (or a percent sequence identity thereof as described herein).

[0129] In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ALDH2 enzyme and an ADH1 enzyme. In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: (i) an ALDH2 enzyme having SEQ ID NO:7, SEQ ID NO:11, or SEQ ID NO:14 (or a percent sequence identity thereof as described herein) and (ii) an ADH1 enzyme having SEQ ID NO:8, SEQ ID NO:17, or SEQ ID NO:20 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:7 (or a percent sequence identity thereof as described herein) and SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:7 (or a percent sequence identity thereof as described herein) and SEQ ID NO:17 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:7 (or a percent sequence identity thereof as described herein) and SEQ ID NO:20 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:11 (or a percent sequence identity thereof as described herein) and SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments,the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:11 (or a percent sequence identity thereof as described herein) and SEQ ID NO:17 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:11 (or a percent sequence identity thereof as described herein) and SEQ ID NO:20 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:14 (or a percent sequence identity thereof as described herein) and SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:14 (or a percent sequence identity thereof as described herein) and SEQ ID NO:17 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid that encodes: SEQ ID NO:14 (or a percent sequence identity thereof as described herein) and SEQ ID NO:20 (or a percent sequence identity thereof as described herein).

[0130] In embodiments, the disclosure provides a lipid nanoparticle as described herein (including any embodiment thereof) comprising a nucleic acid encoding an ALDH2 enzyme and an ADH1 enzyme. In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises: (i) SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and (ii) SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:10(or a percent sequence identity thereof as described herein) and SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof)comprises a nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the lipid nanoparticle as described herein (including any embodiment thereof) comprises a nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and SEQ ID NO:24 (or a percent sequence identity thereof as described herein).

[0131] In embodiments, the disclosure provides a plurality of lipid nanoparticles as described herein (including any embodiment thereof) comprising a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme and a second lipid nanoparticle whichcomprises a second nucleic acid encoding an ADH1 enzyme. In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acidwhich comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:5 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:1 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:4 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:3 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:6 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identitythereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:22 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:23 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:2 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:24 (or a percent sequence identity thereof as described herein). In embodiments, the first lipid nanoparticle comprises a first nucleic acid which comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and the second lipid nanoparticle comprises a second nucleic acid which comprises SEQ ID NO:24 (or a percent sequence identity thereof as described herein).

[0132] In embodiments, the disclosure provides a plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprising a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme and a second lipid nanoparticle which comprises a second nucleic acid encoding a ADH1 enzyme. In embodiments, the plurality oflipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:7, SEQ ID NO:11, or SEQ ID NO:14 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:8, SEQ ID NO:17, or SEQ ID NO:20 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:7 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:7 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:7 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein theADH1 enzyme comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:10 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:13 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:8 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle whichcomprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:16 (or a percent sequence identity thereof as described herein). In embodiments, the plurality of lipid nanoparticle as described herein (including any embodiment thereof) comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme, wherein the ALDH2 enzyme comprises SEQ ID NO:21 (or a percent sequence identity thereof as described herein) and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, wherein the ADH1 enzyme comprises SEQ ID NO:19 (or a percent sequence identity thereof as described herein).

[0133] In embodiments, where the plurality of lipid nanoparticles comprises a first lipid nanoparticle which comprises a first nucleic acid encoding an ALDH2 enzyme and a second lipid nanoparticle which comprises a second nucleic acid encoding an ADH1 enzyme, any amount or ratio of the first lipid nanoparticle and the second nanoparticle can be present in the plurality of lipid nanoparticles. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 99:1 to about 1:99 (e.g., of the total of the plurality of lipid nanoparticles, about 99% are a plurality of the first lipid nanoparticle and about 1% are a plurality of the second lipid nanoparticle). In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 95:5 to about 5:95. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 90:10 to about 10:90. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 85:15 to about 15:85. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 80:20 to about 20:80. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 75:25 to about 25:75. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 70:30 to about 30:70. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 65:35 to about 35:65. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 60:40 to about 40:60. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is from about 55:45 to about 45:55. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 50:50. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 95:5. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 90:10. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 85:15. In embodiments, the percent ratioof the first lipid nanoparticles to the second lipid nanoparticles is about 80:20. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 75:25. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 70:30. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 65:35. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 60:40. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 55:45. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 5:95. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 10:90. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 15:85. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 20:80. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 25:75. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 30:70. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 35:65. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 40:60. In embodiments, the percent ratio of the first lipid nanoparticles to the second lipid nanoparticles is about 45:55.

[0134] The term “N / P” or “N / P ratio” refers to the ratio of positively-chargeable polymer amine (N = nitrogen) groups to negatively-charged nucleic acid phosphate (P) groups in a lipid encapsulated nanoparticle. In embodiments of the lipid nanoparticles having a nucleic acid encapsulated therein, the N / P ratio is from about 1:1 to about 15:1. In embodiments, the N / P ratio is from about 1:1 to about 10:1. In embodiments, the N / P ratio is from about 2:1 to about 8:1. In embodiments, the N / P ratio is from about 2:1 to about 7:1. In embodiments, the N / P ratio is from about 3:1 to about 6:1. In embodiments, the N / P ratio is from about 2:1 to about 4:1. In embodiments, the N / P ratio is from about 5:1 to about 7:1. In embodiments, the N / P ratio is about 1:1. In embodiments, the N / P ratio is about 2:1. In embodiments, the N / P ratio is about 3:1. In embodiments, the N / P ratio is about 4:1. In embodiments, the N / P ratio is about 5:1. In embodiments, the N / P ratio is about 6:1. In embodiments, the N / P ratio is about 7:1. In embodiments, the N / P ratio is about 8:1. In embodiments, the N / P ratio is about 9:1. In embodiments, the N / P ratio is about 10:1.

[0135] Pharmaceutical Compositions

[0136] Provided herein are pharmaceutical compositions comprising a lipid nanoparticle which comprises a nucleic acid encapsulated therein and a pharmaceutically acceptableexcipient. Provided herein are pharmaceutical compositions comprising a plurality of lipid nanoparticles which comprise nucleic acids encapsulated therein and a pharmaceutically acceptable excipient.

[0137] “Pharmaceutically acceptable excipient” and “pharmaceutically acceptable carrier” refer to a substance that aids the administration of an active agent to and absorption by a subject and can be included in the compositions of the disclosure without causing a significant adverse toxicological effect on the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions, alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure. One of skill in the art will recognize that other pharmaceutical excipients are useful.

[0138] Solutions of the nucleic acids or lipid nanoparticles containing nucleic acids can be prepared in water suitably mixed with a lipid or surfactant, such as hydroxypropyl cellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations can contain a preservative to prevent the growth of microorganisms.

[0139] Pharmaceutical compositions can be delivered via intranasal or inhalable solutions. The intranasal composition can be a spray, aerosol, or inhalant. The inhalable composition can be a spray, aerosol, or inhalant. Nasal solutions can be aqueous solutions designed to be administered to the nasal passages in drops or sprays. Nasal solutions can be prepared so that they are similar in many respects to nasal secretions. Thus, the aqueous nasal solutions usually are isotonic and slightly buffered to maintain a pH of 5.5 to 6.5. In addition, antimicrobial preservatives, similar to those used in ophthalmic preparations and appropriate drug stabilizers, if required, may be included in the formulation. Various commercial nasal preparations are known in the art.

[0140] Oral formulations can include excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders. In embodiments, oral pharmaceutical compositions will comprise an inert diluent or edible carrier, or they may be enclosed in hard orsoft shell gelatin capsule, or they may be compressed into tablets, or they may be incorporated directly with the food. For oral therapeutic administration, the active compounds may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. The percentage of the compositions and preparations may, of course, be varied and may be between about 1 to about 75% of the weight of the unit. The amount of nucleic acids in such compositions is such that a suitable dosage can be obtained.

[0141] For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered and the liquid diluent first rendered isotonic with sufficient saline or glucose. Aqueous solutions, in particular, sterile aqueous media, are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. For example, one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or injected at the proposed site of infusion.

[0142] Sterile injectable solutions can be prepared by incorporating the nucleic acids in the required amount in the appropriate solvent followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium. Vacuum-drying and freeze-drying techniques, which yield a powder of the active ingredient plus any additional desired ingredients, can be used to prepare sterile powders for reconstitution of sterile injectable solutions. The preparation of more, or highly, concentrated solutions for direct injection is also contemplated. Dimethyl sulfoxide can be used as solvent for extremely rapid penetration, delivering high concentrations of the active agents to a small area.

[0143] The formulations of nucleic acids, lipid nanoparticles containing nucleic acids can be presented in unit-dose or multi-dose sealed containers, such as nebulizers, ventilators, ampules, and vials. Thus, the composition can be in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of nucleic acids or lipid nanoparticles containing nucleic acids. Thus, the compositions can be administered in a variety of unit dosage forms depending upon the method of administration. For example, unit dosage forms suitable for oral administration include, but are not limited to, powder, tablets, pills, capsules and lozenges.

[0144] The nucleic acids, lipid nanoparticles containing nucleic acids, and pharmaceutical compositions can be administered to the patient in any manner as described herein. In embodiments, the nucleic acids, lipid nanoparticles, and pharmaceutical compositions are administered parenterally to a patient. In embodiments, the nucleic acids, lipid nanoparticles,and pharmaceutical compositions are administered intravenously to a patient. In embodiments, the nucleic acids, lipid nanoparticles, and pharmaceutical compositions are administered subcutaneously to a patient. In embodiments, the nucleic acids, lipid nanoparticles, and pharmaceutical compositions are administered intranodally to a patient. In embodiments, the nucleic acids, lipid nanoparticles, and pharmaceutical compositions are administered intratumorally to a patient.

[0145] Methods

[0146] Provided herein are methods of treating or preventing alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the nucleic acids, enzymes, lipid nanoparticles, and pharmaceutical compositions described herein, including embodiments thereof. In embodiments, the disclosure provides methods of treating or preventing alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the lipid nanoparticles described herein, including embodiments thereof, wherein the lipid nanoparticles comprise the nucleic acids described herein, including embodiments thereof. In embodiments, the disclosure provides methods of treating or preventing alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the pharmaceutical compositions described herein, including embodiments thereof.

[0147] Provided herein are methods of treating alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the nucleic acids, enzymes, lipid nanoparticles, and pharmaceutical compositions described herein, including embodiments thereof. In embodiments, the disclosure provides methods of treating alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the lipid nanoparticles described herein, including embodiments thereof, wherein the lipid nanoparticles comprise the nucleic acids described herein, including embodiments thereof. In embodiments, the disclosure provides methods of treating alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the pharmaceutical compositions described herein, including embodiments thereof.

[0148] Provided herein are methods of preventing alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the nucleic acids, enzymes, lipid nanoparticles, and pharmaceutical compositions described herein, including embodiments thereof. In embodiments, the disclosure provides methods of preventing alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the lipid nanoparticles described herein, including embodiments thereof, wherein thelipid nanoparticles comprise the nucleic acids described herein, including embodiments thereof. In embodiments, the disclosure provides methods of preventing alcohol poisoning in a patient in need thereof comprising administering to the patient an effective amount of the pharmaceutical compositions described herein, including embodiments thereof.

[0149] In embodiments, the method of treating or preventing alcohol poisoning is a method of treating or preventing a symptom resulting from alcohol poisoning, such as mental confusion, hypoglycemia, seizure, slowed breathing, irregular breathing, hypothermia, irregular heartbeat, slow heart rate, loss of consciousness, emesis, heart attack, coma, dehydration, or a combination of two or more thereof. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing mental confusion. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing hypoglycemia. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing slowed breathing or irregular breathing. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing hypothermia. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing irregular heartbeat or slow heart rate. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing loss of consciousness. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing emesis. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing heart attack. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing coma. In embodiments, the method of treating alcohol poisoning is a method of treating or preventing dehydration.

[0150] Alcohol poisoning can alternatively be referred to as alcohol overdose. In embodiments, alcohol poisoning is marked by a blood alcohol level of about 0.05% or higher. In embodiments, alcohol poisoning is marked by a blood alcohol level of about 0.15% or higher. In embodiments, alcohol poisoning is marked by a blood alcohol level of about 0.30% or higher. In embodiments, alcohol poisoning is marked by a blood alcohol level of about 0.45% or higher. In embodiments, alcohol poisoning is marked by a blood alcohol level of about 0.60% or higher.

[0151] Provided herein are methods of treating or preventing alcohol flush syndrome in a patient in need thereof comprising administering to the patient an effective amount of the nucleic acids, enzymes, and lipid nanoparticles described herein, including embodiments thereof. In embodiments, the disclosure provides methods of treating or preventing alcohol flush syndrome in a patient in need thereof comprising administering to the patient an effective amount of the lipid nanoparticles described herein, including embodiments thereof, wherein the lipid nanoparticles comprise the nucleic acids described herein, including embodiments thereof. Inembodiments, the disclosure provides methods of treating or preventing alcohol flush syndrome in a patient in need thereof comprising administering to the patient an effective amount of the pharmaceutical compositions described herein, including embodiments thereof. In embodiments, the patient is East Asian (e.g., Korean, Chinese, Japanese) or of East Asian-descent. In embodiments, the patient has a mutant ALDH2*2 allele that renders the ALDH2 enzyme inactive. In embodiments, the patient is homozygous for ALDH2 (ALDH2*2 / *2). In embodiments, the patient is heterozygous for ALDH2 (ALDH2*1 / *2).

[0152] In embodiments, the method of treating or preventing alcohol poisoning further comprises administering to the patient an effective amount of an enzymatic cofactor, supplemental magnesium, supplemental zinc, or a combination of two or more thereof.

[0153] In embodiments, the method of treating or preventing a symptom resulting from alcohol poisoning further comprises administering to the patient an effective amount of an enzymatic cofactor, supplemental magnesium, supplemental zinc, or a combination of two or more thereof.

[0154] In embodiments, the method of treating or preventing alcohol flush syndrome further comprises administering to the patient an effective amount of an enzymatic cofactor, supplemental magnesium, supplemental zinc, or a combination of two or more thereof.

[0155] In embodiments, the method of treating or preventing alcohol poisoning further comprises administering to the patient an effective amount of oxidized nicotinamide adenine dinucleotide (NAD+), reduced nicotinamide adenine dinucleotide (NADH), nicotinamide adenine dinucleotide, niacin, nicotinamide, nicotinamide riboside, dihydronicotinamide riboside, nicotinamide mononucleotide, dihydronicotinamide mononucleotide, tryptophan, or a combination of two or more thereof.

