NEW LIPIDS AND NANOPARTICLE COMPOSITIONS OF THESE.

Abstract

Lipids having Formula (I): (SEE FORMULA) and pharmaceutically acceptable salts thereof are provided herein, wherein R1, R2, a and b are as defined herein. Lipid nanoparticle (LNP) compositions comprising lipids having Formula (I) and a capsid-free, non-viral vector (e.g., eDNA) are also provided herein. In any aspect or embodiment hereof, these LNPs may be used to deliver a capsid-free, non-viral DNA vector to a target site of interest (e.g., cell, tissue, organ, and the like).

Description

NEW LIPIDS AND NANOPARTICLE COMPOSITIONS OF THESE RELATED REQUESTS This application claims the priority of United States provisional application no. 63 / 000,990, filed March 27, 2020, the entire contents of which are incorporated herein by reference. LIST OF SEQUENCES This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. Said ASCII copy, created on March 18, 2021, is called 131698-07720_SL.txt and has a size of 417 bytes. BACKGROUND Gene therapy aims to improve clinical outcomes for patients who have genetic disorders or acquired diseases caused by an abnormal gene expression profile. To date, several types of gene therapy have been developed that deliver therapeutic nucleic acids into a patient's cells as a drug to treat diseases. The delivery and expression of a corrective gene in the patient's target cells can be carried out by numerous methods, including the use of engineered viral gene delivery vectors and, potentially, plasmids, minigenes, oligonucleotides, minicircles or a variety of closed DNA. Among the many virus-derived vectors available (e.g., recombinant retrovirus, recombinant lenti vi rus, recombinant adenovirus, and the like), recombinant adeno-associated virus (rAAV) is gaining acceptance as a versatile and relatively reliable vector in gene therapy. However, viral vectors, such as adeno-associated vectors, can be highly immunogenic and elicit humoral and cell-mediated immunity that may compromise efficacy, particularly with respect to readministration. Nonviral gene delivery avoids certain disadvantages associated with viral transduction, particularly those due to humoral and cellular immune responses to the viral structural proteins that form the vector particle, and any de novo gene expression of the virus. Nonviral gene delivery technologies include the use of cationic lipids as a carrier. Ionizable lipids are approximately composed of an amine moiety and a lipid moiety, and a cationic amine moiety and a polyanionic nucleic acid interact electrostatically to form a positively charged liposome or lipid membrane structure. Therefore, uptake into cells is promoted and nucleic acids are delivered to cells. Some widely used ionizable lipids are CLinDMA, DLinDMA (also known as DODAP) and cationic lipids such as DOTAR. Of note, these lipids have been employed for siRNA delivery to the liver, but suffer from suboptimal delivery efficiency along with liver toxicity at higher doses. In view of the shortcomings of current cationic lipids, there is a need in the field to provide lipid structures that not only demonstrate improved efficacy along with reduced toxicity, but also improved pharmacokinetics and intracellular kinetics, such as cellular uptake. and the release of nucleic acid from the lipid carrier. SUMMARY In one aspect, provided herein are ionizable lipids having Formula (I): as well as pharmaceutically acceptable salts thereof, wherein R1, R2, a, and b are as defined herein. Further provided are pharmaceutical compositions comprising a described ionizable lipid, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier. Another aspect of the present description relates to a composition comprising a lipid nanoparticle (LNP) comprising an ionizable lipid described in the present description, or a pharmaceutically acceptable salt thereof, and a nucleic acid. In one embodiment of any of the aspects or embodiments of the present description, the nucleic acid is encapsulated in the ionizable lipid. In a particular embodiment, the nucleic acid is a closed DNA (ceDNA). According to some embodiments of any of the aspects or embodiments herein, the LNP further comprises an ester. According to some embodiments of any of the aspects or embodiments of the present description, the ester may be a cholesterol or betasitosterL. According to some embodiments of any of the aspects or embodiments herein, cholesterol is present in a molar percentage of about 20% to about 40%, for example about 20% to about 35%, about 20% to about 30%. , about 20% to about 25%, about 25% to about 35%, about 25% to about 30%, or about 30% to about 35%, and the ionizable lipid is present in a molar percentage of about 80% to about 35%, about 60%, for example about 80% to about 65%, about 80% to about 70%, about 80% to about 75%, about 75% to about 60%, about 75% to about 65%, about 75% to about 70%, about 70% to about 60%, or about 70% to about 60%. According to some embodiments of any of the aspects or embodiments herein, cholesterol is present in a molar percentage of about 20% to about 40%, for example about 20%, about 21%, about 22%, about 23%. , approximately 24%, approximately 25%, approximately 26%, approximately 27%, approximately 28%, approximately 29%, approximately 30%, approximately 31%, approximately 32%, approximately 33%, approximately 34%, approximately 35%, approximately 36%, about 37%, about 38%, about 39%, or about 40%, and wherein the ionizable lipid is present in a molar percentage of about 80% to about 60%, for example about 80%, about 79 %, approximately 78%, approximately 77%, approximately 76%, approximately 75%, approximately 74%, approximately 73%, approximately 72%, approximately 71%, approximately 70%, approximately 69%, approximately 68%, approximately 67% about 66%, about 65%, about 64%, about 63%, about 62%, about 61%, or about 60%. According to some embodiments of any of the aspects or embodiments herein, cholesterol is present at a molar percentage of approximately 40%, and wherein the ionizable lipid is present at a molar percentage of approximately 50%. According to some embodiments of any of the aspects or embodiments herein, the composition further comprises a cholesterol, a PEG-lipid conjugate and a non-cationic lipid. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate is present at about 1.5% to about 3%, for example about 1.5% to about 2.75%, about 1.5% to about 2.5%, about 1.5% to about 2.25%, about 1.5% to about 2%, about 2% to about 3%, about 2% to about 2.75%, about 2% to about 2.5%, about 2% to about 2.25%, about 2.25 % to about 3%, about 2.25% to about 2.75%, or about 2.25% to about 2.5%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate is present at about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, approximately 2%, approximately 2.1%, approximately 2.2%, approximately 2.3%, approximately 2.4%, approximately 2.5%, approximately 2.6%, approximately 2.7%, approximately 2.8%, approximately 2.9%, or approximately 3%. According to some embodiments of any of the aspects or embodiments herein, cholesterol is present in a molar percentage of about 30% to about 50%, for example about 30% to about 45%, about 30% to about 40%. , about 30% to about 35%, about 35% to about 50%, about 35% to about 45%, about 35% to about 40%, about 20% to about 40%, about 40% to about 50%, or about 45% to about 50%. According to some embodiments of any of the aspects or embodiments herein, cholesterol is present in a molar percentage of about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, approximately 37%, approximately 38%, approximately 39%, approximately 40%, approximately 41%, approximately 42%, approximately 43%, approximately 44%, approximately 45%, approximately 46%, approximately 47%, approximately 48% , approximately 49%, or approximately 50%. According to some embodiments of any of the aspects or embodiments herein, the LNP further comprises a polyethylene glycol (PEG)-lipid. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid is 1-(monomethoxypolyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG). According to some embodiments of any of the aspects or embodiments in the present description, the LNP further comprises a non-cationic lipid. According to some embodiments of any of the aspects or embodiments in the present description, the non-cationic lipid is selected from the group consisting of distearoyl-sn-glycero-phosphoethanolamine, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC) , dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPO), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoylphosphatidylethanolamine 4-(N-maleim¡domethyl)-cyclohexane-1-carboxylate (DOPE -mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), monomethyl-phosphatidylethanolamine (such as 16-O-monomethyl PE), dimethyl-phosphatidylethanolamine (such as 16-O-dimethyl PE), 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidiethanolamine (SOPE), hydrogenated soy phosphatidylcholine (HSPC), egg phosphatidylcholine (EPC), dioleoylphosphatidylserine (DOPS), sphingomyelin (SM), dimyristoyl phosphatidylcholine (DMPC) na (DLPE); 1,2-diphythanoyl-sn-glycero-3-phosphoethanolamine (DPHyPE); lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetylphosphate, lysophosphatidylcholine, dilinoleoylphosphatidylcholine or mixtures of these. According to some embodiments of any of the aspects or embodiments herein, the non-cationic lipid is selected from the group consisting of dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylcholine (DSPC) and dioleoylphosphatidylethanolamine (DOPE). According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate is present at about 1.5% to about 4%, for example about 1.5% to about 3%, about 2% to about 3%, about 2.5% to about 3%, about 1.5% to about 2.75%, about 1.5% to about 2.5%, about 1.5% to about 2.25%, about 1.5% to about 2%, about 1.5% to about 1.75%, about 2 % to about 3%, about 2% to about 2.75%, about 2% to about 2.5%, about 2% to about 2.25%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate is present at about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, about 2%, about 2.1%, approximately 2.2%, approximately 2.3%, approximately 2.4%, approximately 2.5%, approximately 2.6%, approximately 2.7%, approximately 2.8%, approximately 2.9%, or approximately 3%. According to some embodiments of any of the aspects or embodiments herein, the non-cationic lipid is present in a molar percentage of about 42.5% to about 62.5%. According to some embodiments of any of the aspects or embodiments herein, the ionizable lipid is present in a molar percentage of about 42.5%, about 43%, about 43.5%, about 44%, about 44.5%, about 45%. approximately 45.5%, approximately 46%, approximately 46.5%, approximately 47%, approximately 47.5%, approximately 48%, approximately 48.5%, approximately 49%, approximately 49.5%, approximately 50%, approximately 50.5%, approximately 51%, 51.5% , approximately 52%, approximately 52.5%, approximately 53%, approximately 53.5%, approximately 54%, approximately 54.5%, approximately 55%, approximately 55.5%, approximately 56%, approximately 56.5%, approximately 57%, 57.5%, approximately 58 %, approximately 58.5%, approximately 59%, approximately 59.5%, approximately M A / t / zuzz / uoy l OI 60%, approximately 60.5%, approximately 61%, approximately 61.5%, approximately 62%, or approximately 62.5%. According to some embodiments of any of the aspects or embodiments herein, the non-cationic lipid is present in a molar percentage of about 2.5% to about 12.5%. According to some embodiments of any of the aspects or embodiments herein, the cholesterol is present at a molar percentage of about 40%, the ionizable lipid is present at a molar percentage of about 52.5%, the non-cationic lipid is present at a molar percentage of approximately 7.5%, and where the PEG-lipid is present at approximately 3%. According to some embodiments of any of the aspects or embodiments herein, the LNP composition further comprises dexamethasone palmitate. According to some embodiments of any of the aspects or embodiments herein, the LNP has a size ranging between about 50 nm and about 110 nm in diameter, for example about 50 nm and about 100 nm, about 50 nm and about 95 nm. nm, approximately 50 nm and approximately 90 nm, approximately 50 nm and approximately 85 nm, approximately 50 nm and approximately 80 nm, approximately 50 nm and approximately 75 nm, approximately 50 nm and approximately 70 nm, approximately 50 nm and approximately 65 nm, about 50 nm and about 60 nm, about 50 nm and about 55 nm, about 60 nm and about 110 nm, about 60 nm and about 100 nm, about 60 nm and about 95 nm, about 60 nm and about 90 nm, about 60 nm and approximately 85 nm, approximately 60 nm and approximately 80 nm, approximately 60 nm and approximately 75 nm, approximately 60 nm and approximately 70 nm, approximately 60 nm and approximately 65 nm, approximately 70 nm and approximately 110 nm, approximately 70 nm and about 100 nm, about 70 nm and about 95 nm, about 70 nm and about 90 nm, about 70 nm and about 85 nm, about 70 nm and about 80 nm, about 70 nm and about 75 nm, about 80 nm and about 110 nm, approximately 80 nm and approximately 100 nm, approximately 80 nm and approximately 95 nm, approximately 80 nm and approximately 90 nm, approximately 80 nm and approximately 85 nm, approximately 90 nm and approximately 110 nm, or approximately 90 nm and approximately 100 nm . According to some embodiments of any of the aspects or embodiments herein, the LNP has a size less than about 100 nm, for example, a size less than about 105 nm, less than about 100 nm, less than about 95 nm, less than about 90 nm, less than about 90 nm, less than about 80 nm, less than about 75 nm, less than about 70 nm, less than about 65 nm, less than about 60 nm, less than about 55 nm, less than about 50 nm, less than about 45 nm, less than about 40 nm, less than about 35 nm, less than about 30 nm, less than about 25 nm, less than about 20 nm, less than about 15 nm, or less than about 10 nm. According to some embodiments of any of the aspects or embodiments herein, the LNP has a size of less than 70 nm, for example, less than 65 nm, less than 60 nm, less than about 55 nm, less than about 50 nm , less than about 45 nm, less than about 40 nm, less than about 35 nm, less than about 30 nm, less than about 25 nm, less than about 20 nm, less than about 15 nm, or less than about 10 nm. According to some embodiments, the LNP has a size of less than about 60 nm, for example, less than about 55 nm, less than about 50 nm, less than about 45 nm, less than about 40 nm, less than about 35 nm, less than about 30 nm, less than about 25 nm, less than about 20 nm, less than about 15 nm, or less than about 10 nm. According to some embodiments of any of the aspects or embodiments herein, the LNP composition has a total lipid to nucleic acid ratio of about 10:1. According to some embodiments of any of the aspects or embodiments herein, the LNP composition has a total lipid to nucleic acid ratio of about 20:1. According to some embodiments of any of the aspects or embodiments herein, the composition has a total lipid to nucleic acid ratio of about 30:1. According to some embodiments of any of the aspects or embodiments herein, the composition has a total lipid to nucleic acid ratio of about 40:1. According to some embodiments of any of the aspects or embodiments herein, the composition has a total lipid to nucleic acid ratio of about 50:1. According to some embodiments of any of the aspects or embodiments herein, the LNP further comprises a tissue targeting moiety. The tissue targeting portion may be a peptide, oligosaccharide or the like, which may be used for delivery of the LNP to one or more specific tissues, such as cancer, liver, CNS or muscle. According to some embodiments of any of the aspects or embodiments herein, the tissue targeting portion is attached to the PEG-lipid conjugate. According to some embodiments of any of the aspects or embodiments herein, the tissue targeting portion is a ligand for specific liver receptors. According to some embodiments of any of the aspects or embodiments herein, the liver-specific receptor ligand used for targeting to the liver is an oligosaccharide such as Nacetylgalactosamine (GalNAc). According to some embodiments of any of the aspects or embodiments herein, the GalNAc-linked PEG-lipid conjugate linked to GalNAc is present in the lipid nanoparticle in a molar percentage of 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate linked to GalNAc is present in the LNP at a molar percentage of 0.2%. According to some embodiments of any of the aspects or embodiments herein, the GalNAc-linked PEG-lipid conjugate is present in the LNP at a molar percentage of 0.3%. According to some embodiments of any of the aspects or embodiments herein, the GalNAc-linked PEG-lipid conjugate is present in the LNP at a molar percentage of 0.4%. According to some embodiments of any of the aspects or embodiments herein, the GalNAc-linked PEGlipid conjugate is present in the LNP at a molar percentage of 0.5%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate linked to GalNAc is present in the LNP at a molar percentage of 0.6%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate linked to GalNAc is present in the LNP at a molar percentage of 0.7%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate linked to GalNAc is present in the LNP at a molar percentage of 0.8%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate linked to GalNAc is present in the LNP at a molar percentage of 0.9%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate linked to GalNAc is present in the LNP at a molar percentage of 1.0%. According to some embodiments of any of the aspects or embodiments herein, the GalNAc-linked PEG-lipid conjugate is present in the LNP at a molar percentage of about 1.5%. According to some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate linked to GalNAc is present in the LNP at a molar percentage of 2.0%. According to some embodiments of any of the aspects or embodiments herein, the LNP composition is prepared in a buffer such as malic acid. According to some embodiments of any of the aspects or embodiments herein, the composition is prepared in about 10 mM to about 30 mM malic acid, for example about 10 mM to about 25 mM, about 10 mM to about 20 mM, about 10mM to about 15mM, about 15mM to about 25mM, about 15mM to about 20mM, about 20mM to about 25mM. According to some embodiments of any of the aspects or embodiments herein, the composition is prepared in about 10 mM melic acid, about 12 mM melic acid, about 14 mM melic acid, about 16 mM melic acid, about 18 mM melic acid, approximately 20 mM melic acid, approximately 22 mM melic acid, approximately 24 mM melic acid, approximately 26 mM melic acid, ma. t / zuzz / uoy 101 about 11 about 13 about 15 about 17 about 19 about 21 about 23 about 25 about 27 mM melic acid. mM melic acid, mM melic acid, mM melic acid mM melic acid, mM melic acid, mM melic acid. mM melic acid, mM melic acid, about 28 mM melic acid, about 29 mM melic acid, or about 30 mM melic acid. According to some embodiments of any of the aspects or embodiments herein, the composition comprises approximately 20 mM melic acid. According to some embodiments of any of the aspects or embodiments herein, the LNP composition is prepared in a solution having about 30 mM to about 50 mM NaCl, for example about 30 mM to about 45 mM NaCl, about 30 mM to about 40 mM NaCl, about 30 mM to about 35 mM NaCl, about 35 mM to about 45 mM NaCl, about 35 mM to about 40 mM NaCl, or about 40 mM to about 45 mM NaCl. According to some embodiments of any of the aspects or embodiments herein, the LNP composition is prepared in a solution having about 30 mM NaCl, about 35 mM NaCl, about 40 mM NaCl, or about 45 mM NaCI. According to some embodiments of any of the aspects or embodiments herein, the LNP composition is prepared in a solution having approximately 40 mM NaCl. According to some embodiments of any of the aspects or embodiments herein, the LNP composition is prepared in a solution having from about 20 mM to about 100 mM MgCls, for example about 20 mM to about 90 mM MgCl2, about 20 mM to about 80 mM MgCI2, about 20 mM to about 70 mM MgCI2, about 20 mM to about 60 mM MgCI2, about 20 mM to about 50 mM MgCI2, about 20 mM to about 40 mM MgCI2, about 20 mM to about 30 mM MgCl2, about 320 mM to about 90 mM MgCI2, about 30 mM to about 80 mM MgCh, about 30 mM to about 70 mM MgCI2, about 30 mM to about 60 mM MgCk, about 30mM to about 50mM MgCh, about 30mM to about 40mM MgCk, about 40mM to about 90mM MgCh, about 40mM to about 80mM MgCh, about 40mM to about 70mM MgCl, about 40mM to about 60mM MgCl, about 40mM to about 50mM MgCh, about 50mM to about 90mM MgCh, about 50mM to about 80mM MgCb, about 50mM to about 70mM MgCk, about 50mM to about 60mM MgCk, about 60mM to about 90mM MgCh, about 60mM to about 80mM MgCh, about 60mM to about 70mM MgCl, about 70mM to about 90mM MgCh, about 70 mM to about 80 mM MgCl, or about 80 mM to about 90 mM MgCl2. According to some embodiments of any of the aspects or embodiments herein, the ceDNA is closed-ended linear duplex DNA. According to some embodiments of any of the aspects or embodiments herein, the ceDNA comprises an expression cassette comprising a promoter sequence and a transgene. According to some embodiments of any of the aspects or embodiments herein, the ceDNA comprises an expression cassette comprising a polyadenylation sequence. According to some embodiments of any of the aspects or embodiments herein, the ceDNA comprises at least one inverted terminal repeat (ITR) flanking the 5' or 3' end of said expression cassette. According to some embodiments of any of the aspects or embodiments herein, the expression cassette is flanked by two ITRs, wherein the two ITRs comprise a 5' ITR and a 3' ITR. According to some embodiments of any of the aspects or embodiments herein, the expression cassette is connected to an ITR at the 3' end (3' ITR). According to some embodiments of any of the aspects or embodiments herein, the expression cassette is connected to an ITR at the 5' end (5' ITR). According to some embodiments of any of the aspects or embodiments herein, at least one of the 5' ITR and the 3' ITR is a wild-type AAV ITR. According to some embodiments of any of the aspects or embodiments herein, at least one of the 5' ITR and the 3' ITR is a modified ITR. According to some embodiments of any of the aspects or embodiments herein, the ceDNA further comprises a spacing sequence between a 5' ITR and the expression cassette. According to some embodiments of any of the aspects or embodiments herein, the ceDNA further comprises a spacing sequence between a 3' ITR and the expression cassette. According to some embodiments of any of the aspects or embodiments herein, ma / t / zuzz / uoy l OI the sword sequence has a length of at least 5 base pairs. According to some embodiments of any of the aspects or embodiments herein, the sword sequence has a length of 5 to 100 base pairs. According to some embodiments of any of the aspects or embodiments herein, the sword sequence has a length of 5,10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70 , 75, 80, 85, 90, 95 or 100 base pairs. According to some embodiments of any of the aspects or embodiments herein, the sword sequence has a length of 5 to 500 base pairs. According to some embodiments of any of the aspects or embodiments herein, the sword sequence is 5, 10,15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235,240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335,340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435,440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, or 495 base pairs in length. According to some embodiments of any of the aspects or embodiments herein, the ceDNA has a nick or a break. According to some embodiments of any of the aspects or embodiments herein, the ITR is an ITR derived from an AAV serotype, derived from a goose virus ITR, derived from a B19 virus ITR, a wild-type ITR from a parvovirus. According to some embodiments of any of the aspects or embodiments herein, the AAV serotype is selected from the group comprising AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11 and AAV12. According to some embodiments of any of the aspects or embodiments herein, the ITR is a mutant ITR, and the ceDNA optionally comprises an additional ITR that differs from the first ITR. According to some embodiments of any of the aspects or embodiments herein, the ceDNA comprises two mutant ITRs at the 5' and 3' ends of the expression cassette, where optionally the two mutant ITRs are symmetrical mutants. According to some embodiments of any of the aspects or embodiments herein, the ceDNA is a CELiD, a DNA-based minicircle, a MIDGE, a DNA ministrand, a dumbbell-shaped closed-end linear duplex DNA comprising two ITR hairpin structures at the 5' and 3' ends of an expression cassette, or a doggybone™ DNA. According to some embodiments of any of the aspects or embodiments herein, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient. According to some aspects, the disclosure provides a method of treating a genetic disorder in a subject, the method comprising administering to the subject an effective amount of the pharmaceutical composition according to any of the aspects or embodiments herein. In accordance with some modalities of any of the aspects or modalities hereof, MA / IZ / ¿U¿¿ / UO3 / ΟΊ the subject is a human being. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is selected from the group consisting of sickle cell anemia, melanoma, hemophilia A (coagulation factor VIII (FVIII) deficiency), and hemophilia B ( coagulation factor IX (FIX) deficiency, cystic fibrosis (CFTR), familial hypercholesterolemia (LDL receptor defect), hepatoblastoma, Wilson's disease, phenylketonuria (PKU), congenital hepatic porphyria, inherited disorders of hepatic metabolism, Lesch Nyhan, sickle cell anemia, thalassemias, xeroderma pigmentosum, Fanconi anemia, retinitis pigmentosa, ataxia telangiectasia, Bloom syndrome, retinoblastoma, mucopolysaccharide storage diseases (e.g., Hurler syndrome (MPS type I), Scheie syndrome (MPS type I S), Hurler-Scheie syndrome (MPS type I H-S), Hunter syndrome (MPS type II), Sanfilippo types A, B, C and D (MPS types III A, B, C and D), Morquio types A and B (MPS IVA and MPS IVB), Maroteaux-Lamy syndrome (MPS type VI), Sly syndrome (MPS type Vil), hyaluronidase defibrillation (MPS type IX)), Niemann-Pick disease types A / B, C1 and C2, Fabry disease, Schindler disease, GM2 gangliosidosis type II (Sandhoff disease), Tay-Sachs disease, metachromatic leukodystrophy, Krabbe disease, mucolipidosis types I, ll / lll and IV, sialidosis types I and II, glycogen storage disease types I and II (Pompe disease), Gaucher disease types I, II and III, Fabry disease, cystinosis, Batten disease, aspartylglucosaminuria, Salla disease, Danon disease (glycogen deficiency LAMP-2), lysosomal acid lipase (LAL) deficiency, neuronal ceroid lipofuscinosis (CLN1-8, INCL and LINCL), sphingolipidosis, galactosialidosis, amyotrophic lateral sclerosis (ALS), Parkinson's disease, Alzheimer's disease, Huntington's disease, spinocerebellar ataxia, spinal muscular atrophy, Friedreich's ataxia, Duchenne muscular dystrophy (DMD), Becker muscular dystrophies (BMD), dystrophic epidermolysis bullosa (DEB), ectonucleotide pyrophosphatase 1 deficiency, generalized arterial calcification of infancy (GACI), Leber congenital amaurosis, Stargardt macular dystrophy (ABCA4), ornithine transcarbamylase (OTC) deficiency, Usher syndrome, alpha-1 antitrypsin deficiency, progressive familial intrahepatic cholestasis (PFIC) type I (ATP8B1 deficiency), type II ( ABCB11), type III (ABCB4), or type IV (TJP2) and Cathepsin A deficiency. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is Leber congenital amaurosis (LOA). According to some embodiments of any of the aspects or modalities hereof, the LOA is LCA10. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is Niemann-Pick disease. According to some embodiments of any of the aspects or embodiments hereof, the genetic disorder is Niemann-Pick macular dystrophy. Stargardt. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is glucose-6-phosphatase (G6Pase) deficiency (glycogen storage disease type I) or Pompe disease (glycogen storage disease type I). MA / lz / ¿u¿¿ / uoy 101 glycogen storage type II). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is hemophilia A (factor VIII deficiency). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is hemophilia B (factor IX deficiency). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is Hunter syndrome (mucopolysaccharidosis II). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is cystic fibrosis. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is dystrophic epidermolysis bullosa (DEB). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is phenylketonuria (PKU). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is progressive familial intrahepatic cholestasis (PFIC). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is Wilson's disease. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is Gaucher disease type I, II or III. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is age-related macular degeneration. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is ornithine transcarbamylase deficiency. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is retinitis pigmentosa (RP1). According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is Usher syndrome. According to some embodiments of any of the aspects or embodiments herein, the genetic disorder is lysosomal acid lipase (LAL) deficiency. BRIEF DESCRIPTION OF THE DRAWINGS The embodiments of the present description, briefly summarized above and discussed in greater detail below, may be understood by reference to the illustrative embodiments of the description depicted in the accompanying drawings. However, the accompanying drawings illustrate only typical embodiments of the description and, therefore, should not be considered limiting of the scope, since the description may admit other equally effective embodiments. Figure 1 shows the improvements in luc expression in ceDNA achieved by employing the described lipid nanoparticles (e.g., LNP 5 comprising lipid 1 and LNP 6 comprising lipid 3) compared to SS-OP (e.g. , LNP 1,2 and 7-12) as observed in Study A. Figure 2 shows the improvements in luc expression in ceDNA achieved by employing the described lipid nanoparticles (e.g., LNP 16 comprising lipid 2, LNP 17 comprising lipid 1, and LNP 18 comprising lipid 3) in comparison with SS-OP (i.e., LNP 13), as seen in Study B. Figure 3 shows improvements in responsiveness at increased dosage levels, whereby an increased dose administered to mice leads to a further increase in luc expression in ceDNA, achieved by employing the lipid nanoparticles described ( for example, LNP 20 comprising lipid 1) compared to SS-OP (i.e., LNP 19), as observed in Study C. Figure 4A shows the improvements in luc expression in ceDNA achieved by employing the described lipid nanoparticles (e.g., LNP 24 comprising lipid 6, LNP 25 comprising lipid 7, and LNP 26 comprising lipid 8) in comparison with SS-OP (i.e., LNP 23), as observed in Study D. Figure 4B shows improvements in tolerability (as measured by change in body weight) in mice using the lipid nanoparticles described ( for example, LNP 24 comprising lipid 6, LNP 25 comprising lipid 7 and LNP 26 comprising lipid 8) compared to ionizable lipid A (i.e., LNP 22) used as a control. Figure 5A shows the improvements in luc expression in ceDNA achieved by employing the described lipid nanoparticles (e.g., LNP 28 comprising lipid 9 and LNP 29 comprising lipid 10) compared to SS-OP (i.e. , LNP 27). Figure 5B shows that improvements in luc expression in ceDNA as shown in Figure 5A did not compromise the tolerability in mice of the described lipid nanoparticles. DETAILED DESCRIPTION The present disclosure provides a lipid-based platform for delivering therapeutic nucleic acid (TNA) such as viral or non-viral vectors (e.g., closed-end DNA), which can move from the cytoplasm of the cell toward the nucleus, and maintain high expression levels. For example, the immunogenicity associated with viral vector-based gene therapies has limited the number of patients who can be treated due to pre-existing background immunity, as well as prevented redosing of patients, either to titrate to effective levels in each patient, or to maintain long-term effects. Furthermore, other nucleic acid modalities suffer greatly from immunogenicity due to an innate DNA or RNA sensing mechanism that triggers a cascade of immune responses. Due to the lack of pre-existing immunity, the currently described TNA lipid particles (e.g., lipid nanoparticles) allow additional doses of TNA, such as mRNA, siRNA, or ceDNA, as needed, and further expand patient access, including to pediatric populations who may require dosing after tissue growth. Furthermore, it is a finding of the present disclosure that TNA lipid particles (e.g., lipid nanoparticles), particularly comprising lipid compositions comprising one or more tertiary amino groups and a disulfide bond, provide more efficient delivery of TNA ( e.g. ceDNA), better tolerability and an improved safety profile. Because currently described TNA lipid particles (e.g., lipid nanoparticles) do not have packing restrictions imposed by space within the viral capsid, in theory, the only size limitation of TNA lipid particles (e.g., lipid nanoparticles) lies in the expression efficiency (e.g., DNA replication or RNA translation) of the host cell. One of the biggest obstacles in the development of therapeutic options, particularly in rare diseases, is the large number of specific conditions. About 350 million people worldwide live with rare disorders, defined by the National Institutes of Health as a disorder or condition with fewer than 200,000 people diagnosed. About 80 percent of these rare disorders are genetic in origin, and about 95 percent of them have no FDA-approved treatment (rarediseases.info.nih.gov / diseases / pages / 31 / faqs-about-rare- diseases). Among the advantages of the TNA lipid particles (e.g., lipid nanoparticles) described herein is providing a method that can be easily adapted to multiple diseases that can be treated with a specific TNA modality, and in particular to rare monogenic diseases. that can significantly change the current state of treatments for many genetic disorders or diseases. L DEFINITIONS The term alkyl refers to a straight-chain (i.e., unbranched) or branched monovalent saturated hydrocarbon radical. Illustrative alkyl groups include, but are not limited to, ethyl, propyl, isopropyl, 2-methyl-1-butyl, 3-methyl-2-butyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4 -methyl1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2- ethyl-1-butyl, butyl, sobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decanyl, undecanyl, dodecanyl, tridecanyl, tetradecanyl, pentadecanyl, hexadecanyl, heptadecanyl, octadecanyl, nonadecanyl, eicosanyl , etc. The term alkenyl refers to a linear or branched aliphatic hydrocarbon radical with one or more (e.g., one or two) carbon-carbon double bonds, wherein the alkenyl radical includes radicals having "cis" and "trans" orientations, or by alternative nomenclature, guidelines The term pharmaceutically acceptable salt, as used herein, refers to pharmaceutically acceptable organic or inorganic salts of an ionizable lipid of the invention. Examples of salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate , bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate mesylate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate (i.e., 1,1'-methylene-bis- (2-hydroxy¡3-naphthoate)), alkali metal salts (for example, sodium and potassium), alkaline earth metal salts (for example, magnesium) and ammonium salts. A pharmaceutically acceptable salt may involve the inclusion of another molecule such as an acetate ion, a succinate ion or another counterion. The counterion can be any organic or inorganic moiety that stabilizes the charge in the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. Cases where multiple charged atoms are part of the pharmaceutically acceptable salt may have multiple counterions. Therefore, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions. As used in this description and the accompanying claims, the term approximately, when referring to a measurable value such as a quantity, a temporal duration and the like, is intended to encompass variations of ±20% or ±10%, more preferably, ±5%, even more preferably ±1% and even more preferably ±0.1% of the specified value, as such variations are appropriate for performing the methods described. As used herein, understand, comprising and comprising and composed of are synonyms for include, including, includes or contain, containing, contains and are inclusive or open terms that specify the presence of what follows, e.g. , a component, and do not exclude or prevent the presence of additional components, features, elements, members or steps not mentioned, known in the art or described therein. The term consisting of refers to compositions, methods, processes, and their respective components as described herein, which are exclusive of any element not mentioned in that description of the embodiment. As used herein, the term essentially consisting of refers to those elements required for a given embodiment. The expression allows the presence of additional elements that do not materially affect the basic and novel or functional characteristics of that modality of the invention. As used herein, the terms administration, administer and variants thereof refer to the introduction of a composition or agent (e.g., nucleic acids, in particular ceDNA) into a subject and includes the simultaneous and sequential introduction of one or more compositions or agents. Administration may refer, for example, to therapeutic, pharmacokinetic, diagnostic, investigational, placebo and experimental methods. Administration also encompasses in vitro and ex vivo treatments. The introduction of a composition or agent into a subject is by any suitable route, including oral, pulmonary, intranasal, parenteral (intravenous, intramuscular, intraperitoneal or subcutaneous), rectal, intralymphatic, intratumoral or topical. Administration includes self-administration and administration by another person. Administration can be carried out by any suitable route. A suitable route of administration allows the composition or agent to perform the desired function. For example, if a suitable route is intravenous, the composition is administered by introducing the composition or agent into a vein of the subject. In one aspect of any of the aspects or embodiments herein, administration refers to therapeutic administration. As used herein, the phrase immune response against therapeutic nucleic acid, immune response against transfer vector, immune response against a therapeutic nucleic acid, immune response against a transfer vector or the like, refers to any response unwanted immune response to a therapeutic nucleic acid, viral or non-viral in origin. In some embodiments of any of the aspects and embodiments herein, the unwanted immune response is an antigen-specific immune response against the viral transfer vector itself. In some embodiments of any of the aspects or embodiments herein, the immune response is specific to the transfer vector which may be double-stranded DNA, single-stranded RNA, or double-stranded RNA. In other embodiments, the immune response is specific to a sequence of the transfer vector. In other embodiments, the immune response is specific to the CpG content of the transfer vector. As used herein, the terms carrier and excipient are intended to include any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic agents and absorption retarding agents, buffers, carrier solutions, suspensions, colloids and the like. The use of such means and agents for pharmaceutically active substances is known in the art. In addition, complementary active ingredients can be incorporated into the compositions. The phrase pharmaceutically acceptable refers to molecular entities and compositions that do not produce a toxic or allergic reaction, or other similar inappropriate reaction, when administered to a host. As used herein, the term ceDNA is intended to refer to capsid-free linear closed-end duplex (be) DNA for non-viral, synthetic or other gene transfer. The detailed description of ceDNA is described in international application PCT / US2017 / 020828, filed on March 3, 2017, the entire contents of which are expressly incorporated herein by reference. Certain methods for the production of ceDNA that ΜΛ / t / zuzz / uoy l OI comprise various inverted terminal repeat (ITR) sequences and configurations using cell methods are described in Example 1 of international applications PCT / US18 / 49996, filed on September 7, 2018, and PCT / US2018 / 064242, filed December 6, 2018, each of which is incorporated herein by reference in its entirety. Certain methods for the production of synthetic ceDNA vectors comprising various ITR sequences and configurations are described, for example, in international application PCT / US2019 / 14122, filed on January 18, 2019, the full contents of which are incorporated herein. as reference. As used herein, the terms ceDNA vector and ceDNA are used interchangeably. According to some embodiments of any of the aspects or embodiments herein, the ceDNA is closed-end linear duplex DNA (CELiD) CELiD DNA. According to some embodiments of any of the aspects or embodiments herein, the ceDNA is a DNA-based minicircle. According to some embodiments of any of the aspects or embodiments herein, ceDNA is an immunologically defined minimalist expression (MIDGE) vector. According to some embodiments of any of the aspects or embodiments herein, the ceDNA is a ministrand DNA. According to some embodiments of any of the aspects or embodiments herein, the ceDNA is dumbbell-shaped linear closed-end duplex DNA comprising two ITR hairpin structures at the 5' and 3' ends of a cassette. expression. According to some embodiments of any of the aspects or embodiments herein, the ceDNA is a doggybone™ DNA. As used herein, the term ceDNA bacmid is intended to refer to an infectious baculovirus genome that comprises a ceDNA genome as an intermolecular duplex that is capable of propagating in E. coli as a plasmid, and thus can function as a vector. shuttle for baculovirus. As used herein, the term ceDNA baculovirus is intended to refer to a baculovirus that comprises a ceDNA genome as an intermolecular duplex within the baculovirus genome. As used herein, the terms ceDNA baculovirus-infected insect cell and ceDNA-BIIC are used interchangeably and are intended to refer to an invertebrate host cell (including, but not limited to, an insect cell (e.g. an Sf9)) cell infected with a ceDNA baculovirus. As used herein, the term ceDNA genome is intended to refer to an expression cassette that further incorporates at least one inverted terminal repeat region. A ceDNA genome may further comprise one or more spacing regions. In some embodiments of any of the aspects or embodiments herein, the ceDNA genome is incorporated as an intermolecular DNA duplex polynucleotide into a plasmid or viral genome. a promoter or other DNA regulatory sequence sufficient to direct transcription of a transgene from a DNA vector, e.g., synthetic AAV vector. Suitable promoters include, for example, tissue-specific promoters. Promoters may also be of AAV origin. As used herein, the term flanking is intended to refer to a relative position of one nucleic acid sequence in relation to another nucleic acid sequence. Generally, in the ABC sequence, B is flanked by A and C. The same is true in the AxBxC arrangement. Therefore, a flanking sequence precedes or follows a flanked sequence, but need not necessarily be contiguous or immediately adjacent to the flanked sequence. In one embodiment of any of the aspects or embodiments herein, the term flanking refers to terminal repeats at each end of the linear single-stranded synthetic AAV vector. As used herein, the term gene is broadly used to refer to any segment of nucleic acid associated with the expression of a given RNA or protein, in vitro or in vivo. Genes therefore include regions that encode expressed RNAs (typically including sequences that encode polypeptides) and often the regulatory sequences necessary for their expression. Genes can be obtained from a variety of sources, including cloning from a source of interest or synthesis from known or predicted sequence information, and can include sequences designed to have specifically desired parameters. As used herein, the phrase genetic disease or genetic disorder is intended to refer to a disease caused, partially or completely, directly or indirectly, by one or more abnormalities in the genome, including, and especially, a condition that has been present since the birth. The abnormality may be a mutation, insertion, or deletion in a gene. The abnormality may affect the coding sequence of the gene or its regulatory sequence. The term heterologous, as used herein, is intended to refer to a nucleotide or polypeptide sequence that is not found in the native nucleic acid or protein, respectively. A heterologous nucleic acid sequence can be ligated to a naturally occurring nucleic acid sequence (or a variant thereof) (e.g., by genetic engineering) to generate a chimeric nucleotide sequence encoding a chimeric polypeptide. A heterologous nucleic acid sequence can be ligated to a variant polypeptide (e.g., by genetic engineering) to generate a nucleotide sequence encoding a fusion variant polypeptide. As used herein, the term host cell refers to any type of cell that is susceptible to transformation, transfection, transduction and the like with nucleic acid therapeutic options of the present description. As non-limiting examples, a host cell may be an isolated primary cell, pluripotent stem cells, CD34+ cells, induced pluripotent stem cells, or any of a number of immortalized cell lines (e.g., MA / IZ / ¿U¿¿ / UO3 / 01 HepG2 cells). Alternatively, a host cell may be a cell in situ or in vivo in a tissue, organ or organism. Furthermore, a host cell may be a target cell, for example, of a mammalian subject (for example, a human patient in need of gene therapy). As used herein, an inductive promoter is intended to refer to one that is characterized by initiating or enhancing transcriptional activity when in the presence of, influenced by, or in contact with an inducer or inducing agent. An inducer or inducing agent, as used herein, may be endogenous, or a normally exogenous compound or protein that is administered in such a way that it is active to induce the transcriptional activity of the inducible promoter. In some embodiments of any of the aspects or embodiments herein, the inducer or inducing agent, that is, a chemical substance, a compound or a protein, may itself be the result of the transcription or expression of a nucleic acid sequence. (i.e., an inducer may be an inducer protein expressed by another component or module), which may itself be under the control of an inducible promoter. In some embodiments of any of the aspects or embodiments herein, an inducible promoter is induced in the absence of certain agents, such as a repressor. Examples of inductive promoters include, but are not limited to, tetracycline, metallothionine, ecdysone, mammalian viruses (e.g., the adenovirus late promoter; and the mouse mammary tumor virus long terminal repeat (MMTV-LTR)), and other spheroid-responsive promoters, rapamycin-responsive promoters and the like. As used herein, the term in vitro is intended to refer to assays and methods that do not require the presence of a cell with an intact membrane, such as cell extracts, and may refer to the introduction of a programmed synthetic biological circuit into a system. non-cellular, such as a medium that does not comprise cells or cellular systems, such as cell extracts. As used herein, the term in vivo is intended to refer to assays or processes that occur in or within an organism, such as a multicellular animal. In some of the aspects described herein, a method or use may be said to occur in vivo when a single-celled organism, such as a dacteria, is used. The term ex vivo refers to methods and uses that are carried out with a living cell with an unaltered membrane that is outside the organism of a multicellular animal or plant, for example, explants, cultured cells, including primary cells and cell lines, cell lines. transformed cells and extracted tissue or cells, which include blood cells, among others. As used herein, the term lipid is intended to refer to a group of organic compounds that includes, but is not limited to, esters of fatty acids and are characterized by not being hydrosoluble, but soluble in many organic solvents. They are generally divided into at least three classes: (1) simple lipids, which include fats and oils as well as waxes; (2) lipids MA / t / zuzz / uoy / 01 compounds, including phospholipids and glycolipids; and (3) derived lipids such as steroids. Representative examples of phospholipids include, but are not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoyl phosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, distearoylphosphatidylcholine, and dilinoleoylphosphatidylcholine. hill. Other compounds that lack phosphorus, such as sphingolipids, glycosphingolipid families, diacylglycerols and β-acyloxy acids, are also found within the group designated as unsympathetic lipids. Additionally, the unsympathetic lipids described above can mix with other lipids, including triglycerides and sterols. In one embodiment of any of the aspects or embodiments herein, the lipid compositions comprise one or more tertiary amino groups, one or more phenyl ester bonds and a disulfide bond. As used herein, the term lipid conjugate is intended to refer to a lipid conjugate that inhibits the aggregation of lipid particles (e.g., lipid nanoparticles). Such lipid conjugates include, but are not limited to, PEG-lipid conjugates such as, for example, PEG coupled to dialkyloxypropyls (e.g., PEG-DAA conjugates), PEG coupled to diacylglycerols (e.g., PEG-DAG conjugates ), PEG coupled to cholesterol, PEG coupled to phosphatidylethanolamines, and PEG conjugated to ceramides (see, for example, US