[0156] In embodiments, the method of treating or preventing a symptom resulting from alcohol poisoning further comprises administering to the patient an effective amount of oxidized nicotinamide adenine dinucleotide (NAD+), reduced nicotinamide adenine dinucleotide (NADH), nicotinamide adenine dinucleotide, niacin, nicotinamide, nicotinamide riboside, dihydronicotinamide riboside, nicotinamide mononucleotide, dihydronicotinamide mononucleotide, tryptophan, or a combination of two or more thereof. In embodiments, the method further comprises administering supplemental magnesium, supplemental zinc, or a combination thereof.

[0157] In embodiments, the method of treating or preventing alcohol flush syndrome further comprises administering to the patient an effective amount of oxidized nicotinamide adenine dinucleotide (NAD+), reduced nicotinamide adenine dinucleotide (NADH), nicotinamideadenine dinucleotide, niacin, nicotinamide, nicotinamide riboside, dihydronicotinamide riboside, nicotinamide mononucleotide, dihydronicotinamide mononucleotide, tryptophan, or a combination of two or more thereof. In embodiments, the method further comprises administering supplemental magnesium, supplemental zinc, or a combination thereof.

[0158] In embodiments, the enzymatic cofactor is oxidized nicotinamide adenine dinucleotide (NAD+). In embodiments, the enzymatic cofactor is a precursor of oxidized nicotinamide adenine dinucleotide (NAD+). In embodiments, the enzymatic cofactor is oxidized nicotinamide adenine dinucleotide (NAD+), a precursor of oxidized nicotinamide adenine dinucleotide (NAD+), or a combination thereof. In embodiments, the precursor of NAD+ is nicotinamide adenine dinucleotide, niacin, nicotinamide, nicotinamide riboside, dihydronicotinamide riboside, nicotinamide mononucleotide, dihydronicotinamide mononucleotide, tryptophan, or a combination of two or more thereof. In embodiments, the precursor of NAD+ is niacin, nicotinamide, nicotinamide riboside, nicotinamide mononucleotide, tryptophan, or a combination of two or more thereof. In embodiments, the precursor of NAD+ is niacin. In embodiments, the precursor of NAD+ is nicotinamide. In embodiments, the precursor of NAD+ is nicotinamide riboside. In embodiments, the precursor of NAD+ is nicotinamide mononucleotide. In embodiments, the precursor of NAD+ is tryptophan. In embodiments, the effective amount of the enzymatic cofactor is an amount that produces a plasma concentration of up to about 20 mM of NAD+. In embodiments, the effective amount of the enzymatic cofactor is an amount that produces a plasma concentration of up to about 15 mM of NAD+. In embodiments, the effective amount of the enzymatic cofactor is an amount that produces a plasma concentration of up to about 10 mM of NAD+. In embodiments, the effective amount of the enzymatic cofactor is an amount that produces a plasma concentration from about 2 mM to about 15 mM of NAD+. In embodiments, the effective amount of the enzymatic cofactor is an amount that produces a plasma concentration from about 2 mM to about 10 mM of NAD+.

[0159] Dose and Dosing Regimens

[0160] The dosage and frequency (single or multiple doses) of the nucleic acids, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein administered to a subject can vary depending upon a variety of factors, for example, whether the mammal suffers from another disease, and its route of administration; size, age, sex, health, body weight, body mass index, and diet of the recipient; nature and extent of symptoms of the disease being treated, kind of concurrent treatment, complications from the disease being treated or other health-related problems. Other therapeutic regimens or agents can be used in conjunction with the methods andnucleic acids, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein described herein. Adjustment and manipulation of established dosages (e.g., frequency and duration) are within the ability of the skilled artisan.

[0161] For any nucleic acid, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein, as described herein, the effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of nucleic acids, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art. As is known in the art, effective amounts of nucleic acids, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring effectiveness and adjusting the dosage upwards or downwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods is well within the capabilities of the ordinarily skilled artisan.

[0162] Dosages of the nucleic acids, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein may be varied depending upon the requirements of the patient. The dose administered to a patient should be sufficient to affect a beneficial therapeutic response in the patient over time. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects. Determination of the proper dosage for a particular situation is within the skill of the art. Dosage amounts and intervals can be adjusted individually to provide levels of the nucleic acids, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein effective for the particular clinical indication being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual's disease state.

[0163] Utilizing the teachings provided herein, an effective prophylactic or therapeutic treatment regimen can be planned that does not cause substantial toxicity and yet is effective to treat the clinical disease or symptoms demonstrated by the particular patient. This planning should involve the careful choice of nucleic acids, lipid nanoparticles having nucleic acidsencapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein by considering factors such as compound potency, relative bioavailability, patient body weight, presence and severity of adverse side effects.

[0164] In embodiments, the nucleic acid, lipid nanoparticles having nucleic acids encapsulated therein, or pharmaceutical compositions lipid nanoparticles having nucleic acids encapsulated therein is administered to a patient at an amount of about 0.001 mg / kg to about 500 mg / kg. In embodiments, the nucleic acids is administered to a patient in an amount of about 0.01 mg / kg, 0.1 mg / kg, 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 200 mg / kg, or 300 mg / kg. It is understood that where the amount is referred to as “mg / kg,” the amount is milligram per kilogram body weight of the subject being administered with the nucleic acid. In embodiments, the nucleic acid is administered to a patient in an amount from about 0.001 mg to about 500 mg per day, as a single dose, or in a dose administered two or three times per day.

[0165] Embodiments 1-265

[0166] Embodiment 1. A lipid nanoparticle comprising a nucleic acid encoding an aldehyde dehydrogenase 2 enzyme.

[0167] Embodiment 2. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:1.

[0168] Embodiment 3. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:1.

[0169] Embodiment 4. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:2

[0170] Embodiment 5. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:2.

[0171] Embodiment 6. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:3.

[0172] Embodiment 7. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:3.

[0173] Embodiment 8. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:9.

[0174] Embodiment 9. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:9.

[0175] Embodiment 10. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:10.

[0176] Embodiment 11. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:10.

[0177] Embodiment 12. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:12.

[0178] Embodiment 13. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:12.

[0179] Embodiment 14. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:13.

[0180] Embodiment 15. The lipid nanoparticle of embodiment 1, wherein the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:13.

[0181] Embodiment 16. The lipid nanoparticle of embodiment 1, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:7.

[0182] Embodiment 17. The lipid nanoparticle of embodiment 1, wherein the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:7.

[0183] Embodiment 18. The lipid nanoparticle of embodiment 1, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:11.

[0184] Embodiment 19. The lipid nanoparticle of embodiment 1, wherein the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:11.

[0185] Embodiment 20. The lipid nanoparticle of embodiment 1, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:14.

[0186] Embodiment 21. The lipid nanoparticle of embodiment 1, wherein the aldehyde dehydrogenase 2 enzyme has SEQ ID NO:14.

[0187] Embodiment 22. A lipid nanoparticle comprising a nucleic acid encoding an alcohol dehydrogenase 1 enzyme.

[0188] Embodiment 23. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:4.

[0189] Embodiment 24. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has SEQ ID NO:4.

[0190] Embodiment 25. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:5.

[0191] Embodiment 26. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has SEQ ID NO:5.

[0192] Embodiment 27. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:6.

[0193] Embodiment 28. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has SEQ ID NO:6.

[0194] Embodiment 29. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:15.

[0195] Embodiment 30. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has SEQ ID NO:15.

[0196] Embodiment 31. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:16.

[0197] Embodiment 32. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has SEQ ID NO:16.

[0198] Embodiment 33. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:18.

[0199] Embodiment 34. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has SEQ ID NO:18.

[0200] Embodiment 35. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:19.

[0201] Embodiment 36. The lipid nanoparticle of embodiment 22, wherein the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has SEQ ID NO:19.

[0202] Embodiment 37. The lipid nanoparticle of embodiment 22, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:8.

[0203] Embodiment 38. The lipid nanoparticle of embodiment 22, wherein the alcohol dehydrogenase 1 enzyme has SEQ ID NO:8.

[0204] Embodiment 39. The lipid nanoparticle of embodiment 22, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:17.

[0205] Embodiment 40. The lipid nanoparticle of embodiment 22, wherein the alcohol dehydrogenase 1 enzyme has SEQ ID NO:17.

[0206] Embodiment 41. The lipid nanoparticle of embodiment 22, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:20.

[0207] Embodiment 42. The lipid nanoparticle of embodiment 22, wherein the alcohol dehydrogenase 1 enzyme has SEQ ID NO:20.

[0208] Embodiment 43. A lipid nanoparticle comprising a nucleic acid encoding an aldehyde dehydrogenase 2 enzyme and an alcohol dehydrogenase 1 enzyme.

[0209] Embodiment 44. The lipid nanoparticle of any one of embodiment 43, wherein the nucleic acid has at least 95% sequence identity to a nucleic acid comprising: (a) SEQ ID NO:2, SEQ ID NO:10, or SEQ ID NO:13, and (b) SEQ ID NO:5, SEQ ID NO:16, or SEQ ID NO:19.

[0210] Embodiment 45. The lipid nanoparticle of any one of embodiment 43, wherein the nucleic acid sequence comprises: (a) SEQ ID NO:2, SEQ ID NO:10, or SEQ ID NO:13, and (b) SEQ ID NO:5, SEQ ID NO:16, or SEQ ID NO:19.

[0211] Embodiment 46. The lipid nanoparticle of any one of embodiment 43, wherein the nucleic acid has at least 95% sequence identity to a nucleic acid comprising: (a) SEQ ID NO:10 or SEQ ID NO:13, and (b) SEQ ID NO:16 or SEQ ID NO:19.

[0212] Embodiment 47. The lipid nanoparticle of any one of embodiment 43, wherein the nucleic acid sequence comprises: (a) SEQ ID NO:10 or SEQ ID NO:13, and (b) SEQ ID NO:16 or SEQ ID NO:19.

[0213] Embodiment 48. The lipid nanoparticle of embodiment 43, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:7.

[0214] Embodiment 49. The lipid nanoparticle of embodiment 43, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:7.

[0215] Embodiment 50. The lipid nanoparticle of embodiment 43, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:11.

[0216] Embodiment 51. The lipid nanoparticle of embodiment 43, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:11.

[0217] Embodiment 52. The lipid nanoparticle of embodiment 43, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:14.

[0218] Embodiment 53. The lipid nanoparticle of embodiment 43, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:14.

[0219] Embodiment 54. The lipid nanoparticle of any one of embodiments 43 and 48–53, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:8.

[0220] Embodiment 55. The lipid nanoparticle of any one of embodiments 43 and 48–53, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:8.

[0221] Embodiment 56. The lipid nanoparticle of any one of embodiments 43 and 48–53, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:17.

[0222] Embodiment 57. The lipid nanoparticle of any one of embodiments 43 and 48–53, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:17.

[0223] Embodiment 58. The lipid nanoparticle of any one of embodiments 43 and 48–53, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:20.

[0224] Embodiment 59. The lipid nanoparticle of any one of embodiments 43 and 48–53, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:20.

[0225] Embodiment 60. A lipid nanoparticle comprising a first nucleic acid encoding an aldehyde dehydrogenase 2 enzyme and a second nucleic acid encoding an alcohol dehydrogenase 1 enzyme.

[0226] Embodiment 61. The lipid nanoparticle of embodiment 60, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, or SEQ ID NO:13.

[0227] Embodiment 62. The lipid nanoparticle of embodiment 60, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, or SEQ ID NO:13.

[0228] Embodiment 63. The lipid nanoparticle of embodiment 60, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0229] Embodiment 64. The lipid nanoparticle of embodiment 60, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:9.

[0230] Embodiment 65. The lipid nanoparticle of embodiment 60, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:10.

[0231] Embodiment 66. The lipid nanoparticle of embodiment 60, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:12.

[0232] Embodiment 67. The lipid nanoparticle of embodiment 60, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:13.

[0233] Embodiment 68. The lipid nanoparticle of any one of embodiments 60 to 67, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, or SEQ ID NO:19.

[0234] Embodiment 69. The lipid nanoparticle of any one of embodiments 60 to 67, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, or SEQ ID NO:19.

[0235] Embodiment 70. The lipid nanoparticle of any one of embodiments 60 to 67, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6.

[0236] Embodiment 71. The lipid nanoparticle of any one of embodiments 60 to 67, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:15.

[0237] Embodiment 72. The lipid nanoparticle of any one of embodiments 60 to 67, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:16.

[0238] Embodiment 73. The lipid nanoparticle of any one of embodiments 60 to 67, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:18.

[0239] Embodiment 74. The lipid nanoparticle of any one of embodiments 60 to 67, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:19.

[0240] Embodiment 75. The lipid nanoparticle of embodiment 60, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:7, SEQ ID NO:11, or SEQ ID NO:14.

[0241] Embodiment 76. The lipid nanoparticle of embodiment 60, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:7.

[0242] Embodiment 77. The lipid nanoparticle of embodiment 60, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:11.

[0243] Embodiment 78. The lipid nanoparticle of embodiment 60, wherein the aldehydedehydrogenase 2 enzyme comprises SEQ ID NO:14.

[0244] Embodiment 79. The lipid nanoparticle of any one of embodiments 60 and 75–78, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:8, SEQ ID NO:17, or SEQ ID NO:20.

[0245] Embodiment 80. The lipid nanoparticle of any one of embodiments 60 and 75–78, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:8.

[0246] Embodiment 81. The lipid nanoparticle of any one of embodiments 60 and 75–78, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:17.

[0247] Embodiment 82. The lipid nanoparticle of any one of embodiments 60 and 75–78, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:20.

[0248] Embodiment 83. The lipid nanoparticle of any one of embodiments 1 to 82, wherein the nucleic acid is RNA.

[0249] Embodiment 84. The lipid nanoparticle of embodiment 83, wherein the RNA is mRNA.

[0250] Embodiment 85. A plurality of lipid nanoparticles comprising the lipid nanoparticle of any one of embodiments 1 to 84.

[0251] Embodiment 86. A plurality of lipid nanoparticles comprising: (i) a first lipid nanoparticle comprising a first nucleic acid encoding an aldehyde dehydrogenase 2 enzyme; and (ii) a second lipid nanoparticle comprising a second nucleic acid encoding an alcohol dehydrogenase 1 enzyme.

[0252] Embodiment 87. The lipid nanoparticles of embodiment 86, wherein the ratio of the first lipid nanoparticles to the second lipid nanoparticles is from 95:5 to 5:95.

[0253] Embodiment 88. The lipid nanoparticles of embodiment 86, wherein the ratio of the first lipid nanoparticles to the second lipid nanoparticles is from 90:10 to 10:90.

[0254] Embodiment 89. The lipid nanoparticles of embodiment 86, wherein the ratio of the first lipid nanoparticles to the second lipid nanoparticles is from 80:20 to 20:80.

[0255] Embodiment 90. The lipid nanoparticles of embodiment 86, wherein the ratio of the first lipid nanoparticles to the second lipid nanoparticles is from 70:30 to 30:70.

[0256] Embodiment 91. The lipid nanoparticles of embodiment 86, wherein the ratio of the first lipid nanoparticles to the second lipid nanoparticles is from 60:40 to 40:60.

[0257] Embodiment 92. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:1.

[0258] Embodiment 93. The lipid nanoparticles of any one of embodiments 86 to 91, whereinthe first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:1.

[0259] Embodiment 94. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:2.

[0260] Embodiment 95. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:2.

[0261] Embodiment 96. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:3.

[0262] Embodiment 97. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:3.

[0263] Embodiment 98. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:9.

[0264] Embodiment 99. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:9.

[0265] Embodiment 100. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:10.

[0266] Embodiment 101. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:10.

[0267] Embodiment 102. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:12.

[0268] Embodiment 103. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:12.

[0269] Embodiment 104. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:13.

[0270] Embodiment 105. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the first nucleic acid encoding the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:13.

[0271] Embodiment 106. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:4.

[0272] Embodiment 107. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:4.

[0273] Embodiment 108. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:5.

[0274] Embodiment 109. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:5.

[0275] Embodiment 110. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:6.

[0276] Embodiment 111. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:6.

[0277] Embodiment 112. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:15.

[0278] Embodiment 113. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:15.

[0279] Embodiment 114. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:16.

[0280] Embodiment 115. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:16.

[0281] Embodiment 116. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:18.

[0282] Embodiment 117. The lipid nanoparticles of any one of embodiments 86 to 105,wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:18.

[0283] Embodiment 118. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:19.

[0284] Embodiment 119. The lipid nanoparticles of any one of embodiments 86 to 105, wherein the second nucleic acid encoding the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:19.

[0285] Embodiment 120. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:7.

[0286] Embodiment 121. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:7.

[0287] Embodiment 122. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:11.

[0288] Embodiment 123. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:11.

[0289] Embodiment 124. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the aldehyde dehydrogenase 2 enzyme has at least 95% sequence identity to SEQ ID NO:14.

[0290] Embodiment 125. The lipid nanoparticles of any one of embodiments 86 to 91, wherein the aldehyde dehydrogenase 2 enzyme comprises SEQ ID NO:14.

[0291] Embodiment 126. The lipid nanoparticles of any one of embodiments 86 to 91 and 120 to 125, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:8.

[0292] Embodiment 127. The lipid nanoparticles of any one of embodiments 86 to 91 and 120 to 125, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:8.

[0293] Embodiment 128. The lipid nanoparticles of any one of embodiments 86 to 91 and 120 to 125, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:17.

[0294] Embodiment 129. The lipid nanoparticles of any one of embodiments 86 to 91 and 120 to 125, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:17.

[0295] Embodiment 130. The lipid nanoparticles of any one of embodiments 86 to 91 and 120to 125, wherein the alcohol dehydrogenase 1 enzyme has at least 95% sequence identity to SEQ ID NO:20.

[0296] Embodiment 131. The lipid nanoparticles of any one of embodiments 86 to 91 and 120 to 125, wherein the alcohol dehydrogenase 1 enzyme comprises SEQ ID NO:20.

[0297] Embodiment 132. The lipid nanoparticles of any one of embodiments 85 to 131, wherein the nucleic acid is RNA.

[0298] Embodiment 133. The lipid nanoparticles of embodiment 132, wherein the RNA is mRNA.

[0299] Embodiment 134. The lipid nanoparticle of any one of embodiments 1 to 84 or the plurality of lipid nanoparticles of any one of embodiments 85 to 133, wherein the lipid nanoparticle comprises: (i) a cationic lipid; (ii) a phospholipid; (iii) a sterol; and (iv) a polyethylene glycol-lipid conjugate.

[0300] Embodiment 135. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 134, comprising: (i) about 30 mole% to about 70 mole% of the cationic lipid; (ii) about 5 mole% to about 20 mole% of the phospholipid; (iii) about 20 mole% to 50 mole% of the sterol; and (iv) about 0.1 mole% to about 10 mole% of the polyethylene glycol-lipid conjugate.

[0301] Embodiment 136. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 134, comprising: (i) about 10 mole% to about 40 mole% of the cationic lipid; (ii) about 15 mole% to about 50 mole% of the phospholipid; (iii) about 30 mole% to 60 mole% of the sterol; and (iv) about 0.1 mole% to about 10 mole% of the polyethylene glycol-lipid conjugate.

[0302] Embodiment 137. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B, DODAP, SM-102, ALC-0315, L-319, 5A2-SC8, 306Oi10, Lipid CL1, ATX-0114, MC3, an MC3 derivative, DLinDMA, DLin-K-C2-DMA, DLin-K-C3-DMA, DLin-K-C4-DMA, DLin- K6-DMA, DLin-K-MPZ, DLin-K-DMA, DLin-C-DAP, DLin-DAC, DLin-MA, DLinDAP, DLin-S-DMA, DLin-2-DMAP, DLin-TMA, DLin-TAP, DLin-MPZ, DLinAP, DLin-EG-DMA, DLincarbDAP, or a mixture of two or more thereof.

[0303] Embodiment 138. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B,DODAP, SM-102, ALC-0315, L-319, 5A2-SC8, 306Oi10, Lipid CL1, ATX-0114, or a mixture of two or more thereof.

[0304] Embodiment 139. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is Lipid 5, Lipid A9, C12-200, 4A3- SC8, DODAP, 306-012B, SM-102, ALC-0315, or a mixture of two or more thereof.

[0305] Embodiment 140. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is MC3.

[0306] Embodiment 141. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is Lipid 5.

[0307] Embodiment 142. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is Lipid A9.

[0308] Embodiment 143. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is C12-200.

[0309] Embodiment 144. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is 4A3-SC8.

[0310] Embodiment 145. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is DODAP.

[0311] Embodiment 146. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid comprises 4A3-SC8 and DODAP.

[0312] Embodiment 147. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is 306-012B.

[0313] Embodiment 148. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is SM-102.

[0314] Embodiment 149. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, wherein the cationic lipid is ALC-0315.

[0315] Embodiment 150. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 149, wherein the phospholipid is DSPC, DPPC, DOPE, POPC, POPE, POPG, DPPE, DMPE, DSPE, MMPE, DEPE, SOPE, EPC, HSPC, DPPG, or a mixture of two or more thereof.

[0316] Embodiment 151. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 149, wherein the phospholipid is DSPC, DPPC, DOPE, DOPC, or a mixture of two or more thereof.

[0317] Embodiment 152. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 149, wherein the phospholipid is DSPC.

[0318] Embodiment 153. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 149, wherein the phospholipid is DPPC.

[0319] Embodiment 154. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 149, wherein the phospholipid is DOPE.

[0320] Embodiment 155. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 149, wherein the phospholipid is DOPC.

[0321] Embodiment 156. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 155, wherein the sterol is cholesterol, cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2′-hydroxyethyl ether, cholesteryl-4′-hydroxybutyl ether, cholesteryl hemisuccinate, or a mixture of two or more thereof.

[0322] Embodiment 157. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 155, wherein the sterol is cholesterol.

[0323] Embodiment 158. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 157, wherein the polyethylene glycol-lipid conjugate is a polyethylene glycol having a molecular weight from about 1,000 Daltons to about 6,000 Daltons conjugated to a C12–C22 fatty acid lipid.

[0324] Embodiment 159. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 158, wherein the polyethylene glycol has a molecular weight of about 2,000 Daltons.

[0325] Embodiment 160. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 157, wherein the polyethylene glycol-lipid conjugate is DMG-PEG, DPPE-PEG, DPG-PEG, DSG-PEG, DSPE-PEG, or a mixture of two or more thereof; wherein the polyethylene glycol has a molecular weight from about 1,000 Daltons to about 6,000 Daltons.

[0326] Embodiment 161. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 157, wherein the polyethylene glycol-lipid conjugate is DMG-PEG.

[0327] Embodiment 162. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 161, wherein the polyethylene glycol has a molecular weight of about 2,000 Daltons.

[0328] Embodiment 163. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 162, comprising: (i) Lipid 5, Lipid A9, SM-102, ALC-0315, C12- 200, 306-O12B, 4A3-SC8, DODAP, or a combination of two or more thereof; (ii) DPPC, DSPC, DOPE, DOPC, or a combination of two or more thereof; (iii) cholesterol; and (iv) DMG- PEG2000.

[0329] Embodiment 164. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 162, comprising: (i) Lipid 5, Lipid A9, SM-102, ALC-0315, C12- 200, 306-O12B, 4A3-SC8, or DODAP; (ii) DPPC, DSPC, DOPE, or DOPC; (iii) cholesterol; and (iv) DMG-PEG2000.

[0330] Embodiment 165. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising Lipid 5, DPPC, cholesterol, and DMG-PEG2000.

[0331] Embodiment 166. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 165, comprising about 40 mol% to about 60 mol% of Lipid 5, about 5 mol% to about 15 mol% of DPPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000.

[0332] Embodiment 167. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 165, comprising about 50 mol% of Lipid 5, about 10 mol% of DPPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0333] Embodiment 168. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising Lipid A9, DSPC, cholesterol, and DMG-PEG2000.

[0334] Embodiment 169. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 168, comprising about 40 mol% to about 60 mol% of Lipid A9, about 5 mol% to about 15 mol% of DSPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000.

[0335] Embodiment 170. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 168, comprising about 50 mol% of Lipid A9, about 10 mol% of DSPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0336] Embodiment 171. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising SM-102, DSPC, cholesterol, and DMG-PEG2000.

[0337] Embodiment 172. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 171, comprising about 40 mol% to about 60 mol% of SM-102, about 5 mol% to about 15 mol% of DSPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000.

[0338] Embodiment 173. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 171, comprising about 50 mol% of SM-102, about 10 mol% of DSPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0339] Embodiment 174. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising ALC-0315, DSPC, cholesterol, and DMG- PEG2000.

[0340] Embodiment 175. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 174, comprising about 40 mol% to about 60 mol% of ALC-0315, about 5 mol% to about 15 mol% of DSPC, about 36 mol% to about 48 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000.

[0341] Embodiment 176. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 174, comprising about 46.3 mol% of ALC-0315, about 9.4 mol% of DSPC, about 42.7 mol% of cholesterol, and about 1.6 mol% of DMG-PEG2000.

[0342] Embodiment 177. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising C12-200, DOPE, cholesterol, and DMG-PEG2000.

[0343] Embodiment 178. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 177, comprising about 30 mol% to about 40 mol% of C12-200, about 10 mol% to about 20 mol% of DOPE, about 40 mol% to about 50 mol% of cholesterol, and about 1 mol% to about 4 mol% of DMG-PEG2000.

[0344] Embodiment 179. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 177, comprising about 35 mol% of C12-200, about 16 mol% of DOPE, about 46.5 mol% of cholesterol, and about 2.5 mol% of DMG-PEG2000.

[0345] Embodiment 180. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising 306-O12B, DOPC, cholesterol, and DMG- PEG2000.

[0346] Embodiment 181. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 180, comprising about 40 mol% to about 60 mol% of 306-O12B, about 5 mol% to about 15 mol% of DOPC, about 33 mol% to about 44 mol% of cholesterol, and about 0.5 mol% to about 3 mol% of DMG-PEG2000.

[0347] Embodiment 182. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 180, comprising about 50 mol% of 306-O12B, about 10 mol% of DOPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0348] Embodiment 183. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising 4A3-SC8, DOPE, cholesterol, and DMG-PEG2000.

[0349] Embodiment 184. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 183, comprising about 15 mol% to about 30 mol% of 4A3-SC8, about 15 mol% to about 30 mol% of DOPE, about 40 mol% to about 55 mol% of cholesterol, and about 3 mol% to about 7 mol% of DMG-PEG2000.

[0350] Embodiment 185. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 183, comprising about 23.8 mol% of 4A3-SC8, about 23.8 mol% of DOPE, about47.6 mol% of cholesterol, and about 4.8 mol% of DMG-PEG2000.

[0351] Embodiment 186. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising 4A3-SC8, DODAP, DOPE, cholesterol, and DMG- PEG2000.

[0352] Embodiment 187. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 186, comprising about 15 mol% to about 25 mol% of 4A3-SC8, about 15 mol% to about 30 mol% of DODAP, about 15 mol% to about 30 mol% of DOPE, about 30 mol% to about 50 mol% of cholesterol, and about 2 mol% to about 6 mol% of DMG-PEG2000.

[0353] Embodiment 188. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 186, comprising about 19 mol% of 4A3-SC8, about 20 mol% of DODAP, about 19 mol% of DOPE, about 38 mol% of cholesterol, and about 3.8 mol% of DMG-PEG2000.

[0354] Embodiment 189. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 134 to 136, comprising MC3, DSPC, cholesterol, and DMG-PEG2000.

[0355] Embodiment 190. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 189, comprising about 40 mol% to about 60 mol% of MC3, about 5 mol% to about 15 mol% of DSPC, about 33 mol% to about 44 mol% of cholesterol, and about 2 mol% to about 10 mol% of DMG-PEG2000.

[0356] Embodiment 191. The lipid nanoparticle or the plurality of lipid nanoparticles of embodiment 189, comprising about 50 mol% of MC3, about 10 mol% of DSPC, about 38.5 mol% of cholesterol, and about 1.5 mol% of DMG-PEG2000.

[0357] Embodiment 192. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 1 to 191, wherein the lipid nanoparticles have an average size from about 50 nm to about 150 nm.

[0358] Embodiment 193. The lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 1 to 192, wherein the N / P ratio is from about 2:1 to about 8:1.

[0359] Embodiment 194. A pharmaceutical composition comprising the lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 1 to 193 and a pharmaceutically acceptable carrier.