Patent No. 5,885,613), ionizable PEG lipids, polyoxazoline (POZ)lipid conjugates (e.g., POZ-DAA conjugates; see, for example, US Provisional Application No. 61 / 294,828, filed January 13, 2010 and US Provisional Application No. 61 / 295,140, ​​filed January 14, 2010 ), polyamide oligomers (e.g., ATTA-lipid conjugates) and mixtures thereof. In PCT publication no. Additional examples of POZ-lipid conjugates are described in WO 2010 / 006282. PEG or POZ can be conjugated directly to the lipid or can be attached to the lipid through a linker moiety. Any suitable linker moiety can be used to couple PEG or POZ to a lipid, including, for example, non-ester containing linker moieties and ester containing linker moieties. In certain preferred embodiments, non-ester containing linking moieties, such as amides or carbamates, are used. The descriptions of each of the foregoing patent documents are incorporated herein by reference in their entirety for all purposes. A lipid conjugate described herein (for example, PEG-lipid or PEGylated lipid can be covalently linked to a useful tissue targeting moiety known in the art (for example, N-acetylgalactosamine (GalNAc; GalNAc mono, di, tri or tetraantennary). As used herein, the term "lipid-encapsulated" is intended to refer to a lipid particle that provides an active agent or therapeutic agent, such as a nucleic acid (e.g., an ASO, mRNA, siRNA, ceDNA, viral vector), with complete encapsulation, partial iviA / t / zuzz / uoy roí encapsulation or both. In a preferred embodiment, the nucleic acid is completely encapsulated in the lipid particle (for example, to form a nucleic acid-containing lipid particle). As used herein, the terms lipid particle or lipid nanoparticle are intended to refer to a lipid formulation that can be used to deliver a therapeutic agent such as therapeutic nucleic acid agents (TNA) to a target site of interest (e.g., cell, tissue, organ and the like) (called TNA lipid particles, TNA lipid nanoparticles or TNA LNPs). In one embodiment of any of the aspects or embodiments herein, the lipid particle of the invention is a lipid particle containing therapeutic nucleic acid, which is typically formed from an ionizable lipid, a non-cationic lipid and, optionally, a conjugated lipid that prevents particle aggregation. In other preferred embodiments, a therapeutic agent such as a therapeutic nucleic acid can be encapsulated in the lipid portion of the particle, thereby protecting it from enzymatic degradation. In one embodiment of any of the aspects or embodiments herein, the lipid particle comprises a nucleic acid (e.g., ceDNA) and a lipid comprising one or more tertiary amino groups, one or more phenyl ester bonds and a disulfide bond. The lipid particles of the invention typically have a mean diameter of about 20 nm to about 120 nm, about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, about 70 nm to about 80 nm, or about 30 nm, about 35 nm, about 40 nm, about 45 nm, about 50 nm, about 55 nm, about 60 nm, about 65 nm, about 70nm, about 75nm, about 80nm, about 85nm, about 90nm, about 95nm, about 100nm, about 105nm, about 110nm, about 115nm, about 120nm, about 125nm, about 130nm , approximately 135 nm, approximately 140 nm, approximately 145 nm, or approximately 150 nm. As used herein, the term hydrophobic lipid refers to compounds having nonpolar groups including, but not limited to, long chain saturated and unsaturated aliphatic hydrocarbon groups and such groups optionally substituted by one or more aromatic, cycloaliphatic groups. or heterocyclic. Suitable examples include, but are not limited to, Μλ. t / zuzz / uoy 101 diacylglycerol, dialkylglycerol, N-N-dialkylamino, 1,2-diacyloxy¡-3-aminopropane and 1,2-dialkyl-3aminopropane. As used herein, the term ionizable lipid is intended to refer to a lipid, for example, a cationic lipid, that has at least one protonable or deprotonable group, such that the lipid is positively charged at a pH equal to or lower than physiological pH (for example, pH 7.4), and neutral at a second pH, preferably equal to or higher than the physiological pH. One skilled in the art will understand that the addition or removal of protons as a function of pH is an equilibrium process, and that reference to a charged or neutral lipid refers to the nature of the predominant species and does not require that the entire lipid be present in the charged or neutral form. Generally, ionizable lipids have a pKa of the protonable group in the range of about 4 to about 7. In some embodiments of any of the aspects and embodiments herein, an ionizable lipid may include cleavage lipid or SS cleavage lipid. Accordingly, the term ionizable lipid, as used herein, encompasses both ionized (or charged) and neutral forms of the lipids of the invention. As used herein, the term neutral lipid is intended to refer to any of the lipid species that exist in the form of an uncharged or neutral dipole ion at a selected pH. At physiological pH, such lipids include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides and diacylglycerols. As used herein, the term anionic lipid refers to any lipid that has a negative charge at physiological pH. These lipids include, but are not limited to, phosphatidylglycerols, cardiolipins, diacylphosphatidylserines, diacylphosphatidic acids, N-dodecanoyl phosphatidylethanolamines, Nsuccinyl phosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleiolphosphatidylglycerol (POPG), and other anionic modifying groups attached to neutral lipids. As used herein, the term non-cationic lipid is intended to refer to any antipathic lipid, as well as any other neutral lipid or anionic lipid. As used herein, the term cleavage lipid or SS cleavage lipid refers to a lipid comprising a disulfide bond cleavage unit. In one embodiment of any of the aspects and embodiments herein, the cleavable lipids comprise a tertiary amine, which responds to an acidic compartment, for example, an endosome or lysosome for membrane destabilization and a disulfide bond that can be cleaved into a reducing environment, such as the cytoplasm. In one embodiment of any of the aspects and embodiments herein, a cleavable lipid is an ionizable lipid. In one embodiment of any of the aspects and embodiments herein, a cleavable lipid is a cationic lipid. In one embodiment of any of the aspects and embodiments herein, a cleaved lipid is an ionized cationic lipid. Cleavable lipids are described in more detail herein. MA / IZ / ¿U¿¿ / UO3101 As used herein, the term organic lipid solution is intended to refer to a composition comprising, in whole or in part, an organic solvent having a lipid. As used herein, the term liposome refers to lipid molecules assembled in a spherical configuration that encapsulates an inner aqueous volume that is secreted from an aqueous exterior. Liposomes are vesicles that have at least one lipid bilayer. Liposomes are typically used as carriers for therapeutic agent / drug delivery in the context of pharmaceutical development. These work by fusing with a cell membrane and repositioning its lipid structure to deliver a drug or active pharmaceutical ingredient. Liposome compositions for such administration are typically composed of phospholipids, especially compounds having a phosphatidylcholine group, however, these compositions may also include other lipids. As used herein, the term local delivery refers to the delivery of an active agent such as an interfering RNA (e.g., siRNA) directly to a target site within an organism. For example, an agent can be delivered locally by direct injection into a diseased area such as a tumor or another target site such as a site of inflammation or a target organ such as the liver, heart, pancreas, kidney and the like. As used herein, the term neDNA or nicked ceDNA is intended to refer to a closed-end DNA that has a nick or gap of 2-100 base pairs in a stem region or sword region 5' upstream of a open reading frame (for example, a promoter and a transgene to be expressed). As used herein, the term nucleic acid is intended to refer to a polymer containing at least two nucleotides (i.e., deoxyribonucleotides or ribonucleotides) in single-stranded or double-stranded form and includes DNA, RNA, and hybrids thereof. The DNA may be in the form of, for example, antisense molecules, plasmid DNA, DNA-DNA duplexes, pre-condensed DNA, PCR products, vectors (P1, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA or derivatives and combinations of these groups. The DNA can be in the form of a minicircle, plasmid, bacmid, minigene, ministrand DNA (covalently closed linear DNA vector), closed-end linear duplex DNA (CELID or ceDNA), doggybone™ DNA, dumbbell DNA, minimalist immunologically defined gene expression vector (MIDGE), viral vector or non-viral vectors. The RNA may be in the form of a small interfering RNA (siRNA), Dicer substrate dsRNA, small hairpin RNA (hpRNA), asymmetric interfering RNA (iaRNA), microRNA (miRNA), mRNA, rRNA, tRNA, viral RNA (vRNA) and combinations of these. Nucleic acids include nucleic acids containing analogues of known nucleotides or modified backbone residues or linkages, which are synthetic, naturally occurring and non-naturally occurring, and which have binding properties similar to those of the reference nucleic acid. Examples of such analogues and / or modified residues include, without limitation, phosphorothioates, morpholino phosphorodiamidate oligomer (morpholino), phosphoramidates, methylphosphonates, chiralmethylphosphonates, 2'-0-methyl ribonucleotides, locked nucleic acid (LNA™) and peptide nucleic acids. (PNA). Unless specifically limited, the term encompasses nucleic acids containing known analogues of naturally occurring nucleotides that have binding properties similar to those of the reference nucleic acid. Unless otherwise indicated, a particular nucleic acid sequence further implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences, as well as the explicitly stated sequence. . As used herein, the phrases nucleic acid therapeutic agents, therapeutic nucleic acid, and TNA are used interchangeably and refer to any embodiment of therapeutic agent that uses nucleic acids as an active therapeutic agent component to treat a disease or disorder. As used herein, these phrases refer to RNA-based therapeutic agents and DNA-based therapeutic agents. Non-limiting examples of RNA-based therapeutic agents include mRNA, antisense RNA and oligonucleotides, ribozymes, aptamers, RNA interference (RNAi), Dicer substrate dsRNA, small hairpin RNA (shRNA), asymmetric RNA interference (aRNA). , and microRNA (miRNA). Non-limiting examples of DNA-based therapeutic agents include minicircle DNA, minigene, viral DNA (e.g., AAV or lentivirus genome) or non-viral DNA vectors, closed-end linear duplex DNA (ceDNA / CELID), plasmids, bacmids, doggybone™ DNA vectors, minimalist immunologically defined gene expression (MIDGE) vector, non-viral ministrand DNA vector (covalently closed linear DNA vector) and minimal dumbbell DNA vector ( dumbbell). As used herein, the term LNP with TNA refers to a lipid particle that contains at least one of the TNAs described above. As used herein, nucleotides contain a deoxyribose (DNA) or ribose (RNA) sugar, a base, and a phosphate group. The nucleotides are linked together through phosphate groups. As used herein, operatively joined refers to a juxtaposition where the components so described are in a relationship that allows them to function in the intended manner. For example, a promoter is operably linked to a coding sequence if the promoter affects its transcription or expression. A promoter can be said to direct the expression or transcription of the nucleic acid sequence it regulates. The phrases operatively linked, operatively located, under control, and under transcriptional control indicate that a promoter is in a correct functional location and / or orientation with respect to a nucleic acid sequence that it regulates to control the initiation and / or transcriptional expression of that sequence. . An inverted promoter, as used herein, refers to a promoter in which the sequence of ΜΛ / t / zuzz / uoy 101 nucleic acids is in the reverse orientation, so that what was the coding strand is now the non-coding strand and vice versa. Inverted promoter sequences can be used in various embodiments to regulate the state of a switch. Furthermore, in various embodiments, a promoter may be used together with an enhancer. As used herein, the term promoter is intended to refer to any nucleic acid sequence that regulates the expression of another nucleic acid sequence by driving transcription of the nucleic acid sequence, which may be a heterologous target gene that encodes a protein or RNA. Promoters can be constitutive, inductive, repressive, tissue specific or any combination of these. A promoter is a control region of a nucleic acid sequence in which the initiation and rate of transcription of the remainder of a nucleic acid sequence are controlled. A promoter may also contain genetic elements to which proteins and regulatory molecules, such as RNA polymerase and other transcription factors, can bind. Within the promoter sequence will be a transcription start site as well as protein binding domains responsible for RNA polymerase binding. Eukaryotic promoters often, but not always, contain TATA sequences and CAT sequences. Various promoters, including inducible promoters, can be used to drive expression of transgenes in the synthetic AAV vectors described herein. A promoter sequence may be bounded at its 3' terminus by the transcription start site and extends upstream (direction 5j to include the minimum number of bases or elements necessary to initiate transcription at detectable levels above background. A promoter may be one naturally associated with a gene or sequence, as may be obtained by isolating the 5' non-coding sequences located upstream of the coding segment and / or exon of a given gene or sequence. Such a promoter can be called endogenous. Similarly, in some embodiments of any of the aspects and embodiments herein, an enhancer may be one naturally associated with a nucleic acid sequence, located downstream or upstream of that sequence. In some embodiments of any of the aspects and embodiments herein, a coding nucleic acid segment is placed under the control of a recombinant promoter or heterologous promoter, both of which refer to a promoter that is not normally associated with the acid sequence. encoded nucleic acid to which it is operatively linked in its natural environment. Similarly, a recombinant or heterologous enhancer refers to an enhancer that is not normally associated with a given nucleic acid sequence in its natural environment. Such promoters or enhancers may include promoters or enhancers of other genes; promoters or enhancers isolated from any other prokaryotic, viral or eukaryotic cell; and synthetic promoters or enhancers that are not natural, that is, they comprise elements MA / lz / ¿u¿¿ / uoy 101 different transcription regulatory regions and / or mutations that alter expression through genetic engineering methods known in the art. In addition to producing promoter and enhancer nucleic acid sequences synthetically, promoter sequences can be produced by recombinant cloning technology and / or nucleic acid amplification, including POR, in connection with the synthetic biological circuits and modules described herein (see , for example, United States Patent No. 4,683,202 and United States Patent No. 5,928,906, each incorporated herein by reference in its entirety). Furthermore, it is contemplated that control sequences that direct the transcription and / or expression of sequences within non-nuclear organelles, such as mitochondria, chloroplasts, and the like, may also be used. As used herein, the terms Rep binding site (RBS) and Rep binding element (RBE) are used interchangeably and are intended to refer to a binding site for Rep protein (e.g., Rep 78 of AAV or Rep 68 of AAV) which, upon binding by a Rep protein, allows the Rep protein to perform its site-specific endonuclease activity on the sequence incorporating the RBS. An RBS sequence and its inverse complement together form a single RBS. RBS sequences are well known in the art and these include, for example, 5'GCGCGCTCGCTCGCTC-3', an RBS sequence identified in AAV2. As used herein, the phrase "recombinant vector" is intended to refer to a vector that includes a heterologous nucleic acid sequence or transgene that can be expressed in vivo. It will be understood that the vectors described herein may, in some embodiments of any of the aspects and embodiments herein, be combined with other suitable compositions and therapies. In some embodiments of any of the aspects and embodiments herein, the vector is episomal. The use of a suitable episomal vector provides a means of maintaining the nucleotide of interest in the subject in extra high copy number chromosomal DNA, thus eliminating the possible effects of chromosomal integration. As used herein, the term indicator refers to a protein that can be used to provide a detectable readout. An indicator generally produces a measurable signal such as fluorescence, color, or luminescence. Reporter protein coding sequences encode proteins whose presence in the cell or organism is easily observed. As used herein, the terms sense and antisense refer to the orientation of the structural element in the polynucleotide. The sense and antisense versions of one element are the inverse complement of the other. As used herein, the term sequence identity refers to the similarity between two nucleotide sequences. For the purposes of the present description, the degree of sequence identity between two deoxyribonucleotide sequences is determined by the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, supra) as implemented in the program Needle from the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, supra), preferably version 3.0.0 or later. The optional parameters used are the break open penalty of 10, the break extension penalty of 0.5, and the substitution matrix EDNAFULL (NCBI NUC4.4 EMBOSS version). The Needle result marked as longest identity (obtained using the -nobrief option) is used as the percent identity and is calculated as follows: (Identical deoxyribonucleotides.times.100) / (Alignment length-Total number of interruptions in the alignment). The alignment length is preferably at least 10 nucleotides, preferably at least 25 nucleotides, more preferably at least 50 nucleotides, and most preferably at least 100 nucleotides. As used herein, the term sword region is intended to refer to an intermediate sequence that separates functional elements in the vector or genome. In some embodiments of any of the aspects and embodiments herein, the AAV spacing regions maintain two functional elements at a desired distance for optimal functionality. In some embodiments of any of the aspects and embodiments herein, the spacing regions provide or add genetic stability to the vector or genome. In some embodiments of any of the aspects and embodiments herein, the spacing regions facilitate easy genetic manipulation of the genome by providing a convenient location for cloning sites and a base pair design number spacing. For example, in certain aspects, a polylinker or oligonucleotide polycloning site containing multiple restriction endonuclease sites, or a non-open reading frame sequence designed to have no known protein binding sites (e.g., transcription factor ), can be located in the vector or genome to separate the factors that act in ois, for example, by inserting a 6mer, 12mer, 18mer, 24mer, 48mer, 86mer, 176mer, etc. The term subject, as used herein, is intended to refer to a human or animal, to whom treatment, including prophylactic treatment, is provided with the therapeutic nucleic acid in accordance with the present invention. Generally, the animal is a vertebrate such as, but not limited to, a primate, rodent, domestic animal, or game animal. Primates include, but are not limited to, chimpanzees, cynomolgus monkeys, spider monkeys and macaques, e.g. Rhesus. Rodents include mice, rats, groundhogs, ferrets, rabbits and hamsters. Domestic and game animals include, but are not limited to, cows, horses, pigs, deer, bison, buffalo, feline species, for example, domestic cat, canine species, for example, dog, fox, wolf, bird species, for example, chicken, emu, ostrich, and fish, for example, trout, catfish and salmon. In certain embodiments of the aspects described herein, the subject is a mammal, for example, a primate or a human. A subject can be male or female. Additionally, a subject can be an infant or child. In some modalities of any of the MA / lz / ¿u¿¿ / uoy / ΟΊ aspects and modalities of the present, the subject may be a newborn or unborn subject, for example, the subject is in the womb. Preferably, the subject is a mammal. The mammal may be a human, non-human primate, mouse, rat, dog, cat, horse or cow, but is not limited to these examples. Non-human mammals can be used advantageously as subjects representing animal models of diseases and disorders. Additionally, the methods and compositions described herein can be used for domestic animals and / or pets. A human subject can be of any age, gender, race or ethnic group, for example, Caucasian (white), Asian, African, Black, African American, African European, Hispanic, Middle Eastern, etc. In some embodiments of any of the aspects and modalities herein, the subject may be a patient or another subject in a clinical setting. In some embodiments of any of the aspects and modalities herein, the subject is already under treatment. In some embodiments of any of the aspects and embodiments herein, the subject is an embryo, a fetus, a newborn, an infant, a child, an adolescent, or an adult. In some embodiments of any of the aspects and embodiments herein, the subject is a human fetus, a human newborn, a human infant, a human child, a human adolescent, or a human adult. In some embodiments of any of the aspects and embodiments herein, the subject is an animal embryo, or a non-human embryo, or a non-human primate embryo. In some embodiments of any of the aspects and embodiments herein, the subject is a human embryo. As used herein, the phrase "subject in need thereof" refers to a subject to whom (i) a TNA lipid particle (or a pharmaceutical composition comprising a TNA lipid particle) will be administered in accordance with the disclosed invention. , (ii) is receiving a TNA lipid particle (or a pharmaceutical composition comprising a TNA lipid particle) according to the described invention; or (i¡) has received a TNA lipid particle (or a pharmaceutical composition comprising a TNA lipid particle) in accordance with the described invention, unless the context and use of the phrase indicate otherwise. As used herein, the term suppress, decrease, interfere, inhibit and / or reduce (and similar terms) generally refers to the action of reducing, either directly or indirectly, a concentration, level, function, activity or behavior. in relation to the natural, expected or average condition, or in relation to a control condition. As used herein, the terms "synthetic AAV vector" and "synthetic AAV vector production" refer to an AAV vector and methods of synthetic production thereof in a completely cell-free environment. As used herein, the term systemic delivery is intended to refer to delivery of lipid particles that leads to broad biodistribution of an active agent such as an interfering RNA (e.g., siRNA) within an organism. Some management techniques ΜΛ / t / zuzz / uoy 101 may lead to systemic delivery of certain agents, but not others. Systemic delivery means that a useful, preferably therapeutic, amount of an agent is exposed to most parts of the body. To obtain broad biodistribution, a blood life expectancy is generally required such that the agent is not rapidly degraded or eliminated (e.g., by first-pass organs (liver, lung, etc.) or by rapid, nonspecific cell binding). before reaching a disease site distal to the site of administration. Systemic delivery of lipid particles (e.g., lipid nanoparticles) can be accomplished by any means known in the art, including, for example, intravenous, subcutaneous, and intraperitoneal. In a preferred embodiment, the systemic delivery of lipid particles (e.g., lipid nanoparticles) is by intravenous delivery. As used herein, the terms terminal resolution site and TRS are used interchangeably herein and are intended to refer to a region in which Rep forms a tyrosine-phosphodiester bond with the 5' thymidine that generates a 3' OH that serves as a substrate for DNA extension by a cellular DNA polymerase, for example, DNA pol delta or DNA pol epsilon. Alternatively, the Rep-thymidine complex may participate in a coordinated ligation reaction. As used herein, the terms therapeutic amount, therapeutically effective amount, an effective amount or pharmaceutically effective amount of an active agent (for example, a lipid particle with TNA as described herein) are used interchangeably to refer to a amount that is sufficient to provide the desired treatment benefit or effect, for example, inhibition of expression of a target sequence compared to the level of expression detected in the absence of a therapeutic nucleic acid. Suitable assays for determining the expression of a target gene or target sequence include, for example, examination of protein or RNA levels by techniques known to those skilled in the art, such as immunodot blotting, Northern technique, in situ hybridization, ELISA, immunoprecipitation, enzyme function, as well as phenotypic assays known to those skilled in the art. Dosage levels are based on a variety of factors, including type of injury, age, weight, sex, clinical status of the patient, severity of the condition, route of administration, and the particular active agent used. . Therefore, the dosage regimen can vary greatly, but can be determined routinely by a doctor using standard methods. Furthermore, the terms therapeutic amount, therapeutically effective amounts and pharmaceutically effective amounts include prophylactic or preventive amounts of the compositions of the disclosed invention. In prophylactic or preventive applications of the described invention, the pharmaceutical compositions or medicaments are administered to a patient susceptible to or otherwise at risk for a disease, disorder or condition in an amount sufficient to eliminate or reduce the risk, reduce the severity or slowing the onset of the disease, disorder or condition, including the biochemical, histological and / or behavioral symptoms of the disease, disorder or condition, its complications and intermediate pathological phenotypes that occur during the development of the disease, disorder or condition. It is generally preferred that a maximum dose be used, that is, the highest safe dose according to some medical criteria. The terms dose and dosage are used interchangeably herein. In one aspect of any of the aspects or embodiments herein, therapeutic amount, therapeutically effective amounts and pharmaceutically effective amounts refer to non-prophylactic or non-preventive applications. As used herein, the term therapeutic effect refers to a consequence of the treatment, the results of which are considered desirable and beneficial. A therapeutic effect may include, directly or indirectly, the arrest, reduction or elimination of a manifestation of the disease. A therapeutic effect may further include, directly or indirectly, arresting, reducing or eliminating the progression of a disease manifestation. For any therapeutic agent described herein, the therapeutically effective amount may initially be determined from preliminary in vitro studies and / or animal models. A therapeutically effective dose can further be determined from human data. The applied dose can be adjusted based on the relative bioavailability and potency of the administered compound. Adjusting the dosage to achieve maximum efficacy based on the methods described above and other known methods is within the capabilities of one skilled in the art. General principles for determining therapeutic efficacy, which can be found in Chapter 1 of The Pharmacological Basis of Therapeutics, 10th Edition, McGraw-Hill (New York) (2001) by Goodman and Gilman, incorporated herein by reference, are summarized as follows: continuation. Pharmacokinetic principles provide a basis for modifying a dosing regimen to obtain a desirable degree of therapeutic efficacy with a minimum of unacceptable adverse effects. In situations where the plasma concentration of the drug can be determined and related to the therapeutic window, additional guidance can be obtained to modify the dosage. As used herein, the terms “treat,” “treating,” and / or “treatment” include nullifying, inhibiting, slowing, or reversing the progression of a condition, improving the clinical symptoms of a condition, or preventing the occurrence of clinical symptoms. of a condition, thereby obtaining beneficial or desirable clinical results. Treatment further refers to achieving one or more of the following: (a) reducing the severity of the disorder; (b) limit the development of symptoms characteristic of the disorder or disorders being treated; (c) limit worsening of the symptoms characteristic of the disorder or disorders being treated; (d) limit recurrence of the disorder(s) in patients who have previously had the disorder(s); and (e) limit symptom recurrence in patients who were previously asymptomatic in relation to the disorder(s). In one aspect of any of the aspects or modalities herein, the terms "treat", "treating" and / or "treatment" include nullifying, inhibiting, slowing or reversing the progression of a condition, or improving the clinical symptoms of a condition. Beneficial or desirable clinical outcomes, such as pharmacological and / or physiological effects, include, but are not limited to, preventing the occurrence of the disease, disorder or condition in a subject who may be predisposed to the disease, disorder or condition but who still does not have it or exhibit symptoms of the disease (prophylactic treatment), relieve the symptoms of the disease, disorder or condition, reduce the extent of the disease, disorder or condition, stabilize (that is, not worsen) the disease, disorder or condition condition, prevent the spread of the disease, disorder or condition, delay or slow the progression of the disease, disorder or condition, improve or alleviate the disease, disorder or condition, and combinations of these, as well as prolong survival compared to expected survival if no treatment is received. As used herein, the terms “vector” or “expression vector” are intended to refer to a replicon, such as a plasmid, bacmid, phage, virus, virion or cosmid, to which another segment of DNA can be attached, i.e. an “insert”, “transgene” or “expression cassette” to achieve expression or replication of the joined segment (“expression cassette”) in a cell. A vector may be a nucleic acid construct designed for delivery to a host cell or for transfer between different host cells. As used herein, a vector may be of viral or non-viral origin in the final form. However, for the purposes of the present description, a vector generally refers to a synthetic AAV vector or a nicked ceDNA vector. Therefore, the term “vector” encompasses any genetic element that is capable of replicating when associated with the appropriate control elements and that can transfer gene sequences to cells. In some embodiments of any of the aspects and embodiments herein, a vector may be a recombinant vector or an expression vector. Groupings of elements or alternative embodiments of the invention described herein should not be construed as limitations. Each member of the group may be claimed or referenced individually or in any combination with other members of the group or other elements found herein. One or more members of a group may be included in, or removed from, a group for reasons of convenience and / or patentability. When such inclusion or deletion occurs, the description is hereby deemed to contain the modified group, so as to comply with the written description of all Markush groups used in the appended claims. In some embodiments of any of the aspects, the description described herein does not refer to a process for cloning human beings, processes for modifying the genetic identity of the MA / IZ / ¿U¿¿ / UO3 / 01 germ line of human beings, uses of human embryos for industrial or commercial purposes or processes to modify the genetic identity of animals that may cause them suffering without any substantial medical benefit to man or the animal, and also animals resulting from said processes. Other terms are defined herein within the description of the various aspects of the invention. All patents and other publications, including bibliographic references, issued patents, published patent applications and co-pending patent applications, cited throughout this application, are expressly incorporated herein by reference for the purpose of describing and disclosing, by For example, the methodologies described in such publications that could be used in connection with the technology described herein. These publications are provided solely for your description prior to the date of submission of this application. Nothing in this regard should be construed as an admission that inventors do not have the right to precede such a description by virtue of a prior invention or for any other reason. Any statements regarding the date or representation of the content of these documents are based on information available to applicants and do not constitute an admission as to the accuracy of the dates or content of these documents. The description of the embodiments of the description is not intended to be exhaustive or to limit the description to the precise form described. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the art will recognize. For example, while the method steps or functions are presented in a given order, alternative embodiments may perform the functions in a different order, or the functions may be performed substantially simultaneously. The teachings of the description provided herein may be applied to other procedures or methods as appropriate. The various embodiments described herein may be combined to provide additional embodiments. Aspects of the description may be modified, if necessary, to employ the compositions, functions and concepts of the above application and references to provide still other embodiments of the description. Furthermore, due to considerations of biological functional equivalence, some changes can be made to the protein structure without affecting the biological or chemical action in kind or quantity. These and other changes may be made to the description in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims. Specific elements of any of the above modalities may be combined or replaced by elements in other modalities. Furthermore, although the advantages associated with certain embodiments of the description were described in the context of these embodiments, other MA / t / zuzz / uoy 101 modalities may further exhibit such advantages, and not all modalities necessarily need to exhibit such advantages to be within the scope of the description. The technology described herein is further illustrated by the following examples which should in no way be construed as a further limitation. It should be understood that this invention is in no way limited to the particular methodology, protocols and reagents, etc., described herein and that these may vary. The terminology used herein is only intended to describe particular embodiments, and is not intended to limit the scope of the present invention, defined solely by the claims. II. LIPIDS In a first chemical embodiment, ionizable lipids of Formula (I) are provided: MA / t / zuzz / uoy / 01 or a pharmaceutically acceptable salt thereof, wherein: a is an integer ranging from 1 to 20 (for example, a is 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 , 18, 19 or 20); b is an integer ranging from 2 to 10 (for example, b is 2, 3, 4, 5, 6, 7, 8, 9, or 10); R1 is absent or selected from (C2-C2o)alkenyl, -C(0)O(C2-C2o)alkyl and cyclopropyl substituted with (C2-C2o)alkyl; and R2is alkyl(C2-C2o); In a second chemical embodiment, the ionizable lipid of Formula (I) is of Formula (II): or a pharmaceutically acceptable salt thereof, where c and d are each independently integers ranging from 1 to 8 (e.g., 1.2, 3, 4, 5, 6, 7 or 8), and where the remaining variables They are as described for Formula (I). In a third chemical embodiment, c and d in the ionizable lipid of Formula (I) or (II) or a pharmaceutically acceptable salt thereof, are each independently integers ranging from 2 to 8, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 4 to 8, 4 to 7, 4 to 6, 5 to 8, 5 to 7 or 6 to 8, where the remaining variables are as described for Formula (I) or (II ). In a fourth chemical embodiment, c in the ionizable lipid of Formula (I) or (II) is 2, 3, 4, 5, 6, 7 or 8, where the remaining variables are as described for Formula (I ) or the second or third chemical modalities. Alternatively, as part of a fourth chemical embodiment, c and d in the ionizable lipid of Formula (I) or (II) or a pharmaceutically acceptable salt thereof, are each independently 1, 3, 5 or 7, where the remaining variables are as described for Formula (I) or the second or third chemical embodiments. In a fifth chemical embodiment, d in the ionizable lipid of Formula (I) or (II) is 2, 3, 4, 5, 6, 7 or 8, where the remaining variables are as described for Formula (I ) or the second or third or fourth chemical modalities. Alternatively, as part of a fifth chemical embodiment, at least one of c and d in the ionizable lipid of Formula (I) or (II) or a pharmaceutically acceptable salt thereof is 7, wherein the remaining variables are as described for the Formula (I) or the second or third or fourth chemical modalities. In a sixth chemical embodiment, the ionizable lipid of Formula (I) is of Formula (III): EITHER Μλ. t / zuzz / uoy 101 or a pharmaceutically acceptable salt thereof, wherein the remaining variables are as described for Formula (I). In a seventh chemical embodiment, b in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is an integer ranging from 3 to 9, where the remaining variables are as described for Formula (I), or the second, third, fourth or fifth chemical embodiments. Alternatively, as part of a seventh chemical embodiment, b in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is an integer ranging from 3 to 8, from 3 to 7, from 3 to 6, 3 to 5, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 5 to 9, 5 to 8, 5 to 7, 6 to 9, 6 to 8, or 7 to 9, wherein the remaining variables are as described for Formula (I), or the second, third, fourth or fifth chemical embodiments. In another alternative, as part of a seventh chemical embodiment, b in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is 3, 4, 5, 6, 7, 8 , or 9, wherein the remaining variables are as described for Formula (I), or the second, third, fourth or fifth chemical embodiments. In an eighth chemical embodiment, the ionized lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof is an integer ranging from 2 to 18, where the remaining variables are as follows: described for Formula (I), or the second, third, fourth, fifth or seventh chemical modalities. Alternatively, as part of an eighth embodiment, in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is an integer ranging from 2 to 18, from 2 to 17 , from 2 to 16, 2 to 15, 2 to 14, 2 to 13, 2 to 12, 2 to 11,2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 3 to 18, 3 to 17, 3 to 16, 3 to 15, 3 to 14, 3 to 13, 3 to 12, 3 to 11, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 4 to 18, 4 to 17, 4 to 16, 4 to 15, 4 to 14, 4 to 13, 4 to 12, 4 to 11,4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 5 to 18, 5 to 17, 5 to 16, 5 to 15, 5 to 14, 5 to 13, 5 to 12, 5 to 11,5 to 10, 5 to 9, 25 to 8 , 5 to 7, 6 to 18, 6 to 17, 6 to 16, 6 to 15, 6 to 14, 6 to 13, 6 to 12, 6 to 11,6 to 10, 6 to 9, 6 to 8, 7 to 18, 7 to 17, 7 to 16, 7 to 15, 7 to 14, 7 to 13, 7 to 12, 7 to 11,7 to 10, 7 to 9, 8 to 18, 8 to 17, 8 to 16 , 8 to 15, 8 to 14, 8 to 13, 8 to 12, 8 to 11,8 to 10, 9 to 18, 9 to 17, 9 to 16, 9 to 15, 9 to 14, 9 to 13, 9 to 12, 9 to 11, 10 to 18, 10 to 17, 10 to 16, 10 to 15, 10 to 14, 10 to 13, 11 to 18, 11 to 17, 11 to 16, 11 to 15, 11 to 14 , 11 to 13, 12 to 18, 12 to 17, 12 to 16, 12 to 15, 12 to 14, 13 to 18, 13 to 17, 13 to 16, 13 to 15, 14 to 18, 14 to 17, 14 to 16, 15 to 18, 15 to 17, or 16 to 18, where the remaining varieties are as described for Formula (I), or the second, third, fourth, fifth or seventh chemical embodiments. In another alternative, as part of an eighth embodiment, a in the ionizable lipid of Formula (I), (II) or (III) is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18, where the remaining varieties are as described for Formula (I), or the second, third, fourth, fifth or seventh chemical embodiments. In a ninth chemical embodiment, R1 in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is absent or selected from (C5-Ci5)alkenyl, C(O)Oalkyl( C4-Ci8) and cyclopropyl substituted with alkyl (C4-Ci6), wherein the remaining varieties are as described for Formula (I), or the second, third, fourth, fifth, seventh, or eighth chemical embodiments. Alternatively, as part of a ninth chemical embodiment, R1 in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is absent or selected from (C5-Ci5)alkenyl, -C (O)O(C4-Ci6)alkyl and cyclopropyl substituted with (C4-C16)alkyl, wherein the remaining variants are as described for Formula (I), or the second, third, fourth, fifth, seventh or eighth embodiments chemicals. Alternatively, as part of a ninth chemical embodiment, R1 in the ionizable lipid of formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is absent or selected from (C5-C12)alkenyl, ma. t / zuzz / uoy 101 C(O)Oalkyl(C4-Ci2) and cyclopropyl substituted with alkyl(C4-Ci2), wherein the remaining variables are as described for Formula (I), or the second, third, fourth, fifth, seventh or eighth chemical modalities. In another alternative, as part of a ninth chemical embodiment, R1 in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is absent or selected from alkenyl (C5-Cio), -C(0)Oalkyl(C4-Cio) and cyclopropyl substituted with alkyl(C4-Cio), where the remaining variables are as described for Formula (I), or the second, third, fourth, fifth, seventh or eighth chemical modalities. In a tenth chemical embodiment, R1 is alkenyl Cw, where the remaining variables are as described in any of the previous embodiments. In an eleventh chemical embodiment, the alkyl in C(0)Oalkyl(C2-C2o), -C(O)O alkyl(C4Ci8)alkyl, -C(O)O alkyl(C4-Ci2)), or - 0(0)0(C4-Cio)alkyl of R1 in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is an unbranched alkyl, wherein the remaining variables are as described in any of the previous modalities. In a chemical embodiment, R1 is -C(O)O(C9 alkyl). Alternatively, in an eleventh chemical embodiment, the alkyl in -C(O)Oalkyl(C4-Ci8), -0(0)0 alkyl(C4-Ci2), or -0(0)0 alkyl(C4-Cio) of R1 in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is a branched alkyl, wherein the remaining variables are as described in any of the above chemical embodiments. In a chemical embodiment, R1 is -C(O)O(C17 alkyl), where the remaining variables are as described in any of the previous chemical embodiments. In a twelfth chemical embodiment, R1 in the ionizable lipid of Formula (I), (II) or (III) or a pharmaceutically acceptable salt thereof, is selected from any group listed in Table 1 below, wherein the wavy bond in each of the groups indicates the point of attachment of the group to the rest of the lipid molecule, and where the remaining variables are as described for Formula (I), or the second, third, fourth, fifth, seventh or eighth chemical modalities. The present description also contemplates the combination of any of the groups R1 in Table 1 with any of the groups R2 in Table 2, where the remaining variables are as described for Formula (I), or the second, third, fourth, fifth, seventh or eighth chemical modalities. In a thirteenth chemical embodiment, R2 in the ionizable lipid of Formula (I) or a pharmaceutically acceptable salt thereof, is selected from any group listed in Table 2 below, where the wavy bond in each of the groups indicates the point binding of the group to the rest of the lipid molecule, and wherein the remaining variables are as described for Formula (I), or the seventh, eighth, ninth, tenth or eleventh chemical embodiments. MA / lz / ¿u¿¿ / uoy 101 Specific examples are provided in Table 3, the exemplification section below and are included as part of a fourteenth chemical embodiment of ionizable lipids of Formula (I) herein. Also included are pharmaceutically acceptable salts, as well as ionized and neutral forms. Table 3. Illustrative ionizable lipids of the description. Yes 1-(Heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxy)phenyl))) nonanedioate lipid 1 acetoxy¡)ethyl)p¡peridin-1 -yl)ethyl)disulfanoyl)ethyl)piperidín-4-yl)ethoxy)-2-oxoethyl)phenyl) Yo Lipid 2 nonanedioate 1-(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-((5- (nonylox¡)-5oxopentanoyl)ox¡)phen¡l)acetoxy)ethyl)p¡per¡din-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phen it) EITHER .· Ί 'o Lipid 3 nonanedioate 1-(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-((9- (nonylox¡)-9oxononanoyl)oxy)phen¡l)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) either Jo either J. Ό Lipid 4 nonanedioate 1-(heptadecan-9-yl) 9-(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-((5- (nonylox¡)-5oxopentanoyl)ox¡)phen¡l)acetoxy)ethyl) piperidin-1 -i l)ethyl)disulfanoyl)et¡ l)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) 0'1,01-((((((disulfanedi¡lbis(ethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1) lipid 5-di(nonanedioate) diyl))bis(oxy)) bis(2-oxoethane-2,1-di¡l))bs(4,1-phenylene)) 9,9'-di(heptadecan-9-yl) 1 -(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-(oleoyloxy)phenyl)acetoxy)ethyl)piperidin-1-yl) lipid 6-nonanedioate )ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl)9-(undecan-3-yl) 1 -(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-(oleoyloxy)phenyl)acetoxy¡)ethyl)piperidin)-1 lipid 7-nonanedioate ¡l)ethyl)disulfanoyl)ethyl)p¡peridin-4-yl)ethoxy)-2-oxoethyl)pheníl)9-(tridecane-5-ilo) 1 -(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-(oleoyloxy)phenyl)acetoxy¡)ethyl)piperidin-1yl) nonanedioate ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl)9-(pentadecan-7-yl) Lipid 9-nonyl 9-nonanedioate 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-((9-oxo- 9-(undecan)-3yloxy)nonanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)ethyl)phenyl) Μλ. t / zuzz / uoy 101 Lipid 10 1-nonyl nonanedioate 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-((9-oxo- 9-(tridecane)-5yloxy)nonano¡l)oxy)phen¡l)acetoxy)ethyl)p¡peridin-1 -yl)ethyl)disulfanoyl)ethyl)piper¡din-4-yl)ethoxy)ethyl)phenyl) Lipid 11 1-nonyl nonanedioate 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-((9-oxo- 9-(pentadecane)-7¡lox¡)nonanoyl)oxy)phen¡l)acetoxy¡)ethyl)piper¡din-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)ethyl) phenyl) Lipid 12 nonanedioate 1 -(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-(((9Z ,12Z)-octadeca-9,12dienoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)dísulfanoyl)ethyl)piperidín-4-yl)ethoxy)-2-oxoethyl )phenyl) ivia / t / zuzz / uoy roí Lipid 13 nonanedioate 1 -(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-((8- (2octylcyclopropyl)octanoyl)oxy)phenyl)acetoxy)ethyl)piperídin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2- Lipid 14 nonanedioate 1-(heptadecan-9-yl) 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4( stearoyloxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)ethyl)phenyl) Lipid 15 nonanedioate 1-(heptadecan-9-yl) 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4( undecanoyloxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)ethyl)phenyl) .0. . T or .0. . You or Lipid 16 1-(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(nonanoyloxy)phenyl))) nonanedioate acetoxy¡)et¡l)piper¡d¡n-1 -¡l)ethyl¡l)d¡sulfanoyl)ethyl)piper¡d¡n-4-yl)ethoxy)-2-oxoethyl)phen it) or Lipid 17 1-nonyl nonanedioate 9-(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-((9-((3-octylundecyl)) oxy)-9)oxononanoyl)oxy)phenyl)acetoxy¡)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) OR G J... ...ll... ... ... ... . 