[0360] Embodiment 195. A method of treating or preventing alcohol poisoning in a patient in need thereof, the method comprising administering to the patient an effective amount of the lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 1 to 193 or the pharmaceutical composition of embodiment 194.

[0361] Embodiment 196. A method of treating or preventing a symptom of alcohol poisoning in a patient in need thereof, the method comprising administering to the patient an effectiveamount of the lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 1 to 193 or the pharmaceutical composition of embodiment 194.

[0362] Embodiment 197. The method of embodiment 196, wherein the symptom of alcohol poisoning is mental confusion, hypoglycemia, seizure, slowed breathing, irregular breathing, hypothermia, irregular heartbeat, slow heart rate, loss of consciousness, emesis, heart attack, coma, dehydration, or a combination of two or more thereof.

[0363] Embodiment 198. A method of treating or preventing alcohol flush syndrome in a patient in need thereof, the method comprising administering to the patient an effective amount of the lipid nanoparticle or the plurality of lipid nanoparticles of any one of embodiments 1 to 193 or the pharmaceutical composition of embodiment 194.

[0364] Embodiment 199. The method of any one of embodiments 195 to 198, further comprising administering to the patient an effective amount of oxidized nicotinamide adenine dinucleotide (NAD⁺), a precursor of oxidized nicotinamide adenine dinucleotide (NAD⁺), or a combination thereof.

[0365] Embodiment 200. The method of any one of embodiments 195 to 198, further comprising administering to the patient an effective amount of oxidized nicotinamide adenine dinucleotide (NAD⁺), reduced nicotinamide adenine dinucleotide (NADH), nicotinamide adenine dinucleotide, niacin, nicotinamide, nicotinamide riboside, dihydronicotinamide riboside, nicotinamide mononucleotide, dihydronicotinamide mononucleotide, tryptophan, or a combination of two or more thereof.

[0366] Embodiment 201. The method of any one of embodiments 195 to 198, further comprising administering to the patient an effective amount of niacin, nicotinamide, nicotinamide riboside, nicotinamide mononucleotide, tryptophan, or a combination of two or more thereof.

[0367] Embodiment 202. The method of any one of embodiments 195 to 198, further comprising administering to the patient an effective amount of oxidized nicotinamide adenine dinucleotide (NAD⁺).

[0368] Embodiment 203. The method of any one of embodiments 195-198 and 202, further comprising administering to the patient an effective amount of reduced nicotinamide adenine dinucleotide (NADH).

[0369] Embodiment 204. The method of any one of embodiments 195-198, 202, and 203, further comprising administering to the patient an effective amount of nicotinamide adenine dinucleotide.

[0370] Embodiment 205. The method of any one of embodiments 195-198 and 202-204,further comprising administering to the patient an effective amount of niacin.

[0371] Embodiment 206. The method of any one of embodiments 195-198 and 202-205, further comprising administering to the patient an effective amount of nicotinamide.

[0372] Embodiment 207. The method of any one of embodiments 195-198 and 202-206, further comprising administering to the patient an effective amount of nicotinamide riboside.

[0373] Embodiment 208. The method of any one of embodiments 195-198 and 202-207, further comprising administering to the patient an effective amount of dihydronicotinamide riboside.

[0374] Embodiment 209. The method of any one of embodiments 195-198 and 202-208, further comprising administering to the patient an effective amount of nicotinamide mononucleotide.

[0375] Embodiment 210. The method of any one of embodiments 195-198 and 202-209, further comprising administering to the patient an effective amount of dihydronicotinamide mononucleotide.

[0376] Embodiment 211. The method of any one of embodiments 195-198 and 202-210, further comprising administering to the patient an effective amount of tryptophan.

[0377] Embodiment 212. The method of any one of embodiments 199 to 211, wherein the effective amount produces a plasma concentration from about 0.1 mM to about 20 mM of NAD⁺.

[0378] Embodiment 213. The method of embodiment 212, wherein the effective amount produces a plasma concentration from about 0.1 mM to about 10 mM of NAD⁺.

[0379] Embodiment 214. The method of any one of embodiments 195 to 213, further comprising administering to the patient an effective amount of supplemental magnesium.

[0380] Embodiment 215. A nucleic acid encoding an aldehyde dehydrogenase 2 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:1.

[0381] Embodiment 216. The nucleic acid of embodiment 215 having at least 95% sequence identity to SEQ ID NO:1.

[0382] Embodiment 217. The nucleic acid of embodiment 215 having SEQ ID NO:1.

[0383] Embodiment 218. A nucleic acid encoding an aldehyde dehydrogenase 2 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:2.

[0384] Embodiment 219. The nucleic acid of embodiment 218 having at least 95% sequence identity to SEQ ID NO:2.

[0385] Embodiment 220. The nucleic acid of embodiment 218 having SEQ ID NO:2.

[0386] Embodiment 221. A nucleic acid encoding an aldehyde dehydrogenase 2 enzyme,wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:3.

[0387] Embodiment 222. The nucleic acid of embodiment 221 having at least 95% sequence identity to SEQ ID NO:3.

[0388] Embodiment 223. The nucleic acid of embodiment 221 having SEQ ID NO:3.

[0389] Embodiment 224. A nucleic acid encoding an aldehyde dehydrogenase 2 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:9.

[0390] Embodiment 225. The nucleic acid of embodiment 224 having at least 95% sequence identity to SEQ ID NO:9.

[0391] Embodiment 226. The nucleic acid of embodiment 224 having SEQ ID NO:9.

[0392] Embodiment 227. A nucleic acid encoding an aldehyde dehydrogenase 2 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:10.

[0393] Embodiment 228. The nucleic acid of embodiment 227 having at least 95% sequence identity to SEQ ID NO:10.

[0394] Embodiment 229. The nucleic acid of embodiment 227 having SEQ ID NO:10.

[0395] Embodiment 230. A nucleic acid encoding an aldehyde dehydrogenase 2 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:12.

[0396] Embodiment 231. The nucleic acid of embodiment 230 having at least 95% sequence identity to SEQ ID NO:12.

[0397] Embodiment 232. The nucleic acid of embodiment 230 having SEQ ID NO:12.

[0398] Embodiment 233. A nucleic acid encoding an aldehyde dehydrogenase 2 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:13.

[0399] Embodiment 234. The nucleic acid of embodiment 233 having at least 95% sequence identity to SEQ ID NO:13.

[0400] Embodiment 235. The nucleic acid of embodiment 233 having SEQ ID NO:13.

[0401] Embodiment 236. An aldehyde dehydrogenase 2 enzyme having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:11.

[0402] Embodiment 237. The aldehyde dehydrogenase 2 enzyme of embodiment 236, having an amino acid sequence with at least 95% sequence identity to SEQ ID NO:11.

[0403] Embodiment 238. The aldehyde dehydrogenase 2 enzyme of embodiment 236, comprising SEQ ID NO:11.

[0404] Embodiment 239. An aldehyde dehydrogenase 2 enzyme having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:14.

[0405] Embodiment 240. The aldehyde dehydrogenase 2 enzyme of embodiment 239, having an amino acid sequence with at least 95% sequence identity to SEQ ID NO:14.

[0406] Embodiment 241. The aldehyde dehydrogenase 2 enzyme of embodiment 239, comprising SEQ ID NO:14.

[0407] Embodiment 242. A nucleic acid encoding an alcohol dehydrogenase 1 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:4.

[0408] Embodiment 243. The nucleic acid of embodiment 242 having at least 95% sequence identity to SEQ ID NO:4.

[0409] Embodiment 244. The nucleic acid of embodiment 242 having SEQ ID NO:4.

[0410] Embodiment 245. A nucleic acid encoding an alcohol dehydrogenase 1 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:5.

[0411] Embodiment 246. The nucleic acid of embodiment 245 having at least 95% sequence identity to SEQ ID NO:5.

[0412] Embodiment 247. The nucleic acid of embodiment 245 having SEQ ID NO:5.

[0413] Embodiment 248. A nucleic acid encoding an alcohol dehydrogenase 1 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:6.

[0414] Embodiment 249. The nucleic acid of embodiment 248 having at least 95% sequence identity to SEQ ID NO:6.

[0415] Embodiment 250. The nucleic acid of embodiment 248 having SEQ ID NO:6.

[0416] Embodiment 251. A nucleic acid encoding an alcohol dehydrogenase 1 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:15.

[0417] Embodiment 252. The nucleic acid of embodiment 251 having at least 95% sequence identity to SEQ ID NO:15.

[0418] Embodiment 253. The nucleic acid of embodiment 251 having SEQ ID NO:15.

[0419] Embodiment 254. A nucleic acid encoding an alcohol dehydrogenase 1 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:16.

[0420] Embodiment 255. The nucleic acid of embodiment 254 having at least 95% sequence identity to SEQ ID NO:16.

[0421] Embodiment 256. The nucleic acid of embodiment 254 having SEQ ID NO:16.

[0422] Embodiment 257. A nucleic acid encoding an alcohol dehydrogenase 1 enzyme, wherein the nucleic acid has at least 90% sequence identity to SEQ ID NO:18.

[0423] Embodiment 258. The nucleic acid of embodiment 257 having at least 95% sequence identity to SEQ ID NO:18.

[0424] Embodiment 259. The nucleic acid of embodiment 257 having SEQ ID NO:18.

[0425] Embodiment 260. An alcohol dehydrogenase 1 enzyme having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:17.

[0426] Embodiment 261. The alcohol dehydrogenase 1 enzyme of embodiment 260, having an amino acid sequence with at least 95% sequence identity to SEQ ID NO:17.

[0427] Embodiment 262. The alcohol dehydrogenase 1 enzyme of embodiment 260, comprising SEQ ID NO:17.

[0428] Embodiment 263. An alcohol dehydrogenase 1 enzyme having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:20.

[0429] Embodiment 264. The alcohol dehydrogenase 1 enzyme of embodiment 263, having an amino acid sequence with at least 95% sequence identity to SEQ ID NO:20.

[0430] Embodiment 265. The alcohol dehydrogenase 1 enzyme of embodiment 263, comprising SEQ ID NO:20. EXAMPLES

[0431] The inventors developed an antidote that utilizes in vitro transcribed (IVT) mRNA lipid nanoparticle (LNP) technology to deliver two key alcohol metabolizing enzymes as a treatment for alcohol poisoning. The metabolism of alcohol mainly occurs in the liver by two enzymes, alcohol dehydrogenase (AHD) and aldehyde dehydrogenase (ALDH2). When alcohol (ethanol) is metabolized by these enzymes, ethanol is converted to acetic acid, a relatively non- toxic compound. Patients that have overdosed on alcohol will be treated with IVT mRNA LNP encoding AHD and ALDH2 which would be taken up by hepatocytes in the liver and begin metabolizing alcohol, thus providing a means for rapid clearance of alcohol and an antidote for alcohol poisoning.

[0432] Example 1

[0433] Methods of making lipid nanoparticles (LNPs) are well-known in the art. For example, lipid components are dissolved in ethanol and mixed according to the desired molar ratio (e.g., as described in the Examples herein). The mRNA are dissolved or diluted in acetate or citric buffer at a pH 4-5. NanoAssemblr® Spark™ or Ignite™ systems are used according to manufacturer’s instruction to mix lipid components and mRNA according to the N / P ratio indicated under microfluidic mixing condition to obtain mRNA-LNP. If necessary, dialysis is used to remove ethanol. Encapsulation efficiency is checked using QUANT-IT™ RIBOGREEN® reagent. Particle size is checked using DLS particle sizer.

[0434] Methods for making LNPs are also described, for example, by Cuthbertson et al, “An Introduction to Lipid Nanoparticle Formulation: Basic Concepts & Preparation Procedures,” caymanchem.com / news / intro-to-lipid-nanoparticle-formulation (23 February 2022); and Buschmann et al, “Nanomaterial Delivery Systems for mRNA Vaccines,” Vaccines, 9, 65 (2021).

[0435] Example 2

[0436] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, 19, or 21 will be prepared. The lipid nanoparticles will contain 50 mol% of Lipid 5, 10 mol% of DPPC, 38.5 mol% of cholesterol, and 1.5 mol% of DMG- PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:10 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:16. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:13 and an mRNA having SEQ ID NO:19.

[0437] Example 3

[0438] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 50 mol% of Lipid A9, 10 mol% of DSPC, 38.5 mol% of cholesterol, and 1.5 mol% of DMG- PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:13 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:19. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:10 and an mRNA having SEQ ID NO:16.

[0439] Example 4

[0440] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 50 mol% of SM-102, 10 mol% of DSPC, 38.5 mol% of cholesterol, and 1.5 mol% of DMG- PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:10 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:16. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:13 and an mRNA having SEQ ID NO:19.

[0441] Example 5

[0442] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 46.3 mol% of ALC-0315, 9.4 mol% of DSPC, 42.7 mol% of cholesterol, and 1.6 mol% of DMG- PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:13 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:19. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:10 and an mRNA having SEQ ID NO:16.

[0443] Example 6

[0444] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5,6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 35 mol% of C12-200, 16 mol% of DOPE, 46.5 mol% of cholesterol, and 2.5 mol% of DMG- PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:10 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:16. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:13 and an mRNA having SEQ ID NO:19.

[0445] Example 7

[0446] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 50 mol% of 306-O12B, 10 mol% of DOPC, 38.5 mol% of cholesterol, and 1.5 mol% of DMG- PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:13 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:19. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:10 and an mRNA having SEQ ID NO:16.

[0447] Example 8

[0448] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 23.8 mol% of 4A3-SC8, 23.8 mol% of DOPE, 47.6 mol% of cholesterol, and 4.8 mol% of DMG- PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:10 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:16. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:13 and an mRNA having SEQ ID NO:19.

[0449] Example 9

[0450] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 19 mol% of 4A3-SC8, 20 mol% of DODAP, 19 mol% of DOPE, 38 mol% of cholesterol, and 3.8 mol% of DMG-PEG2000. In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:13 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:19. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:10 and an mRNA having SEQ ID NO:16.

[0451] Example 10

[0452] Lipid nanoparticles containing an mRNA having any one of SEQ ID NO:1, 2, 3, 4, 5, 6, 9, 10, 12, 13, 15, 16, 18, and 19 will be prepared. The lipid nanoparticles will contain 50 mol% of MC3, 10 mol% of DSPC, 38.5 mol% of cholesterol, and 1.5 mol% of DMG-PEG2000.In a first aspect, a first group of lipid nanoparticles will contain an mRNA having SEQ ID NO:10 and a second group of lipid nanoparticles will contain an mRNA having SEQ ID NO:16. In a second aspect, lipid nanoparticles will contain both an mRNA having SEQ ID NO:13 and an mRNA having SEQ ID NO:19.

[0453] Example 11

[0454] The lipid nanoparticles described in Examples 2-10 will be injected into patients that have overdosed on alcohol (e.g., patients having at least one symptom of alcohol overdose and / or a blood alcohol level of 0.30% or more). The treatment will result in a rapid in vivo conversion of alcohol to non-toxic by-products and a successful treatment of alcohol poisoning.

[0455] Nucleic Acids in Examples

[0456] In the Examples herein, the ALDH2*1 (alternatively referred to herein as ALDH2) nucleic acid sequence was SEQ ID NO:2 (wherein the ALDH2*1 nucleic acid sequence including the T7 promoter, poly A tail, and start / stop codons was SEQ ID NO:1) and the ADH1B*2 (alternatively referred to herein as ADH1B) nucleic acid sequence was SEQ ID NO:5 (wherein the ADH1B*2 nucleic acid sequence including the T7 promoter, poly A tail, and start / stop codons was SEQ ID NO:4).

[0457] Example 12

[0458] Preparation of LNP-mRNA. Lipid mixtures (SM102, DSPC, Chol and DMG-PEG2000 at 50:10:38:1.5:0.5 mol ratio) in ethanol was mixed with ALDH2 or ALDH2:ADH at 2:1 ratio in sodium acetate buffer (100 mM, pH 4.0) (N / P ratio at 6) using microfluidic instrument (Precision NanoSystems). Sample was concentrated and buffer exchanged into PBS by Almicon Ultra centrifuge filter right after production. Trapping efficiency was measured by Ribogreen assay. Particle size and zeta potential was measured by DLS instrument. Sample was stored at 4 degree until use. Hassett et al, Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines, Molecular Therapy - Nucleic Acids, 15:1-11 (2019).

[0459] Analytical results of SM102-LNP:mRNA are shown in Table 1 below. mRNA ALDH2 ALDH2:ADH 2:1*

[0460] Entrapment efficiency indicates that mRNA has been trapped inside the lipid nanoparticles to the overall mRNA amount.

[0461] Example 13

[0462] Wt mice C57BL / 6J male mice were treated by retro-orbital injection with: (i) a combination of ALDH2 and ADH (2:1) mRNA-SM102-LNP, (ii) ALDH2 mRNA-SM102-LNP, or (iii) PBS.30 minutes later the mice were administered ethanol at 7g / kg dose by gavage. As shown in FIGS.1A-1B, highly intoxicated mice treated with the combination of ALD / ALDH2 achieved behavioral normalization faster than control. The meaning of the behavior score is shown in Table 2 below.

[0463] Table 2 Behavioral Score for Acute Ethanol Challenge 1 N rm l

[0465] Mice experimental scheme. Control groups (5 mice per group): Injected 100-150ul of PBS or LNP by retro-orbital; Treatment groups (5 mice per group): LNP-ALDH2 mRNA or LNP-ADH1B mRNA diluted in PBS were injected by retro-orbital at 0.25-1 mg / kg dose. After 40 minutes mice received 50% ethanol (diluted in water) at 7g / kg by oral (gavage). Thereafter, mice body temperature and behaviors were observed and scored according to the following system: 1, normal; 2, lethargic (drowsy, doze); 3, mild ataxia (dysfunction of hind limbs); 4, severe ataxia (inability to move); 5, loss of right reflex; 6, dead.24 hours after administration of ethanol mice were euthanized and blood was collected for liver enzyme tests including alanine aminotransferase ALT and aspartate aminotransferase (AST) measurement. Results are shown in FIGS.2A-2C, 3A-3C, 4A-4C, 5A-5C, 6A-6C, 7A-7C, 8A-8B, and 9A-9B.

[0466] Body temperature: Rectal temperature was measured by using Bioseb Thermometer. ALT or AST measurements: measured in serum using the Liquid ALT kit (Pointe Scientific #A7526-01-1953) or Aspartate Aminotransferase colorimetric activity kit (Cayman #701640) following manufacturer’s instructions.

[0467] Materials: Mice: 11-week-old Male C57BL / 6J were purchased from JacksonLaboratory. Ethanol: 200 proof ethanol alcohol from Pharmco AAPER. LNP-ALDH2 mRNA or LNP-ADH1B mRNA made in our laboratory. Preparation of LNP-mRNA. Lipid mixtures (50 mol% of SM-102, 10 mol% of DSPC, 38.5 mol% of cholesterol, and 1.5 mol% of DMG- PEG2000) in ethanol was mixed with ALDH2 or ADH1B at 2:1 ratio in sodium acetate buffer (100 mM, pH 4.0) (N / P ratio at 6) using microfluidic instrument (Precision NanoSystems). Sample was concentrated and buffer exchanged into PBS by Almicon Ultra centrifuge filter right after production. Trapping efficiency was measured by Ribogreen assay. Particle size and zeta potential was measured by DLS instrument. Sample was stored at 4 degree until use.

[0468] Example 14

[0469] ALDH2 Activity Assay. The microplate assay abcam115348 was used to determine mitochondrial aldehyde dehydrogenase activity (ALDH2) in a sample. The enzyme was captured within the cells of the microplate and activity was determined by following the production of NADH in the following ALDH2 catalyzed reaction: acetaldehyde + NAD+= acid + NADH (increased absorbance at 450 nm). The generation of NADH is coupled to the 1:1 reduction of a reporter dye to yield a yellow colored reaction product whose concentration can be monitored by measuring the increase in absorbance at 450 nm (dye molr extinction coefficient is 37000 M-1cm-1). abcam115348 immunocaptures in each well only native ALDH2 from the chosen sample. This removes all other enzymes, including unrelated aldehyde dehydrogenases.

[0470] Samples were prepared as instructed, the protein concentration of extracts was determined, the assay was performed as instructed, and the results were recorded for OD 450 nm for 30-120 minutes. With reference to FIG.10, HeLa cells were grown in 6 well plates and treated with LNP-ALDH2-LNP or LNP-GFP (1 µg mRNA / well) for 20 hours, in which 50 µg protein extract were used per well.

[0471] Lipid mixtures (50 mol% of SM-102, 10 mol% of DSPC, 38.5 mol% of cholesterol, and 1.5 mol% of DMG-PEG2000) in ethanol was mixed with ALDH2 or GFP at 2:1 ratio in sodium acetate buffer (100 mM, pH 4.0) (N / P ratio at 6) using microfluidic instrument (Precision NanoSystems). Sample was concentrated and buffer exchanged into PBS by Almicon Ultra centrifuge filter right after production. Trapping efficiency was measured by Ribogreen assay. Particle size and zeta potential was measured by DLS instrument. Sample was stored at 4 degree until use.

[0472] Example 15

[0473] An ADH activity assay was performed. The alcohol dehydrogenase (ADH) assay kit (MAK498 by Sigma-Aldrich) is based on the reduction of the tetrazolium salt MTT in a NADHcoupled enzymatic reaction to a reduced form of MTT, which exhibits an absorption maximum at 565 nm. The increase in absorbance at 565 nm is directly proportional to the enzyme activity. HeLa cells grown in 6 well plates were transfected with ADH1B*2 or GFP mRNA (1 μgmRNA / well) for 24 h. 50 μg protein extract was used per well. The ADH activity assay wasperformed as instructed, and the OD 565 for 30 minutes was recorded and shown in FIG.11.

[0474] Example 16

[0475] Using AI-assisted tools, the hypothetical rate of increased alcohol metabolism due to the mRNA-LNP described herein (e.g., Examples 12-15) were modeled.

[0476] Assumptions: Delivering codon-optimized mRNA encoding cytosolic ADH1B and mitochondrial ALDH2. The mRNA-LNP is delivered IV (most common for rapid systemic exposure). Liver is targeted, which is responsible for about 90% of ethanol metabolism. Baseline ethanol clearance in an average adult is about 7–10 g / hour, mostly limited by enzyme capacity (zero-order kinetics at high ethanol concentrations).

[0477] Protein Expression Kinetics from mRNA-LNPs. Typical protein expression data from LNP-delivered mRNA in humans (based on vaccine and preclinical therapeutic literature): Peak expression occurs about 6–12 hours post-injection. Protein expression levels can reach micrograms per mL of serum, but tissue-specific delivery (e.g., to hepatocytes) varies. Assuming robust liver targeting, 10x–100x overexpression of a given enzyme would be expected vs baseline for 24–72 hours.

[0478] Enzyme Kinetics. Using a simplified Michaelis-Menten framework for approximation: ADH1B has a VmaxV_{\text{max}}Vmax in liver cytosol that supports about 10 g / hour alcohol metabolism. If mRNA-LNP causes 10x ADH1B expression, the new theoretical VmaxV_{\text{max}}Vmax would be 100 g / hour, assuming NAD⁺ isn’t limiting. This assumes: Sufficient cofactor regeneration (NAD⁺, NADH cycling). ALDH2 isn't the bottleneck. ALDH2 is mitochondrial and handles acetaldehyde: ALDH2*2 mutant carriers (common in people of East Asian descent) have <10% activity, leading to acetaldehyde buildup (flush, toxicity). Restoring ALDH2 activity via mRNA could theoretically normalize or exceed wild-type metabolism, removing this major bottleneck.

[0479] Table 3: Overall Effect on Alcohol Clearance. Given the enzyme overexpressionlevels: Scenario Estimated Clearance RatePractical Estimate 3-5 times faser clearance, possibly up to 10 times depending on dose, delivery and f t li it DH) maybecome limiting unless supported by cofactor regeneration or metabolic pathway flux. Toxicity from excess acetaldehyde could occur if ADH outpaces ALDH2 expression. Immune response to mRNA / LNPs or overexpressed proteins could modulate repeat dosing.

[0481] Summary. Successful expression of functional ADH and ALDH2 in hepatocytes via mRNA-LNP could increase ethanol clearance 3-10x over baseline, thereby reducing intoxication duration from several hours to under one hour, depending on dose. The theoretical results are shown in FIG.12.

[0482] Informal Sequence Listing

[0483] SEQ ID NO:1 = ALDH2*1 –IVT RNA sequence; including T7 promoter, poly A tail, and start / stop codons Start / stop codons are bold and underlined. aggaauaaacuaguauucuucugguccccacagacucagagagaacccgccaccauguugcgcgcugccgcccgcuucgggc cccgccugggccgccgccucuugucagccgccgccacccaggccgugccugcccccaaccagcagcccgaggucuucugcaac cagauuuucauaaacaaugaauggcacgaugccgucagcaggaaaacauuccccaccgucaauccguccacuggagaggucau cugucagguagcugaaggggacaaggaagauguggacaaggcagugaaggccgcccgggccgccuuccagcugggcucaccu uggcgccgcauggacgcaucacacaggggccggcugcugaaccgccuggccgaucugaucgagcgggaccggaccuaccugg cggccuuggagacccuggacaauggcaagcccuaugucaucuccuaccugguggauuuggacaugguccucaaaugucuccg guauuaugccggcugggcugauaaguaccacgggaaaaccauccccauugacggagacuucuucagcuacacacgccaugaa ccugugggggugugcgggcagaucauuccguggaauuucccgcuccugaugcaagcauggaagcugggcccagccuuggca acuggaaacgugguugugaugaagguagcugagcagacaccccucaccgcccucuauguggccaaccugaucaaggaggcug gcuuucccccugguguggucaacauugugccuggauuuggccccacggcuggggccgccauugccucccaugaggaugugg acaaaguggcauucacaggcuccacugagauuggccgcguaauccagguugcugcugggagcagcaaccucaagagagugac cuuggagcugggggggaagagccccaacaucaucaugucagaugccgauauggauugggccguggaacaggcccacuucgcc cuguucuucaaccagggccagugcugcugugccggcucccggaccuucgugcaggaggacaucuaugaugaguuuguggag cggagcguugcccgggccaagucucggguggucgggaaccccuuugauagcaagaccgagcaggggccgcagguggaugaa acucaguuuaagaagauccucggcuacaucaacacggggaagcaagagggggcgaagcugcuguguggugggggcauugcu gcugaccgugguuacuucauccagcccacuguguuuggagaugugcaggauggcaugaccaucgccaaggaggagaucuuc gggccagugaugcagauccugaaguucaagaccauagaggagguuguugggagagccaacaauuccacguacgggcuggccg cagcugucuucacaaaggauuuggacaaggccaauuaccugucccaggcccuccaggcgggcacugugugggucaacugcua ugauguguuuggagcccagucacccuuugguggcuacaagaugucggggaguggccgggaguugggcgaguacgggcugc aggcauacacugaagugaaaacugucacagucaaagugccucagaagaacucauaaugagcuggagccucgguggccuagcuucuugccccuugggccuccccccagccccuccuccccuuccugcacccguacccccguggucuuugaauaaagucugagugg gcggcaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaccagcagcccgaggucuucugcaaccagauuuucauaaacaaugaauggcacgaugccgucagcaggaaaacauuccccacc gucaauccguccacuggagaggucaucugucagguagcugaaggggacaaggaagauguggacaaggcagugaaggccgccc gggccgccuuccagcugggcucaccuuggcgccgcauggacgcaucacacaggggccggcugcugaaccgccuggccgaucu gaucgagcgggaccggaccuaccuggcggccuuggagacccuggacaauggcaagcccuaugucaucuccuaccugguggau uuggacaugguccucaaaugucuccgguauuaugccggcugggcugauaaguaccacgggaaaaccauccccauugacggag acuucuucagcuacacacgccaugaaccugugggggugugcgggcagaucauuccguggaauuucccgcuccugaugcaagc auggaagcugggcccagccuuggcaacuggaaacgugguugugaugaagguagcugagcagacaccccucaccgcccucuau guggccaaccugaucaaggaggcuggcuuucccccugguguggucaacauugugccuggauuuggccccacggcuggggcc gccauugccucccaugaggauguggacaaaguggcauucacaggcuccacugagauuggccgcguaauccagguugcugcug ggagcagcaaccucaagagagugaccuuggagcugggggggaagagccccaacaucaucaugucagaugccgauauggauug ggccguggaacaggcccacuucgcccuguucuucaaccagggccagugcugcugugccggcucccggaccuucgugcaggag gacaucuaugaugaguuuguggagcggagcguugcccgggccaagucucggguggucgggaaccccuuugauagcaagacc gagcaggggccgcagguggaugaaacucaguuuaagaagauccucggcuacaucaacacggggaagcaagagggggcgaagc ugcuguguggugggggcauugcugcugaccgugguuacuucauccagcccacuguguuuggagaugugcaggauggcaug accaucgccaaggaggagaucuucgggccagugaugcagauccugaaguucaagaccauagaggagguuguugggagagcca acaauuccacguacgggcuggccgcagcugucuucacaaaggauuuggacaaggccaauuaccugucccaggcccuccaggc gggcacugugugggucaacugcuaugauguguuuggagcccagucacccuuugguggcuacaagaugucggggaguggccg ggaguugggcgaguacgggcugcaggcauacacugaagugaaaacugucacagucaaagugccucagaagaacuca

[0485] SEQ ID NO:3 = ALDH2*1 IVT vector– complete sequence ctcgagtcgacctaggatcccttctactgggcggttttatggacagcaagcgaaccggaattgccagctggggcgccctctggtaaggttg ggaagccctgcaaagtaaactggatggctttcttgccgccaaggatctgatggcgcaggggatcaagctctgatcaagagacaggatgag gatcgtttcgcatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaaca gacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccggtgccctga atgaactgcaagacgaggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcg ggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccgagaaagtatccatcatggctg atgcaatgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacatcgcatcgagcgagcacgtactcgga tggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgttcgccaggctcaaggcgagc atgcccgacggcgaggatctcgtcgtgacccatggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttctggattcatcga ctgtggccggctgggtgtggcggaccgctatcaggacatagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggctgaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcatcgccttctatcgccttcttgacgagttcttctgaattattaacgcttac aatttcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatcaggtggcacttttcggggaaatgtgcgcggaaccccta tttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatagcacgtgctaaaacttcattttta atttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtaga aaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgc cggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttagg ccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtctt accgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagc gaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatcc ggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgcc acctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctg gccttttgctggccttttgctcacatgttcttgctgcttcgcgatgtacgggccagatatacgcgttgacattgattattgactagttattaatagta atcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacga cccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaa actgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcc cagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaat gggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacggga ctttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctggc taactagagaacccactgcttactggcttatcgaaattaatacgactcactataaggaataaactagtattcttctggtccccacagactcaga gagaacccgccaccatgttgcgcgctgccgcccgcttcgggccccgcctgggccgccgcctcttgtcagccgccgccacccaggccgt gcctgcccccaaccagcagcccgaggtcttctgcaaccagattttcataaacaatgaatggcacgatgccgtcagcaggaaaacattcccc accgtcaatccgtccactggagaggtcatctgtcaggtagctgaaggggacaaggaagatgtggacaaggcagtgaaggccgcccggg ccgccttccagctgggctcaccttggcgccgcatggacgcatcacacaggggccggctgctgaaccgcctggccgatctgatcgagcgg gaccggacctacctggcggccttggagaccctggacaatggcaagccctatgtcatctcctacctggtggatttggacatggtcctcaaatg tctccggtattatgccggctgggctgataagtaccacgggaaaaccatccccattgacggagacttcttcagctacacacgccatgaacctg tgggggtgtgcgggcagatcattccgtggaatttcccgctcctgatgcaagcatggaagctgggcccagccttggcaactggaaacgtgg ttgtgatgaaggtagctgagcagacacccctcaccgccctctatgtggccaacctgatcaaggaggctggctttccccctggtgtggtcaac attgtgcctggatttggccccacggctggggccgccattgcctcccatgaggatgtggacaaagtggcattcacaggctccactgagattg gccgcgtaatccaggttgctgctgggagcagcaacctcaagagagtgaccttggagctgggggggaagagccccaacatcatcatgtca gatgccgatatggattgggccgtggaacaggcccacttcgccctgttcttcaaccagggccagtgctgctgtgccggctcccggaccttcg tgcaggaggacatctatgatgagtttgtggagcggagcgttgcccgggccaagtctcgggtggtcgggaacccctttgatagcaagaccg agcaggggccgcaggtggatgaaactcagtttaagaagatcctcggctacatcaacacggggaagcaagagggggcgaagctgctgtg tggtgggggcattgctgctgaccgtggttacttcatccagcccactgtgtttggagatgtgcaggatggcatgaccatcgccaaggaggag atcttcgggccagtgatgcagatcctgaagttcaagaccatagaggaggttgttgggagagccaacaattccacgtacgggctggccgcagctgtcttcacaaaggatttggacaaggccaattacctgtcccaggccctccaggcgggcactgtgtgggtcaactgctatgatgtgtttgga gcccagtcaccctttggtggctacaagatgtcggggagtggccgggagttgggcgagtacgggctgcaggcatacactgaagtgaaaac tgtcacagtcaaagtgcctcagaagaactcataatgagctggagcctcggtggcctagcttcttgccccttgggcctccccccagcccctcc tccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaagctt

[0486] SEQ ID NO:4 = ADH1B*2 –IVT RNA sequence (including T7 promoter, poly A tail, and start / stop codons). Start / stop codons are bold and underlined. aggaauaaacuaguauucuucugguccccacagacucagagagaacccgccaccaugagcacagcaggaaaaguaaucaaaug caaagcagcugugcuaugggagguaaagaaacccuuuuccauugaggauguggagguugcaccuccuaaggcuuaugaagu ucgcauuaagaugguggcuguaggaaucugucacacagaugaccacgugguuaguggcaaccuggugaccccccuuccugu gauuuuaggccaugaggcagccggcaucguggagaguguuggagaaggggugacuacagucaaaccaggugauaaagucau cccgcucuuuacuccucaguguggaaaaugcagaguuuguaaaaacccggagagcaacuacugcuugaaaaaugaucuaggc aauccucgggggacccugcaggauggcaccaggagguucaccugcagggggaagcccauucaccacuuccuuggcaccagca ccuucucccaguacacggugguggaugagaaugcaguggccaaaauugaugcagccucgccccuggagaaagucugccucau uggcuguggauucucgacugguuaugggucugcaguuaacguugccaaggucaccccaggcucuaccugugcuguguuugg ccugggaggggucggccuaucugcuguuaugggcuguaaagcagcuggagcagccagaaucauugcgguggacaucaacaa ggacaaauuugcaaaggccaaagaguugggugccacugaaugcaucaacccucaagacuacaagaaacccauucaggaagugc uaaaggaaaugacugauggagguguggauuuuucguuugaagucaucggucggcuugacaccaugauggcuucccuguuau guugucaugaggcauguggcacaagcgucaucguagggguaccuccugcuucccagaaccucucaauaaacccuaugcugcu acugacuggacgcaccuggaagggggcuguuuaugguggcuuuaagaguaaagaagguaucccaaaacuuguggcugauuu uauggcuaagaaguuuucacuggaugcguuaauaacccauguuuuaccuuuugaaaaaauaaaugaaggauuugaccugcu ucacucugggaaaaguauccguaccguccugacguuuugaugagcuggagccucgguggccuagcuucuugccccuugggc cuccccccagccccuccuccccuuccugcacccguacccccguggucuuugaauaaagucugagugggcggcaaaaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaagcacagcaggaaaaguaaucaaaugcaaagcagcugugcuaugggagguaaagaaacccuuuuccauugaggauguggagg uugcaccuccuaaggcuuaugaaguucgcauuaagaugguggcuguaggaaucugucacacagaugaccacgugguuagug gcaaccuggugaccccccuuccugugauuuuaggccaugaggcagccggcaucguggagaguguuggagaaggggugacua cagucaaaccaggugauaaagucaucccgcucuuuacuccucaguguggaaaaugcagaguuuguaaaaacccggagagcaa cuacugcuugaaaaaugaucuaggcaauccucgggggacccugcaggauggcaccaggagguucaccugcagggggaagccc auucaccacuuccuuggcaccagcaccuucucccaguacacggugguggaugagaaugcaguggccaaaauugaugcagccu cgccccuggagaaagucugccucauuggcuguggauucucgacugguuaugggucugcaguuaacguugccaaggucaccc caggcucuaccugugcuguguuuggccugggaggggucggccuaucugcuguuaugggcuguaaagcagcuggagcagccagaaucauugcgguggacaucaacaaggacaaauuugcaaaggccaaagaguugggugccacugaaugcaucaacccucaagac uacaagaaacccauucaggaagugcuaaaggaaaugacugauggagguguggauuuuucguuugaagucaucggucggcuu gacaccaugauggcuucccuguuauguugucaugaggcauguggcacaagcgucaucguagggguaccuccugcuucccag aaccucucaauaaacccuaugcugcuacugacuggacgcaccuggaagggggcuguuuaugguggcuuuaagaguaaagaag guaucccaaaacuuguggcugauuuuauggcuaagaaguuuucacuggaugcguuaauaacccauguuuuaccuuuugaaa aaauaaaugaaggauuugaccugcuucacucugggaaaaguauccguaccguccugacguuuuga

[0488] SEQ ID NO:6 = ADH1B*2 IVT vector - complete sequence ctcgagtcgacctaggatcccttctactgggcggttttatggacagcaagcgaaccggaattgccagctggggcgccctctggtaaggttg ggaagccctgcaaagtaaactggatggctttcttgccgccaaggatctgatggcgcaggggatcaagctctgatcaagagacaggatgag gatcgtttcgcatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaaca gacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccggtgccctga atgaactgcaagacgaggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcg ggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccgagaaagtatccatcatggctg atgcaatgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacatcgcatcgagcgagcacgtactcgga tggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgttcgccaggctcaaggcgagc atgcccgacggcgaggatctcgtcgtgacccatggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttctggattcatcga ctgtggccggctgggtgtggcggaccgctatcaggacatagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggctga ccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcatcgccttctatcgccttcttgacgagttcttctgaattattaacgcttac aatttcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatcaggtggcacttttcggggaaatgtgcgcggaaccccta tttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatagcacgtgctaaaacttcattttta atttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtaga aaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgc cggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttagg ccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtctt accgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagc gaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatcc ggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgcc acctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctg gccttttgctggccttttgctcacatgttcttgctgcttcgcgatgtacgggccagatatacgcgttgacattgattattgactagttattaatagta atcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacga cccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaa actgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcc cagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaat gggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctggc taactagagaacccactgcttactggcttatcgaaattaatacgactcactataaggaataaactagtattcttctggtccccacagactcaga gagaacccgccaccatgagcacagcaggaaaagtaatcaaatgcaaagcagctgtgctatgggaggtaaagaaacccttttccattgagg atgtggaggttgcacctcctaaggcttatgaagttcgcattaagatggtggctgtaggaatctgtcacacagatgaccacgtggttagtggca acctggtgaccccccttcctgtgattttaggccatgaggcagccggcatcgtggagagtgttggagaaggggtgactacagtcaaaccag gtgataaagtcatcccgctctttactcctcagtgtggaaaatgcagagtttgtaaaaacccggagagcaactactgcttgaaaaatgatctag gcaatcctcgggggaccctgcaggatggcaccaggaggttcacctgcagggggaagcccattcaccacttccttggcaccagcaccttct cccagtacacggtggtggatgagaatgcagtggccaaaattgatgcagcctcgcccctggagaaagtctgcctcattggctgtggattctc gactggttatgggtctgcagttaacgttgccaaggtcaccccaggctctacctgtgctgtgtttggcctgggaggggtcggcctatctgctgtt atgggctgtaaagcagctggagcagccagaatcattgcggtggacatcaacaaggacaaatttgcaaaggccaaagagttgggtgccact gaatgcatcaaccctcaagactacaagaaacccattcaggaagtgctaaaggaaatgactgatggaggtgtggatttttcgtttgaagtcatc ggtcggcttgacaccatgatggcttccctgttatgttgtcatgaggcatgtggcacaagcgtcatcgtaggggtacctcctgcttcccagaac ctctcaataaaccctatgctgctactgactggacgcacctggaagggggctgtttatggtggctttaagagtaaagaaggtatcccaaaactt gtggctgattttatggctaagaagttttcactggatgcgttaataacccatgttttaccttttgaaaaaataaatgaaggatttgacctgcttcactc tgggaaaagtatccgtaccgtcctgacgttttgatgagctggagcctcggtggcctagcttcttgccccttgggcctccccccagcccctcct ccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaagctt

[0489] SEQ ID NO: 7 = aldehyde dehydrogenase 2 enzyme MLRAAARFGPRLGRRLLSAAATQAVPAPNQQPEVFCNQIFINNEWHDAVSRKTFPTVNPS TGEVICQVAEGDKEDVDKAVKAARAAFQLGSPWRRMDASHRGRLLNRLADLIERDRTYLA ALETLDNGKPYVISYLVDLDMVLKCLRYYAGWADKYHGKTIPIDGDFFSYTRHEPVGVCG QIIPWNFPLLMQAWKLGPALATGNVVVMKVAEQTPLTALYVANLIKEAGFPPGVVNIVPG FGPTAGAAIASHEDVDKVAFTGSTEIGRVIQVAAGSSNLKRVTLELGGKSPNIIMSDADM DWAVEQAHFALFFNQGQCCCAGSRTFVQEDIYDEFVERSVARAKSRVVGNPFDSKTEQGP QVDETQFKKILGYINTGKQEGAKLLCGGGIAADRGYFIQPTVFGDVQDGMTIAKEEIFGP VMQILKFKTIEEVVGRANNSTYGLAAAVFTKDLDKANYLSQALQAGTVWVNCYDVFGAQS PFGGYKMSGSGRELGEYGLQAYTEVKTVTVKVPQKNS

[0490] SEQ ID NO:8 = alcohol dehydrogenase 1 enzyme MSTAGKVIKCKAAVLWELKKPFSIEEVEVAPPKAHEVRIKMVAVGICGTDDHVVSGTMVT PLPVILGHEAAGIVESVGEGVTTVKPGDKVIPLAIPQCGKCRICKNPESNYCLKNDVSNP QGTLQDGTSRFTCRRKPIHHFLGISTFSQYTVVDENAVAKIDAASPLEKVCLIGCGFSTG YGSAVNVAKVTPGSTCAVFGLGGVGLSAIMGCKAAGAARIIAVDINKDKFAKAKELGATE CINPQDYKKPIQEVLKEMTDGGVDFSFEVIGRLDTMMASLLCCHEACGTSVIVGVPPDSQ NLSMNPMLLLTGRTWKGAILGGFKSKECVPKLVADFMAKKFSLDALITHVLPFEKINEGFDLLHSGKSIRTILMF

[0491] SEQ ID NO:9 = ALDH2 – (bold underlined lower case is a start / stop codon). aggaauaaacuaguauucuucugguccccacagacucagagagaacccgccaccauguugcgcgcugccgcccgcuucgggc cccgccugggccgccgccucuugucagccgccgccacccaggccgugccugcccccaaccagcagcccgaggucuucugcaac cagauuuucauaaacaaugaauggcacgaugccgucagcaggaaaacauuccccaccgucaauccguccacuggagaggucau cugucagguagcugaaggggacaaggaagauguggacaaggcagugaaggccgcccgggccgccuuccagcugggcucaccu uggcgccgcauggacgcaucacacaggggccggcugcugaaccgccuggccgaucugaucgagcgggaccggaccuaccugg cggccuuggagacccuggacaauggcaagcccuaugucaucuccuaccugguggauuuggacaugguccucaaaugucuccg guauuaugccggcugggcugauaaguaccacgggaaaaccauccccauugacggagacuucuucagcuacacacgccaugaa ccugugggggugugcgggcagaucauuccguggaauuucccgcuccugaugcaagcauggaagcugggcccagccuuggca acuggaaacgugguugugaugaagguagcugagcagacaccccucaccgcccucuauguggccaaccugaucaaggaggcug gcuuucccccugguguggucaacauugugccuggauuuggccccacggcuggggccgccauugccucccaugaggaugugg acaaaguggcauucacaggcuccacugagauuggccgcguaauccagguugcugcugggagcagcaaccucaagagagugac cuuggagcugggggggaagagccccaacaucaucaugucagaugccgauauggauugggccguggaacaggcccacuucgcc cuguucuucaaccagggccagugcugcugugccggcucccggaccuucgugcaggaggacaucuaugaugaguuuguggag cggagcguugcccgggccaagucucggguggucgggaaccccuuugauagcaagaccgagcaggggccgcagguggaugaa acucaguuuaagaagauccucggcuacaucaacacggggaagcaagagggggcgaagcugcuguguggugggggcauugcu gcugaccgugguuacuucauccagcccacuguguuuggagaugugcaggauggcaugaccaucgccaaggaggagaucuuc gggccagugaugcagauccugaaguucaagaccauagaggagguuguugggagagccaacaauuccacguacgggcuggccg cagcugucuucacaaaggauuuggacaaggccaauuaccugucccaggcccuccaggcgggcacugugugggucaacugcua ugauguguuuggagcccagucacccuuugguggcuacaagaugucggggaguggccgggaguugggcgaguacgggcugc aggcauacacuX1X2X3gugaaaacugucacagucaaagugccucagaagaacucauaaugagcuggagccucgguggccu agcuucuugccccuugggccuccccccagccccuccuccccuuccugcacccguacccccguggucuuugaauaaagucuga gugggcggcaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaaaaa; wherein X1, X2, and X3are each independently a, u, g, or c. In embodiments, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon thatencodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V.

[0492] SEQ ID NO:10 = ALDH2 – IVT RNA sequence uugcgcgcugccgcccgcuucgggccccgccugggccgccgccucuugucagccgccgccacccaggccgugccugccccca accagcagcccgaggucuucugcaaccagauuuucauaaacaaugaauggcacgaugccgucagcaggaaaacauuccccacc gucaauccguccacuggagaggucaucugucagguagcugaaggggacaaggaagauguggacaaggcagugaaggccgccc gggccgccuuccagcugggcucaccuuggcgccgcauggacgcaucacacaggggccggcugcugaaccgccuggccgaucu gaucgagcgggaccggaccuaccuggcggccuuggagacccuggacaauggcaagcccuaugucaucuccuaccugguggau uuggacaugguccucaaaugucuccgguauuaugccggcugggcugauaaguaccacgggaaaaccauccccauugacggag acuucuucagcuacacacgccaugaaccugugggggugugcgggcagaucauuccguggaauuucccgcuccugaugcaagc auggaagcugggcccagccuuggcaacuggaaacgugguugugaugaagguagcugagcagacaccccucaccgcccucuau guggccaaccugaucaaggaggcuggcuuucccccugguguggucaacauugugccuggauuuggccccacggcuggggcc gccauugccucccaugaggauguggacaaaguggcauucacaggcuccacugagauuggccgcguaauccagguugcugcug ggagcagcaaccucaagagagugaccuuggagcugggggggaagagccccaacaucaucaugucagaugccgauauggauug ggccguggaacaggcccacuucgcccuguucuucaaccagggccagugcugcugugccggcucccggaccuucgugcaggag gacaucuaugaugaguuuguggagcggagcguugcccgggccaagucucggguggucgggaaccccuuugauagcaagacc gagcaggggccgcagguggaugaaacucaguuuaagaagauccucggcuacaucaacacggggaagcaagagggggcgaagc ugcuguguggugggggcauugcugcugaccgugguuacuucauccagcccacuguguuuggagaugugcaggauggcaug accaucgccaaggaggagaucuucgggccagugaugcagauccugaaguucaagaccauagaggagguuguugggagagcca acaauuccacguacgggcuggccgcagcugucuucacaaaggauuuggacaaggccaauuaccugucccaggcccuccaggc gggcacugugugggucaacugcuaugauguguuuggagcccagucacccuuugguggcuacaagaugucggggaguggccg ggaguugggcgaguacgggcugcaggcauacacuX1X2X3gugaaaacugucacagucaaagugccucagaagaacuca; wherein X1, X2, and X3are each independently a, u, g, or c. In embodiments, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments,X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V.

[0493] SEQ ID NO:11 – ALDH2 amino acid sequence MLRAAARFGPRLGRRLLSAAATQAVPAPNQQPEVFCNQIFINNEWHDAVSRKTFPTVN PSTGEVICQVAEGDKEDVDKAVKAARAAFQLGSPWRRMDASHRGRLLNRLADLIERD RTYLAALETLDNGKPYVISYLVDLDMVLKCLRYYAGWADKYHGKTIPIDGDFFSYTRH EPVGVCGQIIPWNFPLLMQAWKLGPALATGNVVVMKVAEQTPLTALYVANLIKEAGFP PGVVNIVPGFGPTAGAAIASHEDVDKVAFTGSTEIGRVIQVAAGSSNLKRVTLELGGKSP NIIMSDADMDWAVEQAHFALFFNQGQCCCAGSRTFVQEDIYDEFVERSVARAKSRVVG NPFDSKTEQGPQVDETQFKKILGYINTGKQEGAKLLCGGGIAADRGYFIQPTVFGDVQD GMTIAKEEIFGPVMQILKFKTIEEVVGRANNSTYGLAAAVFTKDLDKANYLSQALQAGT VWVNCYDVFGAQSPFGGYKMSGSGRELGEYGLQAYTX4VKTVTVKVPQKNS; wherein X4is E, S, N, D, A, R, C, Q, G, P, Y, H, I, L, K, M, F, T, W, or V. In embodiments, X4is E. In embodiments, X4is S. In embodiments, X4is N. In embodiments, X4is D. In embodiments, X4is A. In embodiments, X4is R. In embodiments, X4is C. In embodiments, X4is Q. In embodiments, X4is G. In embodiments, X4is P. In embodiments, X4is Y. In embodiments, X4is H. In embodiments, X4is I. In embodiments, X4is L. In embodiments, X4is K. In embodiments, X4is M. In embodiments, X4is F. In embodiments, X4is T. In embodiments, X4is W. In embodiments, X4is V.

[0494] SEQ ID NO:12 = ALDH2 (bold underlined lower case is a start / stop codon). aggaauaaacuaguauucuucugguccccacagacucagagagaacccgccaccaugUUGCGCGCUGCCGCCCG CUUCGGGCCCCGCCUGGGCCGCCGCCUCUUGUCAGCCGCCGCCACCCAGGCCGUG CCUGCCCCCAACCAGCAGCCCGAGGUCUUCUGCAACCAGAUUUUCAUAAACAAUG AAUGGCACGAUGCCGUCAGCAGGAAAACAUUCCCCACCGUCAAUCCGUCCACUGG AGAGGUCAUCUGUCAGGUAGCUGAAGGGGACAAGGCCUUGGAGACCCUGGACAA UGGCAAGCCCUAUGUCAUCUCCUACCUGGUGGAUUUGGACAUGGUCCUCAAAUG UCUCCGGUAUUAUGCCGGCUGGGCUGAUAAGUACCACGGGAAAACCAUCCCCAU UGACGGAGACUUCUUCAGCUACACACGCCAUGAACCUGUGGGGGUGUGCGGGCA GAUCAUUCCGUGGAAUUUCCCGCUCCUGAUGCAAGCAUGGAAGCUGGGCCCAGCC UUGGCAACUGGAAACGUGGUUGUGAUGAAGGUAGCUGAGCAGACACCCCUCACC GCCCUCUAUGUGGCCAACCUGAUCAAGGAGGCUGGCUUUCCCCCUGGUGUGGUCA ACAUUGUGCCUGGAUUUGGCCCCACGGCUGGGGCCGCCAUUGCCUCCCAUGAGGA UGUGGACAAAGUGGCAUUCACAGGCUCCACUGAGAUUGGCCGCGUAAUCCAGGU UGCUGCUGGGAGCAGCAACCUCAAGAGAGUGACCUUGGAGCUGGGGGGGAAGAG CCCCAACAUCAUCAUGUCAGAUGCCGAUAUGGAUUGGGCCGUGGAACAGGCCCACUUCGCCCUGUUCUUCAACCAGGGCCAGUGCUGCUGUGCCGGCUCCCGGACCUUCG UGCAGGAGGACAUCUAUGAUGAGUUUGUGGAGCGGAGCGUUGCCCGGGCCAAGU CUCGGGUGGUCGGGAACCCCUUUGAUAGCAAGACCGAGCAGGGGCCGCAGGUGG AUGAAACUCAGUUUAAGAAGAUCCUCGGCUACAUCAACACGGGGAAGCAAGAGG GGGCGAAGCUGCUGUGUGGUGGGGGCAUUGCUGCUGACCGUGGUUACUUCAUCC AGCCCACUGUGUUUGGAGAUGUGCAGGAUGGCAUGACCAUCGCCAAGGAGGAGA UCUUCGGGCCAGUGAUGCAGAUCCUGAAGUUCAAGACCAUAGAGGAGGUUGUUG GGAGAGCCAACAAUUCCACGUACGGGCUGGCCGCAGCUGUCUUCACAAAGGAUU UGGACAAGGCCAAUUACCUGUCCCAGGCCCUCCAGGCGGGCACUGUGUGGGUCAA CUGCUAUGAUGUGUUUGGAGCCCAGUCACCCUUUGGUGGCUACAAGAUGUCGGG GAGUGGCCGGGAGUUGGGCGAGUACGGGCUGCAGGCAUACACU X1X2X3GUGAAAACUGUCACAGUCAAAGUGCCUCAGAAGAACUCAuaaugagcuggagccu cgguggccuagcuucuugccccuugggccuccccccagccccuccuccccuuccugcacccguacccccguggucuuugaau aaagucugagugggcggcaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa; wherein X1, X2, and X3are each independently a, u, g, or c. In embodiments, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V.

[0495] SEQ ID NO:13 = ALDH2 – nucleotide sequence UUGCGCGCUGCCGCCCGCUUCGGGCCCCGCCUGGGCCGCCGCCUCUUGUCAGCCG CCGCCACCCAGGCCGUGCCUGCCCCCAACCAGCAGCCCGAGGUCUUCUGCAACCA GAUUUUCAUAAACAAUGAAUGGCACGAUGCCGUCAGCAGGAAAACAUUCCCCAC CGUCAAUCCGUCCACUGGAGAGGUCAUCUGUCAGGUAGCUGAAGGGGACAAGGC CUUGGAGACCCUGGACAAUGGCAAGCCCUAUGUCAUCUCCUACCUGGUGGAUUU GGACAUGGUCCUCAAAUGUCUCCGGUAUUAUGCCGGCUGGGCUGAUAAGUACCACGGGAAAACCAUCCCCAUUGACGGAGACUUCUUCAGCUACACACGCCAUGAACCU GUGGGGGUGUGCGGGCAGAUCAUUCCGUGGAAUUUCCCGCUCCUGAUGCAAGCA UGGAAGCUGGGCCCAGCCUUGGCAACUGGAAACGUGGUUGUGAUGAAGGUAGCU GAGCAGACACCCCUCACCGCCCUCUAUGUGGCCAACCUGAUCAAGGAGGCUGGCU UUCCCCCUGGUGUGGUCAACAUUGUGCCUGGAUUUGGCCCCACGGCUGGGGCCGC CAUUGCCUCCCAUGAGGAUGUGGACAAAGUGGCAUUCACAGGCUCCACUGAGAU UGGCCGCGUAAUCCAGGUUGCUGCUGGGAGCAGCAACCUCAAGAGAGUGACCUU GGAGCUGGGGGGGAAGAGCCCCAACAUCAUCAUGUCAGAUGCCGAUAUGGAUUG GGCCGUGGAACAGGCCCACUUCGCCCUGUUCUUCAACCAGGGCCAGUGCUGCUGU GCCGGCUCCCGGACCUUCGUGCAGGAGGACAUCUAUGAUGAGUUUGUGGAGCGG AGCGUUGCCCGGGCCAAGUCUCGGGUGGUCGGGAACCCCUUUGAUAGCAAGACC GAGCAGGGGCCGCAGGUGGAUGAAACUCAGUUUAAGAAGAUCCUCGGCUACAUC AACACGGGGAAGCAAGAGGGGGCGAAGCUGCUGUGUGGUGGGGGCAUUGCUGCU GACCGUGGUUACUUCAUCCAGCCCACUGUGUUUGGAGAUGUGCAGGAUGGCAUG ACCAUCGCCAAGGAGGAGAUCUUCGGGCCAGUGAUGCAGAUCCUGAAGUUCAAG ACCAUAGAGGAGGUUGUUGGGAGAGCCAACAAUUCCACGUACGGGCUGGCCGCA GCUGUCUUCACAAAGGAUUUGGACAAGGCCAAUUACCUGUCCCAGGCCCUCCAGG CGGGCACUGUGUGGGUCAACUGCUAUGAUGUGUUUGGAGCCCAGUCACCCUUUG GUGGCUACAAGAUGUCGGGGAGUGGCCGGGAGUUGGGCGAGUACGGGCUGCAGG CAUACACUX1X2X3GUGAAAACUGUCACAGUCAAAGUGCCUCAGAAGAACUCA; wherein X1, X2, and X3are each independently a, u, g, or c. In embodiments, X1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. In embodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V.

[0496] SEQ ID NO:14 = ALDH2 – amino acid sequence MLRAAARFGPRLGRRLLSAAATQAVPAPNQQPEVFCNQIFINNEWHDAVSRKTFPTVNPSTGEVICQVAEGDKALETLDNGKPYVISYLVDLDMVLKCLRYYAGWADKYHGKTIPI DGDFFSYTRHEPVGVCGQIIPWNFPLLMQAWKLGPALATGNVVVMKVAEQTPLTALY VANLIKEAGFPPGVVNIVPGFGPTAGAAIASHEDVDKVAFTGSTEIGRVIQVAAGSSNLK RVTLELGGKSPNIIMSDADMDWAVEQAHFALFFNQGQCCCAGSRTFVQEDIYDEFVER SVARAKSRVVGNPFDSKTEQGPQVDETQFKKILGYINTGKQEGAKLLCGGGIAADRGY FIQPTVFGDVQDGMTIAKEEIFGPVMQILKFKTIEEVVGRANNSTYGLAAAVFTKDLDK ANYLSQALQAGTVWVNCYDVFGAQSPFGGYKMSGSGRELGEYGLQAYTX5VKTVTVK VPQKNS; wherein X5is E, S, N, D, A, R, C, Q, G, P, Y, H, I, L, K, M, F, T, W, or V. In embodiments, X5is E. In embodiments, X5is S. In embodiments, X5is N. In embodiments, X5is D. In embodiments, X5is A. In embodiments, X5is R. In embodiments, X5is C. In embodiments, X5is Q. In embodiments, X5is G. In embodiments, X5is P. In embodiments, X5is Y. In embodiments, X5is H. In embodiments, X5is I. In embodiments, X5is L. In embodiments, X5is K. In embodiments, X5is M. In embodiments, X5is F. In embodiments, X5is T. In embodiments, X5is W. In embodiments, X5is V.

[0497] SEQ ID NO:15 = ADH1B (bold underlined lower case is a start / stop codon). aggaauaaacuaguauucuucugguccccacagacucagagagaacccgccaccaugagcacagcaggaaaaguaaucaaaug caaagcagcugugcuaugggagguaaagaaacccuuuuccauugaggauguggagguugcaccuccuaaggcuuaugaagu ucgcauuaagaugguggcuguaggaaucuguX1X2X3acagaugaccacgugguuaguggcaaccuggugaccccccuucc ugugauuuuaggccaugaggcagccggcaucguggagaguguuggagaaggggugacuacagucaaaccaggugauaaagu caucccgcucuuuacuccucaguguggaaaaugcagaguuuguaaaaacccggagagcaacuacugcuugaaaaaugaucua ggcaauccucgggggacccugcaggauggcaccaggagguucaccugcagggggaagcccauucaccacuuccuuggcacca gcaccuucucccaguacacggugguggaugagaaugcaguggccaaaauugaugcagccucgccccuggagaaagucugccu cauuggcuguggauucucgacugguuaugggucugcaguuaacguugccaaggucaccccaggcucuaccugugcuguguu uggccugggaggggucggccuaucugcuguuaugggcuguaaagcagcuggagcagccagaaucauugcgguggacaucaa caaggacaaauuugcaaaggccaaagaguugggugccacugaaugcaucaacccucaagacuacaagaaacccauucaggaag ugcuaaaggaaaugacugauggagguguggauuuuucguuugaagucaucggucggcuugacaccaugauggcuucccugu uauguugucaugaggcauguggcacaagcgucaucguagggguaccuccugcuucccagaaccucucaauaaacccuaugcu gcuacugacuggacgcaccuggaagggggcuguuuaugguggcuuuaagaguaaagaagguaucccaaaacuuguggcuga uuuuauggcuaagaaguuuucacuggaugcguuaauaacccauguuuuaccuuuugaaaaaauaaaugaaggauuugaccu gcuucacucugggaaaaguauccguaccguccugacguuuugaugagcuggagccucgguggccuagcuucuugccccuug ggccuccccccagccccuccuccccuuccugcacccguacccccguggucuuugaauaaagucugagugggcggcaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaX1X2X3is a codon that encodes E. In embodiments, X1X2X3is a codon that encodes S. Inembodiments, X1X2X3is a codon that encodes N. In embodiments, X1X2X3is a codon that encodes D. In embodiments, X1X2X3is a codon that encodes A. In embodiments, X1X2X3is a codon that encodes R. In embodiments, X1X2X3is a codon that encodes C. In embodiments, X1X2X3is a codon that encodes Q. In embodiments, X1X2X3is a codon that encodes G. In embodiments, X1X2X3is a codon that encodes P. In embodiments, X1X2X3is a codon that encodes Y. In embodiments, X1X2X3is a codon that encodes H. In embodiments, X1X2X3is a codon that encodes I. In embodiments, X1X2X3is a codon that encodes L. In embodiments, X1X2X3is a codon that encodes K. In embodiments, X1X2X3is a codon that encodes M. In embodiments, X1X2X3is a codon that encodes F. In embodiments, X1X2X3is a codon that encodes T. In embodiments, X1X2X3is a codon that encodes W. In embodiments, X1X2X3is a codon that encodes V.

[0498] SEQ ID NO:16 = ADH1B – nucleotide sequence agcacagcaggaaaaguaaucaaaugcaaagcagcugugcuaugggagguaaagaaacccuuuuccauugaggauguggagg uugcaccuccuaaggcuuaugaaguucgcauuaagaugguggcuguaggaaucuguX1X2X3acagaugaccacgugguua guggcaaccuggugaccccccuuccugugauuuuaggccaugaggcagccggcaucguggagaguguuggagaagggguga cuacagucaaaccaggugauaaagucaucccgcucuuuacuccucaguguggaaaaugcagaguuuguaaaaacccggagag caacuacugcuugaaaaaugaucuaggcaauccucggggga...

Claims

CLAIMS What is claimed is:

1. A lipid nanoparticle comprising: (i) a nucleic acid encoding an aldehyde dehydrogenase 2 enzyme, (ii) a nucleic acid encoding an alcohol dehydrogenase 1 enzyme, (iii) a nucleic acid encoding an aldehyde dehydrogenase 2 enzyme and an alcohol dehydrogenase 1 enzyme, or (iv) a first nucleic acid encoding an aldehyde dehydrogenase 2 enzyme and a second nucleic acid encoding a alcohol dehydrogenase 1 enzyme.

2. The lipid nanoparticle of claim 1, wherein: (a) the nucleic acid encoding the aldehyde dehydrogenase 2 enzyme has at least 90% sequence identity to SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:21, SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO:12, and (b) the nucleic acid encoding the alcohol dehydrogenase 1 enzyme has at least 90% sequence identity to SEQ ID NO:5, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:15, or SEQ ID NO:

18.

3. The lipid nanoparticle of claim 1, wherein the nucleic acid is RNA.

4. A lipid nanoparticle comprising: (i) an aldehyde dehydrogenase 2 enzyme, (ii) an alcohol dehydrogenase 1 enzyme, (iii) an aldehyde dehydrogenase 2 enzyme and an alcohol dehydrogenase 1 enzyme.

5. The lipid nanoparticle of claim 1, wherein: (a) the aldehyde dehydrogenase 2 enzyme has at least 90% sequence identity to SEQ ID NO:11, or SEQ ID NO:14, and (b) the alcohol dehydrogenase 1 enzyme has at least 90% sequence identity to SEQ ID NO:17 or SEQ ID NO:

20.

6. A plurality of lipid nanoparticles comprising the lipid nanoparticle of claim 1.

7. A plurality of lipid nanoparticles comprising: (i) a first lipid nanoparticle comprising a first nucleic acid encoding an aldehyde dehydrogenase 2 enzyme; and (ii) a second lipid nanoparticle comprising a second nucleic acid encoding an alcohol dehydrogenase 1 enzyme.

8. The lipid nanoparticles of claim 7, wherein the ratio of the first lipid nanoparticles to the second lipid nanoparticles is from 95:5 to 5:

95.

9. A plurality of lipid nanoparticles comprising: (i) a first lipid nanoparticle comprising an aldehyde dehydrogenase 2 enzyme; and (ii) a second lipid nanoparticle comprising an alcohol dehydrogenase 1 enzyme.

10. The lipid nanoparticle of claim 1 comprising: (i) a cationic lipid: (ii) a phospholipid; (iii) a sterol; and (iv) a polyethylene glycol-lipid conjugate.

11. The lipid nanoparticle of claim 10, comprising: (i) about 30 mole% to about 70 mole% of the cationic lipid: (ii) about 5 mole% to about 20 mole% of the phospholipid; (iii) about 20 mole% to 50 mole% of the sterol; and (iv) about 0.1 mole% to about 10 mole% of the polyethylene glycol-lipid conjugate.

12. The lipid nanoparticle of claim 10, comprising: (i) about 10 mole% to about 40 mole% of the cationic lipid: (ii) about 15 mole% to about 50 mole% of the phospholipid; (iii) about 30 mole% to 60 mole% of the sterol; and (iv) about 0.1 mole% to about 10 mole% of the polyethylene glycol-lipid conjugate.

13. The lipid nanoparticle of claim 10, wherein:(i) the cationic lipid is DOTAP, DODAC, DODMA, DSDMA, DOTMA, DDAB, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DOAP, C12-200, 98N12-5, Lipid 5, Lipid A9, 4A3-SC8, 306-012B, DODAP, SM- 102, ALC-0315, L-319, 5A2-SC8, 306Oi10, Lipid CL1, ATX-0114, MC3, an MC3 derivative, DLinDMA, DLin-K-C2-DMA, DLin-K-C3-DMA, DLin-K-C4-DMA, DLin-K6-DMA, DLin-K-MPZ, DLin-K-DMA, DLin-C-DAP, DLin-DAC, DLin- MA, DLinDAP, DLin-S-DMA, DLin-2-DMAP, DLin-TMA, DLin-TAP, DLin- MPZ, DLinAP, DLin-EG-DMA, DLincarbDAP, or a mixture of two or more thereof;(ii) the phospholipid is DSPC, DPPC, DOPE, POPC, POPE, POPG, DPPE, DMPE,DSPE, MMPE, DMPE, DEPE, SOPE, EPC, HSPC, DPPG, or a mixture of two or more thereof;(iii) the sterol is cholesterol, cholestanol, cholestanone, cholestenone, coprostanol,cholesteryl-2′-hydroxyethyl ether, cholesteryl-4′-hydroxybutyl ether, cholesteryl hemisuccinate, or a mixture of two or more thereof; and(iv) the polyethylene glycol-lipid conjugate is a polyethylene glycol having a molecularweight from about 1,000 Daltons to about 6,000 Daltons conjugated to a C12-C22fatty acid lipid.

14. The lipid nanoparticle of claim 10, comprising: (i) Lipid 5, Lipid A9, SM-102, ALC-0315, C12-200, 306-O12B, 4A3-SC8, DODAP, or a combination of two or more thereof; (ii) DPPC, DSPC, DOPE, DOPC, or a combination of two or more thereof; (iii) cholesterol; and (iv) DMG-PEG2000.

15. The lipid nanoparticle of claim 10, comprising: (a) SM-102, DSPC, cholesterol, and DMG-PEG2000,(b) Lipid 5, DPPC, cholesterol, and DMG-PEG2000,(c) Lipid A9, DSPC, cholesterol, and DMG-PEG2000,(d) ALC-0315, DSPC, cholesterol, and DMG-PEG2000, (e) C12-200, DOPE, cholesterol, and DMG-PEG2000, (f) 306-O12B, DOPC, cholesterol, and DMG-PEG2000, (g) 4A3-SC8, DOPE, cholesterol, and DMG-PEG2000,(h) 4A3-SC8, DODAP, DOPE, cholesterol, and DMG-PEG2000, or (i) MC3, DSPC, cholesterol, and DMG-PEG2000.

16. The lipid nanoparticle of claim 1, wherein the lipid nanoparticles have an average size from about 50 nm to about 150 nm.

17. The lipid nanoparticle of claim 1, wherein the N / P ratio is from about 2:1 to about 8:

1.

18. A pharmaceutical composition comprising the lipid nanoparticle of claim 1 and a pharmaceutically acceptable carrier.

19. A method of treating or preventing alcohol poisoning, treating or preventing a symptom of alcohol poisoning, or treating or preventing alcohol flush syndrome in a patient in need thereof, the method comprising administering to the patient an effective amount of the lipid nanoparticle of claim 1.

20. The method of claim 19, further comprising administering to the patient an effective amount of an enzymatic cofactor, supplemental magnesium, supplemental zinc, or a combination of two or more of thereof.

21. The method of claim 20, wherein the enzymatic cofactor is oxidized nicotinamide adenine dinucleotide (NAD+), a precursor of oxidized nicotinamide adenine dinucleotide (NAD+), reduced nicotinamide adenine dinucleotide (NADH), or a combination thereof.

22. The method of claim 21, wherein the precursor of oxidized nicotinamide adenine dinucleotide is nicotinamide adenine dinucleotide, niacin, nicotinamide, nicotinamide riboside, dihydronicotinamide riboside, nicotinamide mononucleotide, dihydronicotinamide mononucleotide, tryptophan, or a combination of two or more thereof.

23. A nucleic acid encoding: (i) an aldehyde dehydrogenase 2 enzyme, wherein the nucleic acid has at least90% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:21; (ii) an alcohol dehydrogenase 1 enzyme, wherein the nucleic acid has at least90% sequence identity to SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6,SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24; or (iii) an aldehyde dehydrogenase 2 enzyme and an alcohol dehydrogenase 1enzyme, wherein the nucleic acid has at least 90% sequence identity to a nucleic acid comprising (i) SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:21, and (b) SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:

24.

24. An enzyme, wherein the enzyme is: (i) an aldehyde dehydrogenase 2 enzyme having an amino acid sequence withat least 90% sequence identity to SEQ ID NO:11 or SEQ ID NO:14, (ii) an alcohol dehydrogenase 1 enzyme having an amino acid sequence with atleast 90% sequence identity to SEQ ID NO:17 or SEQ ID NO:20, or (iii) an aldehyde dehydrogenase 2 enzyme having an amino acid sequence withat least 90% sequence identity to SEQ ID NO:11 or SEQ ID NO:14, and an alcohol dehydrogenase 1 enzyme having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:17 or SEQ ID NO:20.

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