'W- 1 -(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-((7-(heptadecan-9-yloxy))-7oxoheptanoyl)) nonanedioate 18 nonanedioate )oxy)phen¡l)acetoxy¡)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl)9nonyl Lipid 19 1-nonyl nonanedioate 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-((9-((3-octllundecyl)) ox¡)-9)oxononanoyl)oxy)phenyl)acetoxy)ethyl)p¡perídin-1 -¡l)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy¡)-2-oxoethyl)phen it) Lipid 20 1-nonyl nonanedioate 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-((7-((3-octylundecyl)) ox¡)-7)oxoheptanoyl)oxy)phenyl)acetoxy)ethyl)piper¡din-1 -yl)ethyl)d¡sulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) In another aspect, lipids of Formula (la), (Ib), or (le) are contemplated herein: or a pharmaceutically acceptable salt thereof, wherein Rq and Rz are each independently an aliphatic group (including alkyls, alkenyls, alkynyls, cycloalkyls, heterocyclyls) or an aryl group, where the remaining variables are as described above in any of the embodiments previous chemistries. In one embodiment, Rq and Rz are each independently hydrogen or Οι-Οθ alkyl, where the remaining variables are as described above in any of the above chemical embodiments. The LNPs, compositions, methods of use, etc., further apply to lipids of Formula (la), (Ib), or (le). The lipids of Formula (la), (Ib) or (le) can be prepared, for example, the lipid of Formula (I), by treatment with chloromethane (CH3CI) in acetonitrile (CH3CN) and chloroform (CHCI3). Furthermore, a lipid of Formula (II) or (III), or any of the illustrative lipids described herein, can be converted to the corresponding quaternary lipids (all contemplated in this description), for example, the lipid of Formula (I) by treatment with chloromethane (CH3CI) in acetonitrile (CH3CN) and chloroform (CHCI3). Lipid particles (LNPs), or pharmaceutical compositions thereof, comprising an ionizable lipid described herein and a capsid-free non-viral vector (e.g., ceDNA) can be used to deliver the capsid-free non-viral DNA vector. to a target site of interest (e.g., cell, tissue, organ and the like). In one embodiment of any of the aspects or embodiments herein, a formulation of lipid particles (e.g., lipid nanoparticles) is prepared and loaded with the TNA. In one embodiment, a formulation of lipid particles (e.g., lipid nanoparticles) is prepared and loaded with ceDNA obtained through the process described in international application PCT / US2018 / 050042, filed September 7, 2018, which is incorporated for reference in its entirety herein. This can be achieved by high-energy mixing of ethanolic lipids with aqueous TNA such as ceDNA at low pH, which protonates the lipid and provides a favorable energy for ceDNA / lipid association and particle nucleation. The particles can be further stabilized by aqueous dilution and removal of the organic solvent. The particles can be concentrated to the desired level. Typically, lipid particles (e.g., lipid nanoparticles) are prepared in a ratio of total lipid to nucleic acid (mass or weight) of about 10:1 to 60:1. In some embodiments of any of the aspects or embodiments herein, the ratio of lipids to nucleic acid (mass / mass ratio; w / w ratio) may be in the range of about 1:1 to about 60:1, from about 1:1 to about 55:1, about 1:1 to about 50:1, about 1:1 to about 45:1, about 1:1 to about 40:1, about 1:1 to about 35 :1, about 1:1 to about 30:1, about 1:1 to about 25:1, about 10:1 to about 14:1, about 3:1 to about 15:1, about 4 :1 to about 10:1, about 5:1 to about 9:1, about 6:1 to about 9:1; from about 30:1 to about 60:1. According to some embodiments of any of the aspects or embodiments herein, lipid particles (e.g., lipid nanoparticles) are prepared in a ratio of nucleic acid (mass or weight) to total lipid of about 60:1. According to some embodiments of any of the aspects or embodiments herein, lipid particles (e.g., lipid nanoparticles) are prepared in a ratio of nucleic acid (mass or weight) to total lipid of about 30:1. The amounts of lipid and nucleic acid can be adjusted to provide a desired N / P ratio, for example, an N / P ratio of 3, 4, 5, 6, 7, 8, 9, 10,11,12,13,14 15,16, 17,18,19, 20 or higher. Generally, the total lipid content of the lipid particle formulation can range from about 5 mg / mL to about 30 mg / rriL. In some embodiments of any of the aspects or embodiments herein, the lipid nanoparticle comprises an agent for condensing and / or encapsulating the nucleic acid cargo, such as ceDNA. Such an agent is further referred to herein as a condensing or encapsulating agent. Without limitations, any compound known in the art can be used to condense and / or encapsulate nucleic acids as long as it is not fusogenic. In other words, an agent capable of condensing and / or encapsulating nucleic acid cargo, such as ceDNA, but having little or no fusogenic activity. Without wishing to be limited by theory, a condensing agent may have some fusogenic activity when it does not condense / encapsulate a nucleic acid, such as ceDNA, but a lipid nanoparticle encapsulating a nucleic acid, formed with said condensing agent may not be fusogenic. Generally, an ionizable lipid or a cationic lipid is typically employed to condense the nucleic acid cargo, for example, ceDNA at low pH and to drive fusogenicity and membrane association. Generally, cationic lipids are lipids that comprise at least one amino group that is positively charged or protonated under acidic conditions, for example, at a pH of 6.5 or less. The cationic lipids may also be ionizable lipids, for example, ionizable cationic lipids. By a non-fusogenic ionizable lipid is meant an ionizable lipid that can condense and / or encapsulate nucleic acid payload, such as ceDNA, but has no, or very little, fusogenic activity. In one embodiment of any of the aspects or embodiments herein, the ionizable lipid may comprise 20-90% (mol) of the total lipid present in the lipid particles (e.g., lipid nanoparticles). For example, the molar content of ionizable lipids may be 2070% (mol), 30-60% (mol), 40-60% (mol), 40-55% (mol), or 45-55% (mol). of the total lipids present in the lipid particle (for example, lipid nanoparticles). In some embodiments of any of the aspects or embodiments herein, the ionizable lipid comprises MA / lz / ¿u¿¿ / uoy / ΟΊ about 50 mol % to about 90 mol % of the total lipid present in the lipid particles (e.g., lipid nanoparticles). In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) may further comprise a non-cationic lipid. The non-cationic lipid may serve to increase fusogenicity and also increase the stability of the LNP during formation. Noncationic lipids include antipathic lipids, neutral lipids, and anionic lipids. Accordingly, the non-cationic lipid may be an uncharged neutral, zwitterionic or anionic lipid. Generally, non-cationic lipids are used to improve fusogenicity. Illustrative non-cationic lipids include, but are not limited to, distearoyl-sn-glycerophosphoethanolamine, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC) dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoylphosphatidyl lina (POPC ), palmitoylphosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoyl-phosphoethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE ), monomethyl-phosphatidylethanolamine (such as 16-O-monomethyl PE), dimethylphosphatidylethanolamine (such as 16O-dimethyl PE), 18-1-trans PE, 1-stearol-2-oleoylphosphatidiethanolamine (SOPE), hydrogenated soy phosphatidylcholine (HSPC) (POPG), dielaidoyl-phosphatidylethanolamine (DEPE), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE); 1,2-difantoyl-sn-glycero-3phosphoethanolamine (DPHyPE); lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidicazide, cerebrosides, dicetylphosphate, lysophosphatidylcholine, dilinoleophosphatidylcholine, or mixtures thereof. It should be understood that other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids may also be used. The acyl groups in these lipids are preferably acyl groups derived from fatty acids having C10-C24 carbon chains, for example, lauroyl, myristoyl, palmitoyl, stearoyl or oleoyl. Other examples of non-cationic lipids suitable for use in lipid particles (e.g., lipid nanoparticles) include non-phosphorous lipids, e.g., stearylamine, dodecylamine, hexadecylamine, acetyl palmitate, glycerolricinoleate, hexadecyl stearate, isopropyl myristate, polymers amphoteric acrylic, triethanolamine lauryl sulfate, polyethyloxylated fatty acid amides of alkyl aryl sulfate, dioctadecyldimethylammonium bromide, ceramide, sphingomyelin and the like. MA / IZ / ¿U¿¿ / UO3 / ΟΊ In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid is a phospholipid. In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid is selected from the group consisting of DSPC, DPPC, DMPC, DOPC, POPO, DOPE and SM. In some embodiments of any of the aspects or embodiments herein, the non-cationic lipid is DSPC. In other embodiments, the non-cationic lipid is DOPC. In other embodiments, the non-cationic lipid is DOPE. In some embodiments of any of the aspects or embodiments herein, the non-cationic lipid may comprise from 0 to about 20% (mol) of the total lipid present in the lipid nanoparticle. In some embodiments of any of the aspects or embodiments herein, the non-cationic lipid content is 0.5-15% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the non-cationic lipid content is 5-12% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the non-cationic lipid content is 5-10% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid content is approximately 6% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid content is approximately 7.0% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid content is approximately 7.5% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid content is approximately 8.0% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid content is approximately 9.0% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the non-cationic lipid content is approximately 10% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the non-cationic lipid content is approximately 11% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). Illustrative non-cationic lipids are described in PCT Publication WO2017 / 099823 and United States Patent Publication US2018 / 0028664, the contents of which are incorporated herein by reference in their entirety. In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) may further comprise a component, such as an ester, to provide membrane integrity and stability of the lipid particle. In one embodiment of any of the aspects or embodiments herein, an illustrative ester that may be used in the lipid particle is cholesterol or a derivative thereof. Non-limiting examples of cholesterol derivatives include polar analogs, such as 5a-cholestanol, 5βcoprostanol, cholesteryl-(2'-hydroxy)-ethyl ether, cholesteryl-(4'-hydroxy)-but. ether and 6-ketocholestanol; non-polar analogs, such as 5a-cholestane, cholestenone, 5a-cholestanone, 5β-οοΙβ3ΐ3ηοη3 and cholesteryl decanoate; and mixtures of these. In some embodiments of any of the aspects or embodiments herein, the cholesterol derivative is a polar analog such as cholesteryl-(4-hydroxyl)-butyl ether. In some embodiments of any of the aspects or embodiments herein, the cholesterol derivative is cholestril hemisuccinate (CHEMS). Illustrative cholesterol derivatives are described in PCT Publication W02009 / 127060 and United States Patent Publication US2010 / 0130588, the contents of which are incorporated herein by reference in their entirety. In one embodiment of any of the aspects or embodiments herein, the component that provides membrane integrity, such as an ester, may comprise 0-50% (mol) of the total lipid present in the lipid particle (e.g., nanoparticle lipid). In some embodiments of any of the aspects or embodiments herein, such component is 2050% (mol) of the total lipid content of the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, such component is 30-40% (mol) of the total lipid content of the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, such component is 35-45% (mol) of the total lipid content of the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, such component is 38-42% (mol) of the total lipid content of the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the lipid particle (e.g., lipid nanoparticle) may further comprise a polyethylene glycol (PEG) or a conjugated lipid molecule. Generally, these are used to inhibit the aggregation of a lipid particle (e.g., lipid nanoparticle) and / or provide spherical stabilization. Illustrative conjugated lipids include, but are not limited to, PEG-lipid conjugates, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (such as ATTAlipid conjugates), cationic polymer-lipid (CPL) conjugates, and mixtures. of these. In some embodiments of any of the aspects or embodiments herein, the conjugated lipid molecule is a PEG-lipid conjugate, for example, a lipid-(methoxypolyethylene glycol) conjugate. In some other embodiments, the conjugated lipid molecule is a PEG-lipid conjugate, for example, a PEG2000DMG (dimyristoylglycerol). Illustrative PEG-lipid conjugates include, but are not limited to, PEG-diacylglycerol (DAG) (such as l-(monomethoxy-polyethylene glycol)-2,3-dimyristo¡lglycerol (PEG-DMG)), PEG-dialkyloxypropyl (DAA), PEG-phospholipid, PEG-ceramide (Cer), a pegylated phosphatidylethanolamine (PEG-PE), PEG diacylglycerol succinate (PEGS-DAG) (such as 4-0-(2',3'di(-butanedioate) tetradecane¡lox¡)prop¡l-1-0-(w-methoxy¡(polyethoxy¡)ethyl) (PEG-S-DMG)), PEG dialkoxypropylcarbam, N-(carbonyl-methoxy¡pol sodium salt ethylene glycol 2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine, or a mixture thereof. Additional illustrative PEG-lipid conjugates are described, for example, in US5,885,613, US6,287,591 , US2003 / 0077829, US2003 / 0077829, US2005 / 0175682, US2008 / 0020058, US2011 / 0117125, US2010 / 0130588, US2016 / 0376224 and US2017 / 0119904, the contents of which are incorporated herein by reference in their entirety. In one embodiment of any of the aspects or embodiments herein, the PEG-DAA conjugate may be, for example, PEG-dilauryloxypropyl, PEG-dimyristyloxypropyl, PEGdipalmityloxypropyl or PEG-distearyloxypropyl. The PEG-lipid may be one or more of PEG-DMG, PEG-dilaurylglycerol, PEG-dipalmitoylglycerol, PEG-dysterylglycerol, PEG-dilaurylglycamide, PEGdimyristylglycamide, PEG-dipalmitoylglycerol, PEG-dysterylglycerol, PEG-cholesterol (l-[8'- (Colest-5en-3[beta]-ox¡)carboxamido-3',6'-dioxaoctan¡l]carbamo¡l-[omega]-met¡l-poly(ethylene glycol), PEG- DMB (3,4-d¡tetradecox¡benzyl-[omega]-methyl-poly¡(ethyleneglycol)ether), and l,2-dimyristoyl-sn-glycero-3phosphoethanolamine-N-[methoxy¡( polyethylene glycol)-2000]. In one embodiment of any of the aspects or embodiments herein, the PEG-lipid can be selected from the group consisting of PEGDMG, 1,2-d¡myr¡sto¡l -sn-glycero-3-phosphoethanolamine-N-[methoxy¡(polyethyleneglycol)-2000]. In one embodiment of any of the aspects or embodiments herein, lipids conjugated to a molecule other than PEG may also be used instead of PEG-lipid. For example, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (such as ATTA-lipid conjugates), and cationic polymer-lipid (CPL) conjugates can be used instead of or in addition to the PEG-lipid conjugate. . Illustrative conjugated lipids, i.e., PEG-lipids, (POZ)-lipid conjugates, ATTA-lipid conjugates and cationic polymers-lipids are described in PCT patent application publications WO1996 / 010392, WO1998 / 051278, W02002 / 087541, W02005 / 026372, W02008 / 147438, W02009 / 086558, WO2012 / 000104, WO2017 / 117528, WO2017 / 099823, WO2015 / 199952, WO2017 / 004143, WO2015 / 095346, WO2012 / 000104, WO2012 / 000104 and WO2010 / 006282, United States Patent Application Publications US2003 / 0077829, US2005 / 0175682, US2008 / 0020058, US2011 / 0117125, US2013 / 0303587, US2018 / 0028664, US2015 / 0376115, US2016 / 0376224, US2016 / 0317458, US2013 / 0303587, US2013 / 0303587 and US20110123453, and United States patents US5,885,613, US6,287,591, US6,320,017 and US6,586,559, the contents of which are incorporated herein by reference in their entirety. In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate is present in a molar ratio of about 0% to about 20% in the lipid nanoparticle. In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is 0.5 to 10% (mol) in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is 1-5% (mol) in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is 1-3% (mol) in the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the PEG-lipid conjugate content is approximately 1.5% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is about 2% (mol) in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is about 2.5% (mol) in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is approximately 3% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is about 3% (mol) in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the PEG-lipid conjugate content is about 3.5% (mol) in the lipid particle (e.g., lipid nanoparticle). In some embodiments of any of the aspects or embodiments herein, the conjugated lipid, such as the PEG-lipid conjugate or the PEG-gylated lipid, is present at a molar percentage greater than about 2.0% of the total lipid in the nanoparticle. lipid, for example, about 2.1%, or 2.2%, or 2.3%, or 2.4%, or about 2.5% to about 10%; or about 2.1%, or 2.2%, or 2.3%, or 2.4%, or about 2.5% to about 7.5%; about 2.1%, or 2.2%, or 2.3%, or 2.4%, or about 2.5% to about 5%; about 3% to about 5%; about 3% to about 4.5%; about 3% to about 4%; about 3.5% to about 5%; 3.5% to 4.5%, from 2.5% to 4%; about 2.5% to about 3.5%, or about 2.5% to about 3%. It is understood that the molar ratios of the described ionizable lipid to the non-cationic lipid, the ester and the PEG-conjugated lipid may vary as necessary. For example, the lipid particle (e.g., lipid nanoparticle) may comprise 30-70% ionizable lipid per mole or by total weight of the composition, 0-60% cholesterol per mole or by total weight of the composition, 030 % non-cationic lipid per mole or by total weight of the composition and 1 -10% of PEG-conjugated lipid per mole or by total weight of the composition. In one embodiment of any of the aspects or embodiments herein, the composition comprises 40-60% ionizable lipids per mole or by total weight of the composition, 30-50% cholesterol per mole or by total weight of the composition, 5-15% of non-cationic lipid per mole or by total weight of the composition and 1-5% of the PEG-conjugated lipid per mole or by total weight of the composition. In one embodiment of any of the aspects or embodiments herein, the composition has 40-60% ionizable lipids per mole or by total weight of the composition, 30-40% cholesterol per mole or by total weight of the composition, and 5-10% non-cationic lipid, per mole or by total weight of the composition and 15% PEG-conjugated lipid per mole or by total weight of the composition. The composition may contain 60-70% ionizable lipids per mole or by total weight of the composition, 25-35% cholesterol per mole or by total weight of the composition, 5-10% non-cationic lipids per mole or by total weight of the composition and 0-5% of PEG-conjugated lipid per mole or per total weight of the composition. The composition may further contain up to 45-55% ionizable lipids per mole or by total weight of the composition, 35-45% cholesterol per mole or by total weight of the composition, 2-15% non-cationic lipid per mole or per total weight of the composition, and 1-5% of PEG-conjugated lipid per mole or per total weight of the composition. The formulation may further be a lipid nanoparticle formulation, for example, comprising 8-30% ionizable lipids per mole or by total weight of the composition, 5-15% non-cationic lipids per mole or by total weight of the composition , and 0-40% cholesterol per mole or by total weight of the composition; 425% ionizable lipids per mole or by total weight of the composition, 4-25% non-cationic lipids per mole or by total weight of the composition, 2 to 25% cholesterol per mole or by total weight of the composition, 10 at 35% conjugated lipids per mole or by total weight of the composition, and 5% cholesterol per mole or by total weight of the composition; or 2-30% ionizable lipids per mole or by total weight of the composition, 2-30% non-cationic lipids per mole or by total weight of the composition, 1 to 15% cholesterol per mole or by total weight of the composition composition, 2 to 35% PEG-conjugated lipid per mole or by total weight of the composition, and 1-20% cholesterol per mole or by total weight of the composition; or even up to 90% ionizable lipids per mole or by total weight of the composition and 2-10% non-cationic lipids per mole or by total weight of the composition, or even 100% ionizable lipids per mole or by weight total composition. In some embodiments of any of the aspects or embodiments herein, the lipid particle formulation comprises an ionizable lipid, a non-cationic phospholipid, cholesterol and a lipid. MA / IZ / ¿U¿¿ / UO3 / ΟΊ PEGylated (conjugated lipid) in a molar ratio of approximately 50:10:38.5:1.5. In some embodiments of any of the aspects or embodiments herein, the lipid particle formulation comprises an ionizable lipid, a non-cationic phospholipid, cholesterol and a PEGylated lipid (conjugated lipid) in a molar ratio of approximately 50:10:38: 2. In some embodiments of any of the aspects or embodiments herein, the lipid particle formulation comprises an ionizable lipid, a non-cationic phospholipid, cholesterol and a PEGylated lipid (conjugated lipid) in a molar ratio of approximately 50:10:37: 3. In one embodiment of any of the aspects or embodiments herein, the lipid particle formulation (e.g., lipid nanoparticles) comprises ionizable lipid, non-cationic phospholipid, cholesterol and a PEGylated lipid (conjugated lipid) in a molar ratio of about 50 :7:40:3. In one embodiment of any of the aspects or embodiments herein, the lipid particle formulation (e.g., lipid nanoparticles) comprises ionizable lipid, non-cationic phospholipid, cholesterol and a PEGylated lipid (conjugated lipid) in a molar ratio of about 50 :8:40:2. In one embodiment of any of the aspects or embodiments herein, the lipid particle formulation (e.g., lipid nanoparticles) comprises ionizable lipid, non-cationic phospholipid, cholesterol and a PEGylated lipid (conjugated lipid) in a molar ratio of about 50 :9:39:2. In one embodiment of any of the aspects or embodiments herein, the lipid particle formulation (e.g., lipid nanoparticles) comprises ionizable lipid, non-cationic phospholipid, cholesterol and a PEGylated lipid (conjugated lipid) in a molar ratio of about 50 :9:38:3. In one embodiment of any of the aspects or embodiments herein, the lipid particle (e.g., lipid nanoparticle) comprises ionizable lipid, non-cationic lipid (e.g., phospholipid), a sterol (e.g., cholesterol) and a PEGylated lipid. (conjugated lipid), where the lipid molar ratio ranges from 20 to 70 mole percent for the ionizable lipid, with a target of 30-60, the noncationic lipid mole percent ranges from 0 to 30, with a target of 0 to 15, the molar percentage of sterol varies from 20 to 70, with a target of 30 to 50, and the molar percentage of PEGylated lipid (conjugated lipid) varies from 1 to 6, with a target of 2 to 5. Lipid nanoparticles (LNPs) comprising ceDNA are described in international application PCT / US2018 / 050042, filed September 7, 2018, which is incorporated herein in its entirety and contemplated for use in methods and compositions, such as is described herein. The particle size of lipid particles (e.g. lipid nanoparticles) can be determined by quasi-elastic light scattering using a Malvern Zetasizer Nano ZS (Malvern, UK), and has a diameter of approximately 50-150 nm, approximately 5595nm or approximately 70-90nm. MA / lz / ¿u¿¿ / uoy l OI The pKa of the formulated ionizable lipids can be correlated with the efficiency of LNPs for nucleic acid delivery (see Jayaraman et al., Angewandte Chemie, International Edition (2012), 51 (34), 8529-8533; Semple et al., Nature Biotechnology 28, 172-176 (20 1 0), both incorporated by reference in their entirety). In one embodiment of any of the aspects or embodiments herein, the pKa of each ionized lipid is determined in lipid nanoparticles by using a 2-(p-toluidino)-6naphthalenesulfonic acid (TNS) fluorescence-based assay. Lipid nanoparticles comprising ionizable lipid / DSPC / cholesterol / PEG-lipid (50 / 10 / 38.5 / 1.5 mol%) in PBS at a total lipid concentration of 0.4 mM can be prepared by using the online process as described in the present and elsewhere. TNS can be prepared as a 100 mM stock solution in distilled water. The vesicles can be diluted to 24 mM lipids in 2 mL of buffer solutions containing 10 mM HEPES, 10 mM MES, 10 mM ammonium acetate, 130 mM NaCl, where the pH varies from 2.5 to 11. A aliquot of the TNS solution to obtain a final concentration of 1 mM and, after mixing by vortexing, the fluorescence intensity is measured at room temperature on an SLM Aminco Series 2 luminescence spectrophotometer by using excitation wavelengths. and emission of 321 nm and 445 nm. A best-fit sigmoidal analysis can be applied to the fluorescence data, and the pKa is measured as the pH that generates the half-maximal fluorescence intensity. In one embodiment of any of the aspects or embodiments herein, relative activity can be determined by measuring luciferase expression in the liver 4 hours after administration by tail vein injection. The activity is compared at a dose of 0.3 and 1.0 mg ceDNA / kg and is expressed as ng luciferase / g liver measured 4 hours after administration. Without limitation, a lipid particle (e.g., lipid nanoparticle) of the disclosure includes a lipid formulation that can be used to deliver a capsid-free non-viral DNA vector to a target site of interest (e.g., cell, tissue, organ and similar). Generally, the lipid particle (e.g., lipid nanoparticle) comprises capsid-free non-viral DNA vector and an ionizable lipid or a salt thereof. In one embodiment of any of the aspects or embodiments herein, the lipid particle (e.g., lipid nanoparticle) comprises an ionizable lipid / non-cationic lipid / sterol / conjugated lipid in a molar ratio of 50:10:38.5:1.5. . In one embodiment of any of the aspects or embodiments herein, the disclosure provides a lipid particle formulation (e.g., lipid nanoparticle) comprising phospholipids, lecithin, phosphatidylcholine and phosphatidylethanolamine. iviA / t / zuzz / uoy roí III. THERAPEUTIC NUCLEIC ACID (TNA) The present disclosure provides a lipid-based platform for delivering therapeutic nucleic acid (TNA). Non-limiting examples of RNA-based therapeutic agents include mRNA, antisense RNA and oligonucleotides, ribozymes, aptamers, RNA interference (RNAi), Dicer substrate dsRNA, small hairpin RNA (shRNA), RNA interference asymmetric (iaRNA), microRNA (miRNA). Non-limiting examples of DNA therapeutics include minicircle DNA, minigene, viral DNA (e.g., AAV or lentivirus genome) or non-viral DNA vectors, closed-end linear duplex DNA (ceDNA / CELiD), plasmids, bacmids, doggybone™ DNA vectors, minimalist immunologically defined gene expression (MIDGE) vector, non-viral ministrand DNA vector (covalently closed linear DNA vector) or minimal dumbbell DNA vector (dumbbell DNA) . As such, aspects of the present disclosure generally provide ionizable lipid particles (e.g., lipid nanoparticles) comprising a TNA. Therapeutic nucleic acids Illustrative therapeutic nucleic acids of the present disclosure may include, but are not limited to, minigenes, plasmids, minicircles, small interfering RNA (siRNA), microRNA (miRNA), antisense oligonucleotides (ASO), ribozymes, closed-end double-stranded DNA ( for example, ceDNA, CELiD, covalently closed linear DNA (mini-strand), doggybone™, protelomer closed-end DNA or dumbbell linear DNA), Dicer substrate dsRNA viral vectors, small hairpin RNA (shRNA), asymmetric interfering RNA (iaRNA), microRNA (miRNA), mRNA, tRNA, rRNA and DNA, viral RNA vector and any combination of these. Further contemplated in the present invention as nucleic acid therapeutic agents is siRNA or miRNA that can downregulate intracellular levels of specific proteins through a process called RNA Interference (RNAi). Once siRNA or miRNA is introduced into the cytoplasm of a host cell, these double-stranded RNA constructs can bind to a protein called RISC. The sense strand of the siRNA or miRNA is removed by the RISC complex. The RISC complex, when combined with complementary mRNA, cleaves the mRNA and releases the cut strands. RNAi is by induction of specific destruction of the mRNA resulting in the inhibition of a corresponding protein. Antisense oligonucleotides (ASOs) and ribozymes that inhibit the translation of mRNA into protein may be nucleic acid therapeutic options. For antisense constructs, these single-stranded deoxynucleic acids have a sequence complementary to the mRNA sequence of the target protein and are capable of binding to the mRNA through Watson-Cñck base pairing. This binding prevents translation of a target mRNA and / or triggers RNaseH degradation of the mRNA transcript. As a result, the antisense oligonucleotide has a higher specificity of action (i.e., inhibition of a specific disease-related protein). In any of the methods and compositions provided herein, the therapeutic nucleic acid (TNA) may be a therapeutic RNA. Said therapeutic RNA may be an inhibitor of mRNA translation, RNA interference agent (RNAi), catalytically active RNA molecule (ribozyme), transfer RNA (tRNA) or an RNA that binds to an mRNA transcript (ASO). ), protein or other molecular ligand (aptamer). In any of the methods provided herein, the RNAi agent may be a double-stranded RNA, single-stranded RNA, microRNA, short interfering RNA, short hairpin RNA, or a triple helix-forming oligonucleotide. In any of the method compositions provided herein, the therapeutic nucleic acid (TNA) may be a therapeutic DNA such as double-stranded DNA with closed ends (e.g., ceDNA, CELiD, covalently closed linear DNA (mini-strand), doggybone™ , protelomeric DNA with closed ends, linear dumbbell DNA, plasmid, minicircle or similar). Some embodiments of the disclosure are based on methods and compositions comprising closed-end linear double-stranded (ceDNA) that can express a transgene (e.g., a therapeutic nucleic acid). The ceDNA vectors as described herein do not have packaging restrictions imposed by the limiting space within the viral capsid. ceDNA vectors represent a viable eukaryotic-produced alternative to prokaryote-produced plasmid DNA vectors. The ceDNA vectors preferably have a linear and continuous structure rather than a non-continuous structure. The linear, continuous structure is believed to be more stable against attack by cellular endonucleases, as well as less likely to recombine and cause mutagenesis. Therefore, a ceDNA vector in the linear and continuous structure is a preferred embodiment. The linear continuous intramolecular single-strand duplex ceDNA vector may have covalently linked terminal ends, without sequences encoding AAV capsid proteins. These ceDNA vectors are structurally distinct from plasmids (including the ceDNA plasmids described herein), which are circular duplex nucleic acid molecules of bacterial origin. Complementary strands of plasmids can be separated after denaturation to produce two nucleic acid molecules, whereas, in contrast, although ceDNA vectors have complementary strands, they are a single DNA molecule and therefore, even if They are denatured, they remain a single molecule. In some embodiments of any of the aspects and embodiments herein, ceDNA vectors can be produced without base methylation of prokaryotic-type DNA, unlike plasmids. Therefore, ceDNA vectors and ceDNA plasmids are different both in terms of structure (in particular, linear compared to circular) and also in view of the methods used to produce and purify these different objects, and furthermore in view of its methylation DNA that is prokaryotic type for ceDNA plasmids and eukaryotic type for the ceDNA vector. Provided herein are capsid-free non-viral ceDNA molecules with covalently closed ends (ceDNA). These non-viral capsid-free ceDNA molecules can be produced in permissive host cells from an expression construct (e.g., a ceDNA plasmid, a ceDNA bacmid, a ceDNA baculovirus, or an integrated cell line) containing a gene. heterologous (e.g., a transgene, in a particular therapeutic transgene) placed between two different inverted terminal repeat (ITR) sequences, where the ITRs are different from each other. In some embodiments of any of the aspects and embodiments herein, one of the ITRs is modified by deletion, insertion and / or substitution compared to a wild-type ITR sequence (e.g., AAV ITR); and at least one of the ITRs comprises a functional terminal resolution site (TRS) and a Rep binding site. The ceDNA vector is preferably duplex, e.g., self-complementary, in at least a portion of the molecule, such as expression cassette (for example, ceDNA is not a double-stranded circular molecule). The ceDNA vector has covalently closed ends and is therefore resistant to exonuclease digestion (e.g. exonuclease I or exonuclease III), for example, for more than one hour at 37°C. In one aspect of any of the aspects or embodiments herein, a ceDNA vector comprises, in the 5' to 3' direction: a first adeno-associated virus (AAV) inverted terminal repeat (ITR), a nucleotide sequence of interest (e.g., an expression cassette as described herein) and a second AAV ITR. In an embodiment of any of the aspects or embodiments herein, the first ITR (ITR 5') and the second ITR (ITR 3') are asymmetrical to each other, that is, they have a 3D spatial configuration different from each other. As an illustrative embodiment, the first ITR may be a wild-type ITR and the second ITR may be a mutated or modified ITR, or vice versa, where the first ITR may be a mutated or modified ITR and the second ITR a wild-type ITR. . In one embodiment of any of the aspects or embodiments herein, the first ITR and the second ITR are both modified but are different sequences, or have different modifications, or are not identical modified ITRs, and have different 3D spatial configurations. In other words, a ceDNA vector with asymmetric ITRs has ITRs where any change in one ITR relative to the WT ITR is not reflected in the other ITR; or alternatively, where the asymmetric ITRs have a pair of modified asymmetric ITRs, they may have a different sequence and a different three-dimensional shape from each other. In one embodiment of any of the aspects or embodiments herein, a ceDNA vector comprises, in the 5' to 3' direction: a first adeno-associated virus (AAV) inverted terminal repeat (ITR), a nucleotide sequence of interest (e.g., an expression cassette as described herein) and a second AAV ITR, wherein the first ITR (5' ITR) and the second ITR (3' ITR) are symmetrical or substantially symmetrical to each other; That is, a ceDNA vector may comprise ITR sequences that have a symmetrical three-dimensional spatial organization such that their structure has the same shape in geometric space, or have the same A, C-C' and B-B' loops in the 3D space. In such an embodiment, a pair of symmetric ITRs or a pair of substantially symmetric ITRs may be modified ITRs (e.g., mod-ITR) that are not wild-type ITRs. A mod-ITR pair may have the same sequence, which has one or more modifications compared to the wild-type ITR and are reverse complements (inverted) to each other. In one embodiment of any of the aspects or embodiments herein, a pair of modified ITRs is substantially symmetrical as defined herein, that is, the pair of modified ITRs may have a different sequence but have the same symmetrical three-dimensional shape or a corresponding one. In some embodiments of any of the aspects and embodiments herein, the symmetric ITRs, or the substantially symmetric ITRs may be wild type (WT ITR) as described herein. That is, both ITRs have a wild-type sequence, but do not necessarily have to be WT ITRs of the same AAV serotype. In one embodiment of any of the aspects or embodiments herein, one WT ITR may be from one AAV serotype, and the other WT ITR may be from a different AAV serotype. In such an embodiment, a WT ITR pair is substantially symmetric as defined herein, that is, it may have one or more conservative nucleotide modifications while retaining symmetrical three-dimensional spatial organization. The wild-type or mutated or otherwise modified ITR sequences provided herein represent DNA sequences included in the expression construct (e.g., ceDNA plasmid, ceDNA bacmid, ceDNA baculovirus) for the production of the vector. ceDNA. Therefore, the ITR sequences actually contained in the ceDNA vector produced from the ceDNA plasmid or other expression construct may or may not be identical to the ITR sequences provided herein as a result of natural changes during the production process. (e.g. replication error). In one embodiment of any of the aspects or embodiments herein, a ceDNA vector described herein comprising the expression cassette with a transgene that is a therapeutic nucleic acid sequence, can be operably linked to one or more regulatory sequences that allow or control the expression of the transgene. In one embodiment of any of the aspects or embodiments herein, the polynucleotide comprises a first ITR sequence and a second ITR sequence, wherein the nucleotide sequence of interest is flanked by the first and second ITR sequences, and the first and second ITR sequences are either asymmetrical to each other, or symmetrical to each other. MA / lz / ¿u¿¿ / uoy 101 In one embodiment of any of the aspects or embodiments herein, an expression cassette is located between two ITRs comprised in the following order with one or more of: a promoter operably linked to a transgene, a post-transcriptional regulatory element and a signal polyadenylation and termination. In one embodiment of any of the aspects or modalities herein, the promoter is adjustable, induced or repressed. The promoter can be any sequence that facilitates transcription of the transgene. In one embodiment of any of the aspects or embodiments herein, the promoter is a CAG promoter, or a variation thereof. The post-transcriptional regulatory element is a sequence that modulates the expression of the transgene, as a non-limiting example, any sequence that creates a tertiary structure that enhances the expression of the transgene, which is a therapeutic nucleic acid sequence. In one embodiment of any of the aspects or embodiments herein, the post-transcriptional regulatory element comprises WPRE. In one embodiment of any of the aspects or embodiments herein, the termination and polyadenylation signal comprises BGHpoIyA. Any cis-regulatory element known in the art, or a combination thereof, may additionally be used, for example, the SV40 late polyA signal upstream enhancer sequence (USE) or other post-transcriptional processing elements including, but not limited to , the thymidine kinase gene of the herpes simplex virus or hepatitis B virus (HBV). In one embodiment of any of the aspects or embodiments herein, the length of the expression cassette in the 5' to 3' direction is greater than the maximum length known to be encapsulated in an AAV shell. In an embodiment of any of the aspects or embodiments herein, the length is greater than 4.6 kb, or greater than 5 kb, or greater than 6 kb, or greater than 7 kb. Various expression cassettes are exemplified herein. In one embodiment of any of the aspects or embodiments herein, the expression cassette may comprise more than 4000 nucleotides, 5000 nucleotides, 10,000 nucleotides or 20,000 nucleotides, or 30,000 nucleotides, or 40,000 nucleotides or 50,000 nucleotides, or any range between about 4000-10,000 nucleotides or 10,000-50,000 nucleotides, or more than 50,000 nucleotides. In one embodiment of any of the aspects or embodiments herein, the expression cassette may further comprise an internal ribosome entry site (IRES) and / or a 2A element. Cis-regulatory elements include, but are not limited to, a promoter, a riboswitch, an insulator, a mir-regulatable element, a post-transcriptional regulatory element, a cell type and tissue-specific promoter, and an enhancer. In some embodiments of any of the aspects and embodiments herein, the ITR may act as a promoter of the transgene. In some embodiments of any of the aspects and embodiments herein, the ceDNA vector comprises additional components for regulating expression of the transgene, for example, a regulatory switch, for controlling and regulating expression of the transgene, and may include, if desired, a regulatory switch that is an inactivation switch to allow controlled cellular inactivation of a cell comprising a ceDNA vector. In one embodiment of any of the aspects or embodiments herein, the ceDNA vectors are capsid-free and can be obtained from a plasmid that encodes in this order: a first ITR, expressed transgene cassette and a second ITR, where at least one of the first and / or second ITR sequences is mutated with respect to the corresponding wild-type AAV2 ITR sequence. In one embodiment of any of the aspects or embodiments herein, the ceDNA vectors described herein are used for therapeutic purposes (e.g., for medical, diagnostic or veterinary uses) or immunogenic polypeptides. The expression cassette may comprise any transgene that is a therapeutic nucleic acid sequence. In certain embodiments, the ceDNA vector comprises any gene of interest in the subject, including one or more polypeptides, peptides, ribozymes, peptide nucleic acids, siRNA, RNAi, antisense oligonucleotides, antisense polynucleotides, antibodies, antigen binding fragments, or any combination of these. In one embodiment of any of the aspects or embodiments herein, the sequences provided in the expression cassette, the expression construct or the donor sequence of a ceDNA vector described herein may have the codons optimized for the host cell. As used herein, the term undergo codon optimization or codon optimization refers to the process of modifying a nucleic acid sequence to improve expression in cells of the vertebrate of interest, for example, mouse or human, by replacing at least one, more than one, or a significant number of codons from the native sequence (e.g., a prokaryotic sequence) with codons that are used most or most frequently in the genes of that vertebrate. Various species exhibit a particular bias for certain codons of a particular amino acid. Typically, codon optimization does not alter the amino acid sequence of the original translated protein. Optimized codons can be determined using, for example, Aptagen's Gene Forge® custom gene synthesis and codon optimization platform (Aptagen, Inc., 2190 Fox Mili Rd. Suite 300, Herndon, Va. 20171) or another database. publicly available data. Many organisms show a bias toward using particular codons to encode the insertion of a particular amino acid into a growing peptide chain. Codon preference or codon bias, differences in codon usage between organisms, is achieved through degeneration of the genetic code, and this is documented among many organisms. Codon bias is often correlated with the efficiency of messenger RNA (mRNA) translation, which in turn is thought to depend on, among others, the properties of the codons being translated. ΜΛ / t / zuzz / uoy l OI and the availability of particular transfer RNA (tRNA) molecules. The predominance of selected tRNAs in a cell is generally a reflection of the codons most frequently used in peptide synthesis. Consequently, genes can be tailored for optimal gene expression in a given organism based on codon optimization. Given the large number of gene sequences available for a wide variety of animal, plant and microbial species, it is possible to calculate the relative frequencies of codon usage (Nakamura, Y., et al., Codon usage tabulated from the international DNA sequence databases: status forthe year 2000 Nucí. Acids Res. 28:292 (2000)). Inverted Terminal Repeats (ITR) As described herein, ceDNA vectors are capsid-free linear duplex DNA molecules formed from a continuous strand of complementary DNA with covalently closed ends (linear, continuous and non-encapsulated structure), comprising a sequence of 5' inverted terminal repeat (ITR) and a 3' ITR sequence that are different or asymmetric from each other. At least one of the ITRs comprises a terminal resolution site and a replication protein binding site (RPS) (sometimes called a replication protein binding site), for example, a Rep binding site. Generally, The ceDNA vector contains at least one modified AAV inverted terminal repeat (ITR) sequence, that is, a deletion, insertion and / or substitution with respect to the other ITR, and a transgene expressing it. In one embodiment of any of the aspects or embodiments herein, at least one of the ITRs is an AAV ITR, for example, a wild-type AAV ITR. In one embodiment of any of the aspects or embodiments herein, at least one of the ITRs is a modified ITR with respect to the other ITR, that is, the ceDNA comprises ITRs that are asymmetrical with respect to each other. In an embodiment of any of the aspects or embodiments herein, at least one of the ITRs is a non-functional ITR. In one embodiment of any of the aspects or embodiments herein, the ceDNA vector comprises: (1) an expression cassette comprising a cis-regulatory element, a promoter and at least one transgene; or (2) a promoter operatively linked to at least one transgene, and (3) two self-complementary sequences, e.g., ITR flanking said expression cassette, wherein the ceDNA vector is not associated with a capsid protein. In some embodiments of any of the aspects and embodiments herein, the ceDNA vector comprises two self-complementary sequences found in an AAV genome, where at least one comprises an operational Rep-binding element (RBE) and a binding site. terminal resolution (TRS) of AAV or a functional variant of the RBE, and one or more cis-regulatory elements operatively linked to a transgene. In some embodiments of any of the aspects and embodiments herein, the ceDNA vector comprises additional components to regulate the expression of the transgene, for example, regulatory switches, to control and regulate the expression of the transgene. MA / t / zuzz / uoy / 01 expression of the transgene, and may include a regulatory switch that is an inactivation switch to allow controlled cellular inactivation of a cell comprising a ceDNA vector. In one embodiment of any of the aspects or embodiments herein, the two self-complementary sequences may be ITR sequences from any known parvovirus, for example, a dependovirus such as AAV (for example, AAV1-AAV12). Any AAV serotype can be used, including, but not limited to, a modified AAV2 ITR sequence, which retains a Rep binding site (RBS) such as 5-GCGCGCTCGCTCGCTC-3' and a terminal resolution site (TRS). ), in addition to a variable palindromic sequence that allows the formation of hairpin secondary structures. In some embodiments of any of the aspects and embodiments herein, an ITR may be synthetic. In one embodiment of any of the aspects or embodiments herein, a synthetic ITR is based on ITR sequences from more than one AAV serotype. In another embodiment, a synthetic ITR does not include any AAV sequences. In yet another embodiment, a synthetic ITR retains the ITR structure described above, although it has only some AAV source sequences or none at all. In some aspects, a synthetic ITR may preferentially interact with a wild-type Rep or a Rep of a specific serotype, or in some cases it will not be recognized by a wild-type Rep and will be recognized only by a mutated Rep. In some embodiments of any of the aspects or embodiments herein, the ITR is a synthetic ITR sequence that retains a functional Rep binding site (RBS) such as 5'-GCGCGCTCGCTCGCTC-3' and a terminal resolution site (TRS). ) in addition to a variable palindromic sequence that allows the formation of a hairpin secondary structure. In some examples, a modified ITR sequence retains the RBS, TRS sequence and the structure and position of a Rep binding element that forms the terminal loop portion of one of the ITR hairpin secondary structures of the corresponding sequence of the ITR of wild-type AAV2. Illustrative ITR sequences for use in ceDNA vectors are described in Tables 2-9, 10A and 10B, SEQ ID NO: 2, 52, 101-449 and 545-547, and partial ITR sequences are described in shown in Figures 26A-26B of PCT application no. PCT / US 18 / 49996, filed September 7, 2018. In some embodiments of any of the aspects or embodiments herein, a ceDNA vector may comprise an ITR with a modification in the ITR corresponding to any of the modifications in the ITR sequences or partial ITR sequences shown in one or more of Tables 2, 3, 4, 5, 6, 7, 8, 9, 10A and 10B, PCT application no. PCT / US 18 / 49996, filed September 7, 2018. In one embodiment of any of the aspects or embodiments herein, ceDNA vectors can be produced from expression constructs that further comprise a specific combination of cis-regulatory elements. Cis-regulatory elements include, but are not limited to, a promoter, a riboswitch, an insulator, a mir-regulatable element, a post-transcriptional regulatory element, a cell type and tissue-specific promoter, and an enhancer. In some embodiments of any of the aspects and embodiments herein, the ITR may act as a promoter of the transgene. In some embodiments of any of the aspects and embodiments herein, the ceDNA vector comprises additional components for regulating expression of the transgene, for example, regulatory switches as described in PCT application no. PCT / US 18 / 49996, filed September 7, 2018, to regulate the expression of the transgene, or an inactivation switch, that can inactivate a cell comprising the ceDNA vector. In one embodiment of any of the aspects or embodiments herein, the expression cassettes may further include a post-transcriptional element to increase expression of a transgene. In one embodiment of any of the aspects or embodiments herein, the woodchuck hepatitis virus (WHP) post-transcriptional regulatory element (WPRE) is used to increase expression of a transgene. Other post-transcriptional processing elements may be used, such as the post-transcriptional element of the thymidine kinase gene of herpes simplex virus or hepatitis B virus (HBV). Secretory sequences can be linked to transgenes, for example, VH-02 and VK-A26 sequences. The expression cassettes may include a polyadenylation sequence known in the art or a variation thereof, such as a natural sequence isolated from bovine BGHpA or an SV40pA virus, or a synthetic sequence. Some expression cassettes may further include the enhancer sequence upstream of the SV40 late polyA signal (USE). EUS can be used in combination with SV40pA or heterologous poly-A signal. Figures 1A-1C of international application no. PCT / US2018 / 050042, filed on September 7, 2018 and incorporated herein by reference in its entirety, show schemes of illustrative non-limiting ceDNA vectors, or the corresponding sequence of ceDNA plasmids. ceDNA vectors do not have a capsid and can be obtained from a plasmid that encodes in this order: a first ITR, expressible transgene cassette and a second ITR, where at least one of the first and / or second ITR sequences is mutated. with respect to the corresponding wild-type AAV2 ITR sequence. The expressible transgene cassette preferably includes one or more of, in this order: an enhancer / promoter, an ORF reporter (transgene), a post-transcriptional regulatory element (e.g., WPRE), and a polyadenylation and termination signal (e.g. , BGH polyA). Promoters Suitable promoters, including those described above, may be derived from viruses and therefore may be referred to as viral promoters, or may be derived from any organism, including prokaryote or eukaryotic organisms. Suitable promoters can be used to induce expression by any RNA polymerase (eg, pol I, pol II, pol III). Illustrative promoters include, but are not limited to, the SV40 early promoter, the mouse mammary tumor virus long terminal repeat (LTR) promoter; late promoter MA / IZ / ¿U¿¿ / UO3 / ΟΊ main adenovirus (Ad MLP); a herpes simplex virus (HSV) promoter, a cytomegalovirus (CMV) promoter, such as the CMV immediate early promoter region (CMVTE), a Rous sarcoma virus (RSV) promoter, a human U6 small nuclear promoter (U6, for example, (Miyagishi et al., Nature Biotechnology 20, 497-500 (2002)), an improved U6 promoter (e.g., Xia et al., Nucleic Acids Res., September 1, 2003; 31( 17)), a human H1 promoter (H1), a CAG promoter, a human alpha l-antitypsin (HAAT) promoter (for example and the like). In one embodiment of any of the aspects or embodiments herein, these promoters are modified at their downstream intron-containing end to include one or more nuclease cleavage sites. In one embodiment of any of the aspects or embodiments herein, the DNA containing the nuclease cleavage site(s) It is foreign to the promoter DNA. In one embodiment of any of the aspects or embodiments herein, a promoter may comprise one or more transcription-specific regulatory sequences to further enhance expression and / or modify the spatial expression and / or temporal expression thereof. A promoter may further comprise distal enhancer or repressor elements, which may be located up to several thousand base pairs from the transcription start site. A promoter can be derived from sources including viruses, bacteria, fungi, plants, insects and animals. A promoter may regulate the expression of a gene component constitutively or differentially with respect to the cell, tissue or organ in which the expression occurs, or with respect to the developmental stage in which the expression occurs, or in response to external stimuli such as physiological stresses, pathogens, metal ions or inducing agents. Representative examples of promoters include bacteriophage T7 promoter, bacteriophage T3 promoter, SP6 promoter, lac operator promoter, tac promoter, SV40 late promoter, SV40 early promoter, RSV-LTR promoter, CMV promoter IE, the SV40 early promoter or the SV40 late promoter, and the CMV IE promoter, as well as the promoters listed below. Such promoters and / or enhancers can be used for the expression of any gene of interest, for example, therapeutic proteins). For example, the vector may comprise a promoter that is operably linked to the nucleic acid sequence encoding a therapeutic protein. In one embodiment of any of the aspects or embodiments herein, the promoter operably linked to the coding sequence of the therapeutic protein may be a simian virus 40 (SV40) promoter, a mouse mammary tumor virus (MMTV) promoter, or a mouse mammary tumor virus (MMTV) promoter. , a human immunodeficiency virus (HIV) promoter such as the bovine immunodeficiency virus (BIV) long terminal repeat (LTR) promoter, a Moloney virus promoter, an avian leukemia virus (ALV) promoter ), a cytomegalovirus (CMV) promoter such as the CMV immediate early promoter, the Epstein-Barr virus (EBV) promoter, or a Rous sarcoma virus (RSV) promoter. In one embodiment of any of the aspects or embodiments herein, the promoter may further be a promoter of a human gene such as human ubiquitin C (hUbC), human actin, human myosin, human hemoglobin, human muscle creatine or human metallothionein. The promoter may further be a tissue-specific promoter, such as a liver-specific promoter, such as human alpha l-antitypsin (HAAT) or transthyretin (TTR), natural or synthetic. In one embodiment of any of the aspects or embodiments herein, administration to the liver can be achieved through the use of specific targeting of endogenous ApoE of the composition comprising a ceDNA vector, to hepatocytes through the lipoprotein receptor of low density (LDL) present on the surface of the hepatocyte. In one embodiment of any of the aspects or embodiments herein, the promoter used is the native promoter of the gene encoding the therapeutic protein. The promoters and other regulatory sequences for the respective genes encoding the therapeutic proteins are known and have been characterized. The promoter region used may further include one or more additional (e.g., native) regulatory sequences, such as enhancers (e.g., the Serpin enhancer) known in the art. Non-limiting examples of promoters suitable for use in accordance with the present invention include the CAG promoter of, for example, the HAAT promoter, the human EF1 -a promoter or a fragment of the EF1 -a promoter and the rat EF1-a promoter. Polyadenylation sequences A sequence encoding a polyadenylation sequence may be included in the ceDNA vector to stabilize the mRNA expressed from the ceDNA vector and to assist in nuclear export and translation. In one embodiment of any of the aspects or embodiments herein, the ceDNA vector does not include a polyadenylation sequence. In other embodiments, the vector includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40 , at least 45, at least 50 or more adenine dinucleotides. In some embodiments of any of the aspects or embodiments herein, the polyadenylation sequence comprises about 43 nucleotides, about 40-50 nucleotides, about 40-55 nucleotides, about 45-50 nucleotides, about 35-50 nucleotides, or any range. between these. In one embodiment of any of the aspects or embodiments herein, the ceDNA can be obtained from a vector polynucleotide encoding a heterologous nucleic acid operably positioned between two different inverted terminal repeat (ITR) sequences (e.g., AAV ITR), wherein at least one of the ITRs comprises a terminal resolution site and a replicative protein binding site (RPS), e.g., a Rep binding site (e.g., wt AAV ITR), and one of the ITRs comprises a delete, insertion and / or substitution with respect to the other ITR, for example, functional ITR. MA / IZ / ZUZZ / UOU l OI In one embodiment of any of the aspects or embodiments herein, the host cells do not express viral capsid proteins and the polynucleotide vector template lacks any viral capsid coding sequence. In one embodiment of any of the aspects or embodiments herein, the polynucleotide vector template lacks AAV capsid genes but also capsid genes from other viruses). In one embodiment of any of the aspects or embodiments herein, the nucleic acid molecule further lacks coding sequences for the AAV Rep protein. Accordingly, in some embodiments of any of the aspects and embodiments herein, the nucleic acid molecule of the invention lacks functional genes for both AAV cap and AAV rep. In one embodiment of any of the aspects or embodiments herein, the ceDNA vector does not have a modified ITR. In one embodiment of any of the aspects or embodiments herein, the ceDNA vector comprises a regulatory switch as described herein (or in PCT Application No. PCT / US 18 / 49996, filed September 7, 2018 ). IV. PRODUCTION OF A ceDNA VECTOR Methods for the production of a ceDNA vector as described herein comprising a pair of asymmetric ITRs or a pair of symmetric ITRs, as defined herein, are described in section IV of PCT / US 18 / 49996 filed on September 7, 2018, which is incorporated herein by reference in its entirety. As described herein, the ceDNA vector can be obtained, for example, by the process comprising the steps of: a) incubating a population of host cells (e.g., insect cells) harboring the template of the expression construct of the polynucleotide (for example, a ceDNA plasmid, a ceDNA bacmid and / or a ceDNA baculovirus), lacking viral capsid coding sequences, in the presence of a Rep protein under effective conditions and for a time sufficient to induce producing the ceDNA vector within the host cells, and wherein the host cells do not comprise viral capsid coding sequences; and b) harvest and isolate the ceDNA vector from the host cells. The presence of Rep protein induces replication of the vector polynucleotide with a modified ITR to produce the ceDNA vector in a host cell. However, viral particles (e.g., AAV virions) are not expressed. Therefore, there is no size limitation, such as that naturally imposed on AAV or other virus-based vectors. The presence of the ceDNA vector isolated from the host cells can be confirmed by digestion of the DNA isolated from the host cell with a restriction enzyme that has a unique recognition site on the ceDNA vector and by analyzing the digested DNA material in a non-denaturing gel to confirm the presence of bands characteristic of linear and continuous DNA compared to linear and non-continuous DNA. In one embodiment of any of the aspects or embodiments herein, the invention provides the use of host cell lines that have stably integrated the DNA vector polynucleotide expression template (ceDNA template) into their own genome in the production of the non-viral DNA vector, for example, as described in Lee, L. et al., (2013) Píos One 8(8): e69879. Preferably, Rep is added to the host cells at an MOI of about 3. When the host cell line is a mammalian cell line, for example, HEK293 cells, the cell lines may have a stably integrated polynucleotide vector template, and a second vector, such as herpes virus, can be used to introduce the Rep protein into cells, allowing cleavage and amplification of ceDNA in the presence of Rep and helper virus. In one embodiment of any of the aspects or embodiments herein, the host cells used to prepare the ceDNA vectors described herein are insect cells, and the baculovirus is used to deliver both the polynucleotide encoding the Rep protein and the Non-viral DNA vector polynucleotide expression construct template for ceDNA. In some embodiments of any of the aspects or embodiments herein, the host cell is modified to express the Rep protein. The ceDNA vector is then harvested and isolated from the host cells. The time for harvesting the ceDNA vectors described herein from cells can be selected and optimized to achieve high-throughput production of the ceDNA vectors. For example, the collection time can be selected in view of cell viability, cell morphology, cell growth, etc. In one embodiment of any of the aspects or embodiments herein, the cells are cultured under sufficient conditions and harvested for a sufficient time after baculoviral infection to produce ceDNA vectors, but before the majority of the cells begin to die due to baculoviral toxicity. DNA vectors can be isolated using plasmid purification kits, such as Qlagen Endo-Free Plasmid kits. Other methods developed for plasmid isolation can be adapted for DNA vectors. Generally, any nucleic acid purification method can be adopted. The DNA vectors can be purified by any means known to those skilled in the art for DNA purification. In one embodiment of any of the aspects or embodiments herein, the ceDNA vectors are purified as DNA molecules. In one embodiment of any of the aspects or embodiments herein, the ceDNA vectors are purified as exosomes or microparticles. The presence of the ceDNA vector can be confirmed by digestion of the vector DNA isolated from the cells with a restriction enzyme that has a unique recognition site on the vector DNA and by analyzing both the DNA material MA / t / zuzz / uoy 101 digested and undigested through gel electrophoresis to confirm the presence of characteristic bands of linear and continuous DNA compared to linear and non-continuous DNA. V. PREPARATION OF LIPID PARTICLES Lipid particles (e.g., lipid nanoparticles) can form spontaneously upon mixing of TNA (e.g., ceDNA) and lipids. Depending on the desired particle size distribution, the resulting nanoparticle mixture can be extruded through a membrane (e.g. 100 nm limit) by using, for example, a thermobarrel extruder, such as Lipex Extruder ( Northern Lipids, Inc). In some cases, the extrusion step can be omitted. Ethanol removal and simultaneous buffer exchange can be performed, for example, by dialysis or tangential flow filtration. Generally, lipid particles (e.g., lipid nanoparticles) can be formed by any method known in the art. For example, lipid particles (e.g., lipid nanoparticles) can be prepared by the methods described, for example, in US2013 / 0037977, US2010 / 0015218, US2013 / 0156845, US2013 / 0164400, US2012 / 0225129 and US2010 / 0130588, the contents of which are incorporated herein by reference in their entirety. In some embodiments of any of the aspects or embodiments herein, lipid particles (e.g., lipid nanoparticles) may be prepared by using a continuous mixing method, a direct dilution process, or an online dilution process. Processes and apparatus for preparing lipid nanoparticles by using direct dilution and online dilution processes are described in US2007 / 0042031, the contents of which are incorporated herein by reference in their entirety. Processes and apparatus for preparing lipid nanoparticles by using stepwise dilution processes are described in US2004 / 0142025, the contents of which are incorporated herein by reference in their entirety. In one embodiment of any of the aspects or embodiments herein, lipid particles (e.g., lipid nanoparticles) can be prepared by an impact jet process. Generally, the particles are formed by mixing lipids dissolved in alcohol (for example, ethanol) with ceDNA dissolved in a buffer, for example, a citrate buffer, a sodium acetate and magnesium chloride buffer, a malic acid buffer, a malic acid and sodium chloride buffer, or a sodium citrate and sodium chloride buffer. The mixing ratio of lipids to ceDNA can be about 45-55% lipid and about 6545% ceDNA. The lipid solution may contain an ionizable lipid, a non-cationic lipid (for example, a phospholipid, such as DSPC, DOPE and DOPC), PEG or PEG-conjugated molecule (for example, PEG-lipid) and a ester! (e.g. cholesterol) at a total lipid concentration of 5 to 30 MA / t / zuzz / uoy1 OI mg / ml, most likely 5 to 15 mg / mL, most likely 9 to 12 mg / mL in an alcohol, for example, in ethanol. In the lipid solution, the molar ratio of the lipids can vary from about 25-98% for the cationic lipid, preferably from about 35-65%; about 0-15% for the nonionic lipid, preferably about 0-12%; about 0-15% for the PEG or PEG-conjugated lipid molecule, preferably about 1-6%; and about 0-75% for the ester, preferably about 30-50%. The ceDNA solution may comprise ceDNA in a concentration range of 0.3 to 1.0 mg / mL, preferably 0.3-0.9 mg / mL in buffered solution, with a pH in the range of 3.5-5. To form the LNPs, in an illustrative but non-limiting embodiment, the two liquids are heated to a temperature in the range of about 15-40°C, preferably about 30-40°C, and then mixed, for example, in a impact jet mixer, which instantly forms the LNP. The mixing flow rate can vary from 10-600 mL / min. The tube ID can have a range of 0.25 to 1.0 mm and a total flow rate of 10-600 mUrnin. The combination of flow regime and tube ID can have the effect of controlling the particle size of LNPs between 30 and 200 nm. The solution may then be mixed with a buffered solution at a higher pH with a mixing ratio in the range of 1:1 to 1:3 volLvol, preferably about 1:2 volLvol. If necessary, this buffer solution can be at a temperature in the range of 15-40 °C or 30-40 °C. The mixed LNPs can then be subjected to an anion exchange filtration step. Before anion exchange, the mixed LNPs can be incubated for a period of time, for example, 30 minutes to 2 hours. The temperature during incubation can be in the range of 15-40 °C or 30-40 °C. After incubation, the solution is filtered through a filter, such as a 0.8 pm filter, which contains an anion exchange separation step. This process can use tube IDs ranging from 1 mm to 5 mm and a flow rate of 10 to 2000 mL / min. After formation, the LNPs can be concentrated and diafiltered by an ultrafiltration process where the alcohol is removed and the buffer is exchanged for the final buffer solution, for example, phosphate buffered saline (PBS) at a pH of approximately 7, for example, a pH of about 6.9, a pH of about 7.0, a pH of about 7.1, a pH of about 7.2, a pH of about 7.3, or a pH of about 7.4. The ultrafiltration process can use a tangential flow filtration (TFF) format by using a nominal membrane molecular weight cutoff range of 30-500 kD. The membrane format is hollow fiber or flat sheet cassette. TFF processes with the appropriate molecular weight limit can retain the LNP in the retentate and the filtrate or permeate contains the alcohol; citrate buffer and residues of the final buffer. The TFF process is a multi-step process with an initial concentration at a ceDNA concentration of 1-3 mg / mL. After concentration, the LNP solution is diafiltered against the final buffer for 10 to 20 volumes to remove alcohol and perform buffer exchange. The material can then be concentrated 1 to 3 times more. The concentrated LNP solution can be sterilized by filtration. ma / t / zuzz / uoy l OI SAW. PHARMACEUTICAL COMPOSITIONS AND FORMULATIONS Further provided herein is a pharmaceutical composition comprising the TNA lipid particle and a pharmaceutically acceptable carrier or excipient. In one embodiment of any of the aspects or embodiments herein, the TNA lipid particles (e.g., lipid nanoparticles) are provided with full encapsulation, partial encapsulation of the therapeutic nucleic acid. In one embodiment of any of the aspects or embodiments herein, the nucleic acid therapeutic agent is completely encapsulated in the lipid particles (e.g., lipid nanoparticles) to form a nucleic acid-containing lipid particle. In one embodiment of any of the aspects or embodiments herein, the nucleic acid may be encapsulated within the lipid portion of the particle, thereby protecting it from enzymatic degradation. In one embodiment of any of the aspects or embodiments herein, the lipid particle has an average diameter of about 20 nm to about 100 nm, from 30 nm to about about about about about about 150nm,nm 130nm,nm 100 nm, about to about about to about about about nm 150nm,nm 100 nm, nm to about 150 nm, about 60 nm to about 110 nm, about 80 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, about 70 nm to about 80 nm, or about 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm, 75nm, 80nm, 85nm, 90nm, 95nm, 100nm, 105nm nm, 110nm, 115nm, 120nm, 125nm, 130nm, 135nm, 140nm, 145nm or 150nm to ensure effective delivery. Nucleic acid-containing lipid particles (e.g., lipid nanoparticles) and their preparation method are described, for example, in PCT / US18 / 50042, United States Patent Publications No. 20040142025 and 20070042031, the descriptions of which are incorporated herein by reference in their entirety for all purposes. In one embodiment of any of the aspects or embodiments herein, the size of lipid particle (e.g., lipid nanoparticle) can be determined by quasi-elastic light scattering using, for example, a Malvern Zetasizer Nano ZS system (Malvem, United Kingdom). . Generally, the lipid particles (e.g., lipid nanoparticles) of the invention have a mean diameter selected to provide a desired therapeutic effect. Depending on the intended use of the lipid particles (e.g., lipid nanoparticles), the proportions of the components may vary and the delivery efficiency of a particular formulation may be measured through, for example, an endosomal release parameter assay ( ERP). In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) may be conjugated with other moieties to prevent aggregation. Such lipid conjugates include, but are not limited to, PEG-lipid conjugates such as, for example, PEG coupled to dialkyloxypropyls (e.g., PEGDAA conjugates), PEG coupled to diacylglycerols (e.g., PEG-DAG conjugates), PEG coupled to cholesterol, PEG coupled to phosphatidylethanolamines, and PEG conjugated to ceramides (see, for example, US Patent 5,885,613), cationic PEG lipids, polyoxazoline (POZ)-lipid conjugates (for example, POZ-lipid conjugates DAA; see, for example, US Provisional Application 61 / 294,828, filed January 13, 2010 and US Provisional Application No. 61 / 295,140, ​​filed January 14, 2010), polyamide oligomers ( for example, ATTA-lipid conjugates) and mixtures thereof. In PCT publication no. Additional examples of POZ-lipid conjugates are described in WO 2010 / 006282. PEG or POZ can be conjugated directly to the lipid or can be attached to the lipid through a linker moiety. Any suitable linker moiety can be used to couple PEG or POZ to a lipid, including, for example, non-ester containing linker moieties and ester containing linker moieties. In certain preferred embodiments, non-ester containing linking moieties, such as amides or carbamates, are used. The descriptions of each of the foregoing patent documents are incorporated herein by reference in their entirety for all purposes. In one embodiment of any of the aspects or embodiments herein, the ceDNA may form a complex with the lipid portion of the particle or be encapsulated in the lipid position of the lipid particle (e.g., lipid nanoparticle). In one embodiment of any of the aspects or embodiments herein, the ceDNA can be completely encapsulated in the lipid position of the lipid particle (e.g., lipid nanoparticle), which protects it from degradation by a nuclease, for example, in an aqueous solution. In one embodiment of any of the aspects or embodiments herein, the ceDNA in the lipid particle (e.g., lipid nanoparticle) is not substantially degraded after exposure of the lipid particle (e.g., lipid nanoparticle) to a nuclease. 37 °C for at least approximately 20, 30, 45 or 60 minutes. In some embodiments of any of the aspects or embodiments herein, the ceDNA in the lipid particle (e.g., lipid nanoparticle) is not substantially degraded after incubation of the particle in serum at 37 ° C for at least about 30, 45, or 60 minutes or at least approximately 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34 or 36 hours. In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) are substantially non-toxic to a subject, for example, to a mammal, such as a human. In one embodiment of any of the aspects or embodiments herein, a pharmaceutical composition comprising a therapeutic nucleic acid of the present disclosure may be formulated into lipid particles (e.g., lipid nanoparticles). In some embodiments of any of the aspects and embodiments herein, the lipid particle comprising a therapeutic nucleic acid may be formed from a described ionizable lipid. In some other embodiments, the lipid particle comprising a therapeutic nucleic acid may be formed from a non-cationic lipid. In a preferred embodiment, the lipid particle of the invention is a nucleic acid-containing lipid particle, which is formed from an ionizable lipid comprising a therapeutic nucleic acid selected from the group consisting of mRNA, antisense RNA and oligonucleotide, ribozymes, aptamer, RNA interference (RNAi), Dicer substrate dsRNA, small hairpin RNA (shRNA), asymmetric RNA interference (aRNA), microRNA (miRNA), minicircle DNA, minigene, viral DNA (e.g. AAV genome or lentivirus) or non-viral synthetic DNA vectors, closed-end linear duplex DNA (ceDNA / CELID), plasmids, bacmids, doggybone™ DNA vectors, minimalist immunologically defined gene expression (MIDGE) vector, ministrand DNA vector non-viral (covalently closed linear DNA vector) or minimal dumbbell-shaped DNA vector (dumbbell DNA). In another preferred embodiment, the lipid particle of the invention is a nucleic acid-containing lipid particle, which is formed from a non-cationic lipid and optionally a conjugated lipid that prevents aggregation of the particle. In one embodiment of any of the aspects or embodiments herein, the lipid particle formulation is an aqueous solution. In one embodiment of any of the aspects or embodiments herein, the lipid particle formulation (e.g., lipid nanoparticles) is a lyophilized powder. According to some aspects, the disclosure provides a lipid particle formulation further comprising one or more pharmaceutical excipients. In one embodiment of any of the aspects or embodiments herein, the lipid particle formulation (e.g., lipid nanoparticle) further comprises sucrose, tris, trehalose and / or glycine. In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) described herein may be incorporated into pharmaceutical compositions suitable for administration to a subject for in vivo delivery to cells, tissues or organs of the subject. Typically, the pharmaceutical composition comprises the lipid particles (e.g., lipid nanoparticles) of TNA described herein and a pharmaceutically acceptable carrier. In one embodiment of any of the aspects or embodiments herein, the TNA lipid particles (e.g., lipid nanoparticles) of the disclosure may be incorporated into a pharmaceutical composition suitable for a desired route of therapeutic administration (e.g., parenteral administration). . Passive transduction of tissues by high-pressure intravenous or intra-arterial infusion, as well as intracellular injection, such as intranuclear microinjection or intracytoplasmic injection, is also contemplated. Pharmaceutical compositions for therapeutic purposes may be formulated as a solution, microemulsion, dispersion, liposome or other ordered structure suitable for a high concentration of ceDNA vector. Sterile injectable solutions can be prepared by incorporating the ceDNA vector compound in the required amount in a suitable buffer with one of the ingredients listed above, or a combination of these, as required, followed by filtered sterilization. A lipid particle as described herein may be incorporated into a pharmaceutical composition suitable for topical, systemic, intraamniotic, intrathecal, intracranial, intraarterial, intravenous, intralymphatic, intraperitoneal, subcutaneous, tracheal, intratissue (e.g., intramuscular, intracardiac, intrahepatic, intrarenal, intracerebral), intrathecal, intravesical, conjunctival (e.g., extraorbital, intraorbital, retroorbital, intraretinal, subretinal, choroidal, subchoroidal, intrastromal, intracameral, and intravitreal), intracochlear, and mucosa (e.g., oral, rectal, nasal) . Passive transduction of tissues by high-pressure intravenous or intra-arterial infusion, as well as intracellular injection, such as intranuclear microinjection or intracytoplasmic injection, is also contemplated. Pharmaceutically active compositions comprising TNA lipid particles (e.g., lipid nanoparticles) can be formulated to deliver a transgene in the nucleic acid to the cells of a recipient, resulting in therapeutic expression of the transgene therein. The composition may further include a pharmaceutically acceptable carrier. Typically, pharmaceutical compositions for therapeutic purposes must be sterile and stable under the conditions of manufacture and storage. The composition may be formulated as a solution, microemulsion, dispersion, liposome or other ordered structure suitable for high ceDNA vector concentration. Sterile injectable solutions can be prepared by incorporating the ceDNA vector compound in the required amount in a suitable buffer with one of the ingredients listed above, or a combination of these, as required, followed by filtered sterilization. In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) are solid core particles that possess at least one lipid bilayer. In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) have a non-bilayer structure, that is, a non-lamellar (i.e., non-bilayer) morphology. Without limitations, non-bilayer morphology may include, for example, three-dimensional tubes, rods, cubic symmetries, etc. The non-lamellar morphology (i.e., non-bilayer structure) of lipid particles (e.g., lipid nanoparticles) can be determined by analytical techniques known and used by those skilled in the art. Such techniques include, but are not limited to, cryogenic transmission electron microscopy (Cryo-TEM), differential scanning calorimetry (DSC), X-ray diffraction and the like. For example, the morphology of lipid particles (lamellar versus non-lamellar) can be easily assessed and characterized by, for example, Cryo-TEM analysis as described in US2010 / 0130588, the contents of which are incorporated herein. for reference in its entirety. In one embodiment of any of the aspects or embodiments herein, the lipid particles (e.g., lipid nanoparticles) have a non-lamellar morphology and are electron dense. In one embodiment of any of the aspects or embodiments herein, the disclosure provides a lipid particle (e.g., lipid nanoparticle) having a unilamellar or multilamellar structure. In some aspects, the disclosure provides a formulation of lipid particles (e.g., lipid nanoparticles) comprising multivesicular particles and / or foam-based particles. By controlling the composition and concentration of the lipid components, one can control the rate at which the lipid conjugate is exchanged from the lipid particle and, in turn, the rate at which the lipid particle (e.g., lipid nanoparticle) it becomes fusogenic. Additionally, other variables, including, for example, pH, temperature, or ionic strength, may be used to vary and / or control the rate at which the lipid particle (e.g., lipid nanoparticle) becomes fusogenic. Other methods that can be used to control the rate at which the lipid particle (e.g., lipid nanoparticle) becomes fusogenic will be apparent to those skilled in the art based on this disclosure. It will further be evident that, by controlling the composition and concentration of the lipid conjugate, the lipid particle size can be controlled. In one embodiment of any of the aspects or embodiments herein, the pKa of the formulated ionizable lipids can be correlated with the effectiveness of the LNPs for delivery of nucleic acids (see Jayaraman et al., Angewandte Chemie, international edition (2012) , 51 (34), 85298533; Semple et al., Nature Biotechnology 28, 172-176 (2010), both incorporated by reference in their entirety). In one embodiment of any of the aspects or embodiments herein, the preferred pKa range is about 5 to about 8. In one embodiment of any of the aspects or embodiments herein, the preferred pKa range is about 6. to about 7. In one embodiment of any of the aspects or embodiments herein, the preferred pKa is about 6.5. In one embodiment of any of the aspects or embodiments herein, the pKa of the ionizable lipid can be determined in lipid particles (e.g., lipid nanoparticles) by using a 2-(p-toluidino acid fluorescence-based assay. )-6-naphthalenesulfoníco (TNS). In one embodiment of any of the aspects or embodiments herein, the encapsulation of ceDNA in lipid particles (e.g., lipid nanoparticles) can be determined by the embodiment of a membrane-impermeable fluorescent dye exclusion assay, which uses a dye which enhances fluorescence when associated with nucleic acid, for example, an OliGreen® assay or PicoGreen® assay. Generally, encapsulation is determined by adding the dye to the lipid particle formulation, measuring the resulting fluorescence, and comparing it to the fluorescence observed after the addition of a small amount of nonionic detergent. Detergent-mediated disruption of the lipid bilayer releases the encapsulated ceDNA, allowing it to interact with the membrane-impermeable dye. The encapsulation of ceDNA can be calculated as E= (lo - l) / lo, where I and lo refer to the fluorescence intensities before and after the addition of detergent. Unit dose In one embodiment of any of the aspects or embodiments herein, the pharmaceutical compositions may be presented in unit dosage form. A unit dosage form will typically be adapted to one or more specific routes of administration of the pharmaceutical composition. In some embodiments of any of the aspects and embodiments herein, the unit dosage form is adapted for administration by inhalation. In some embodiments of any of the aspects and embodiments herein, the unit dosage form is adapted for administration by a vaporizer. In some embodiments of any of the aspects and embodiments herein, the unit dosage form is adapted for administration by a nebulizer. In some embodiments of any of the aspects and embodiments herein, the unit dosage form is adapted for administration by means of an aerosol. In some embodiments of any of the aspects and embodiments herein, the unit dosage form is adapted for oral administration, for buccal administration, or for sublingual administration. In some embodiments of any of the aspects or embodiments herein, the unit dosage form is adapted for intravenous, intramuscular or subcutaneous administration. In some embodiments of any of the aspects and embodiments herein, the unit dosage form is adapted for intrathecal or intracerebroventricular administration. In some embodiments of any of the aspects and embodiments herein, the pharmaceutical composition is formulated for topical administration. The amount of active ingredient that can be combined with a carrier material to ΜΛ / t / zuzz / uoy 101 producing a single dosage form will generally be the amount of the compound that produces a therapeutic effect. Vile. TREATMENT METHODS The ionizable lipid composition and methods (e.g., lipid particles (e.g., lipid nanoparticles) of TNA as described herein) described herein can be used to introduce a nucleic acid sequence (e.g., a sequence of therapeutic nucleic acids) in a host cell. In one embodiment of any of the aspects or embodiments herein, the introduction of a nucleic acid sequence into a host cell through the use of the LNPs with TNA (for example, lipid particles (for example, lipid nanoparticles) of the vector ceDNA as described herein) can be monitored with appropriate biomarkers from treated patients to assess gene expression. The LNP compositions provided herein can be used to deliver a transgene (a nucleic acid sequence) for various purposes. In one embodiment of any of the aspects or embodiments herein, the ceDNA vectors (e.g., the lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein) can be used in various ways, which include, for example, ex situ, in vitro and in vivo applications, methodologies, diagnostic procedures and / or gene treatment regimens. Provided herein are methods of treating a disease or disorder in a subject comprising introducing into a target cell in need (e.g., a liver cell, a muscle cell, a kidney cell, a neuronal cell, or another type of affected cell). ) from the subject a therapeutically effective amount of LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein), optionally with a pharmaceutically acceptable carrier. The LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein) implemented comprises a nucleotide sequence of interest useful for treating the disease. In particular, the TNA may comprise a desired exogenous DNA sequence operatively linked to control elements capable of directing transcription of the desired polypeptide, protein or oligonucleotide encoded by the exogenous DNA sequence when introduced into the subject. LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein) can be administered via any suitable route as described herein and known in the art. In one embodiment of any of the aspects or embodiments herein, the target cells are in a human subject. MA / t / zuzz / uoy / 01 Provided herein are methods of providing to a subject in need thereof a diagnostically or therapeutically effective amount of LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein), the The method comprises providing a cell, tissue or organ of a subject in need thereof, an amount of LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein); and for an effective time to allow expression of the transgene from the TNA LNPs, thereby providing the subject with a diagnostically or therapeutically effective amount of the protein, peptide, nucleic acid expressed by the TNA LNPs (e.g., lipid particles ( e.g., lipid nanoparticles) of the ceDNA vector as described herein). In one embodiment of any of the aspects or embodiments herein, the subject is a human being. Provided herein are methods for diagnosing, preventing, treating or ameliorating at least one or more symptoms of a disease, disorder, dysfunction, injury, abnormal condition or trauma in a subject. Generally, the method includes at least the step of administering to a subject in need thereof LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein), in an amount and during a time sufficient to diagnose, prevent, treat or improve one or more symptoms of the disease, disorder, dysfunction, injury, abnormal condition or trauma in the subject. In one embodiment of any of the aspects or embodiments herein, the subject is a human being. Provided herein are methods comprising the use of LNP with TNA as a tool to treat or reduce one or more symptoms of a disease or disease states. There are a number of inherited diseases in which defective genes are known, and they typically fall into two classes: deficiency states, usually enzymes, which are generally inherited recessively, and unbalanced states, which may involve regulatory or structural proteins. and which are usually, but not always, dominantly inherited. For deficiency state diseases, LNPs with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector as described herein) can be used to deliver transgenes to deliver a normal gene to affected tissues. for replacement treatment, as well as, in some embodiments of any of the aspects and modalities herein, to create animal models for the disease through antisense mutations. For imbalanced disease states, LNP with TNA (e.g., ceDNA vector lipid particles) can be used to create a disease in a model system, which could then be used to counteract the disease state. Therefore, LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) from the ceDNA vector) and the methods described herein allow the treatment of genetic diseases. As used herein, a disease is treated by partially or completely remedying the deficiency or imbalance that causes the disease or makes it more severe. In general, LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector) can be used to deliver any transgene according to the above description to treat, prevent or improve symptoms associated with any related disorder. with gene expression. Illustrative diseases include, but are not limited to: cystic fibrosis (and other lung diseases), hemophilia A, hemophilia B, thalassemia, anemia and other blood disorders, AIDS, Alzheimer's disease, Parkinson's disease, Huntington's chorea, amyotrophic lateral sclerosis , epilepsy and other neurological disorders, cancer, diabetes mellitus, muscular dystrophies (e.g. Duchenne, Becker), Hurler's disease, adenosine deaminase deficiency, metabolic diseases, retinal degenerative diseases (and other eye diseases), mitochondrial diseases ( e.g., Leber hereditary optic neuropathy (LHON), Leigh syndrome, and subacute sclerosing encephalopathy), myopathies (e.g., facioscapulohumeral myopathy (FSHD) and cardiomyopathies), diseases of solid viscera (e.g., brain, liver, kidney, heart ) and the like. In some embodiments of any of the aspects and embodiments herein, ceDNA vectors as described herein may be used advantageously in the treatment of individuals with metabolic disorders (e.g., ornithine transcarbamylase deficiency). In one embodiment of any of the aspects or modalities herein, the LNPs with TNA described herein can be used to treat, ameliorate and / or prevent a disease or disorder caused by mutation in a gene or gene product. Illustrative diseases or disorders that can be treated with TNA LNPs (e.g., ceDNA vector lipid particles (e.g., lipid nanoparticles) as described herein) include, but are not limited to, metabolic diseases or disorders (e.g. (e.g., Fabry disease, Gaucher disease, phenylketonuria (PKU), glycogen storage disease); diseases or disorders of the urea cycle (for example, ornithine transcarbamylase (OTO) deficiency); lysosomal storage diseases or disorders (e.g., metachromatic leukodystrophy (MLD), mucopolysaccharidosis type II (MPSII; Hunter syndrome)); liver diseases or disorders (for example, progressive familial intrahepatic cholestasis (PFIC); blood diseases or disorders (for example, hemophilia (A and B), thalassemia, and anemia); cancers and tumors, and genetic diseases or disorders (for example, cystic fibrosis). In one embodiment of any of the aspects or embodiments herein, LNPs with TNA (e.g., a lipid particle (e.g., lipid nanoparticles) of a ceDNA vector) can be used to deliver a heterologous nucleotide sequence in situations where for which it is convenient to regulate the level of transgene expression (for example, transgenes that encode hormones or growth factors). MA / IZ / ZUZZ / UOU101 In one embodiment of any of the aspects or embodiments herein, LNPs with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector) can be used to correct an abnormal level and / or function of a product. gene (for example, an absence or defect in a protein) results in the disease or disorder. LNPs with TNA (e.g., lipid particles (e.g., lipid nanoparticles) from the ceDNA vector) can produce a functional protein and / or modify protein levels to alleviate or reduce symptoms resulting from, or confer benefit to, a particular disease or disorder caused by the absence or defect in the protein. For example, treatment of OTO deficiency can be achieved by producing functional OTO enzyme; treatment of hemophilia A and B can be achieved by modifying the levels of Factor VIII, Factor IX and Factor X; Treatment of PKU can be achieved by modifying the levels of the enzyme phenylalanine hydroxylase; treatment of Fabry or Gaucher disease can be achieved by producing functional alpha galactosidase or beta glucocerebrosidase, respectively; treatment of MFD or MPSII can be achieved by producing functional arylsulfatase A or iduronate-2-sulfatase, respectively; treatment of cystic fibrosis can be achieved by producing a functional cystic fibrosis transmembrane conductance regulator; Treatment of glycogen storage disease can be achieved by restoring functional G6Pase enzyme function; and PFIC treatment can be achieved by producing functional ATP8B1, ABCB11, ABCB4, or TJP2 genes. In one embodiment of any of the aspects or embodiments herein, LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector) can be used to deliver an RNA-based therapeutic agent to a cell. in vitro or in vivo. Examples of RNA-based therapeutic agents include, but are not limited to, mRNA, antisense RNA and oligonucleotides, ribozymes, aptamers, RNA interference (RNAi), Dicer substrate dsRNA, small hairpin RNA (shRNA), asymmetric interference (iaRNA), microRNA (miRNA). For example, LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector) can be used to deliver an antisense nucleic acid to a cell in vitro or in vivo. For example, when the transgene is an RNAi molecule, expression of the antisense nucleic acid or RNAi in the target cell decreases the expression of a particular protein by the cell. Accordingly, transgenes that are RNAi molecules or antisense nucleic acids can be administered to decrease the expression of a particular protein in a subject in need. Antisense nucleic acids can further be administered to cells in vitro to regulate cell physiology, for example, to optimize cell or tissue culture systems. In one embodiment of any of the aspects or embodiments herein, LNP with TNA (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector) can be used to provide a DNA-based therapy to a cell in vitro or in vivo. The examples of MA / IZ / ¿U¿¿ / UO3101 DNA-based therapeutic options include, but are not limited to, minicircle DNA, minigene, viral DNA (e.g., AAV or lentivirus genome), or non-viral synthetic DNA vectors , closed-end linear duplex DNA (ceDNA / CELiD), plasmids, bacmids, doggybone™ DNA vectors, minimalist immunologically defined gene expression (MIDGE) vector, non-viral ministrand DNA vector (covalently closed linear DNA vector ) or minimal dumbbell-shaped DNA vector (dumbbell DNA). For example, in one embodiment of any of the aspects or embodiments herein, ceDNA vectors (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector) can be used to provide minicircles to a cell in vitro or in alive. For example, when the transgene is a minicircle DNA, expression of the minicircle DNA in the target cell decreases the expression of a particular protein by the cell. Consequently, transgenes that are minicircle DNA can be administered to decrease the expression of a particular protein in a subject in need. Minicircle DNAs can further be administered to cells in vitro to regulate cell physiology, for example, to optimize cell or tissue culture systems. In one embodiment of any of the aspects or embodiments herein, exemplary transgenes encoded by a TNA vector comprising an expression cassette include, but are not limited to; X, lysosomal enzymes (for example, hexosaminidase A, associated with Tay-Sachs disease, or sulfatase iduronate, associated with Hunter / MPS II syndrome), erythropoietin, angiostatin, endostatine, superoxide dysmutase, globin, leptin, catalase, catalase, catalase, catalase, catalase, catalase. tyrosine hydroxylase, as well as cytokines (for example, an interferon, β-interferon, interferon-y, interleukin-2, interleukin-4, interleukin 12, granulocyte-macrophage colony-stimulating factor, lymphotoxin and the like), growth factors of peptides and hormones (e.g., somatotropin, insulin, insulin-like growth factors 1 and 2, platelet-derived growth factor (PDGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), nerve growth factor (NGF), neurotrophic factor 3 and 4, brain-derived neurotrophic factor (BDNF), glia-derived growth factor (GDNF), transforming growth factor a and b and the like), receptors (e.g., nerve growth factor receptor tumor necrosis). In some illustrative embodiments, the transgene encodes a monoclonal antibody specific for one or more desired targets. In some illustrative embodiments, the ceDNA vector encodes more than one transgene. In some illustrative embodiments, the transgene encodes a fusion protein comprising two different polypeptides of interest. In some embodiments of any of the aspects and embodiments herein, the transgene encodes an antibody, including a full-length antibody or an antibody fragment, as defined herein. In some embodiments of any of the aspects and embodiments herein, the antibody is an antigen binding domain or an immunoglobulin variable domain sequence, as defined herein. Other illustrative transgenic sequences encode suicide gene products MA / IZ / ¿U¿¿ / UO3 / 01 (thymidine kinase, cytosine deaminase, diphtheria toxin, cytochrome P450, deoxycytidine kinase and tumor necrosis factor), proteins that confer resistance to a drug used in cancer treatment and products tumor suppressor genes. Administration In one embodiment of any of the aspects or embodiments herein, an LNP with TNA (e.g., a ceDNA vector lipid particle as described herein) can be administered to an organism for transduction of cells in vivo. In one embodiment of any of the aspects or embodiments herein, LNP with TNA (e.g., ceDNA vector lipid particles) can be administered to an organism for ex vivo cell transduction. Generally, administration is carried out by any of the routes normally used to introduce a molecule into maximum contact with blood or tissue cells. Suitable methods for administering such nucleic acids are available and well known to those skilled in the art, and, although more than one route can be used to administer a particular composition, a particular route can often provide a more immediate and effective reaction. what another way. Illustrative modes of administration of TNA LNPs (e.g., ceDNA vector lipid particles) include oral, rectal, transmucosal, intranasal, inhalation (e.g., via aerosol), buccal (e.g., sublingual), vaginal, intrathecal, infraocular, transdermal, intraendothelial, in utero (or ovum), parenteral (e.g., intravenous, subcutaneous, intradermal, intracranial, intramuscular [including administration to skeletal muscle, diaphragm and / or myocardium], intrapleural, intracerebral and intra-articular), topical (for example, both to the skin and mucosal surfaces, including respiratory tract surfaces and transdermal administration), intraphatic and the like, as well as direct injection into the tissue or organ (for example, to the liver, eye, skeletal muscle, myocardium, diaphragm or brain). Administration of the LNP with TNA as the ceDNA vector (e.g., ceDNA LNP) can be performed at any site in a subject, including, without limitation, a site selected from the group consisting of the brain, a skeletal muscle, a smooth muscle, the heart, the diaphragm, the epithelium of the respiratory tract, the liver, the kidney, the spleen, the pancreas, the skin and the eye. In one embodiment of any of the aspects or embodiments herein, administration of the ceDNA LNP may further be to a tumor (e.g., within or near a tumor or a lymph node). The most appropriate route in any case will depend on the nature and severity of the condition being treated, ameliorated and / or prevented and the nature of the particular ceDNA LNP being used. Additionally, ceDNA allows more than one transgene to be delivered in a single vector or multiple ceDNA vectors (e.g., a ceDNA cocktail). In one embodiment of any of the aspects or embodiments herein, administration of ceDNA LNP to skeletal muscle includes, but is not limited to, administration to skeletal muscle in the extremities (e.g., upper arm, lower part of the arm, upper leg and / or lower leg), back, neck, head (e.g. tongue), thorax, abdomen, pelvis / perineum and / or fingers. The ceDNA vectors (e.g., lipid particles (e.g., lipid nanoparticles) of the ceDNA vector) can be delivered to skeletal muscle by intravenous administration, intra-arterial administration, intraperitoneal administration, limb perfusion (optionally, isolated limb perfusion of a leg and / or arm; see, for example, Arruda et al., (2005) Blood 105: 3458-3464), and / or direct intramuscular injection. In particular embodiments, the ceDNA LNP is administered to an extremity (arm and / or leg) of a subject (e.g., a subject with muscular dystrophy such as DMD) by extremity perfusion, optionally isolated extremity perfusion (e.g., by intravenous or intra-articular administration). In one embodiment of any of the aspects or embodiments herein, the ceDNA LNP can be administered without employing hydrodynamic techniques. Administration of LPNs with TNA (e.g. ceDNA LNP) to the myocardium includes administration to the left atrium, right atrium, left ventricle, right ventricle and / or septum. TNA LNPs (e.g., ceDNA LPNs) can be delivered to the cardiac muscle by intravenous administration, intra-arterial administration, such as intra-aortic administration, direct cardiac injection (e.g., into the left atrium, right atrium, left ventricle, the right ventricle), and / or perfusion of the coronary artery. Administration to the diaphragm may be carried out by any suitable method, including intravenous administration, intra-arterial administration and / or intraperitoneal administration. Administration to smooth muscle can be carried out by any suitable method, including intravenous administration, Intraarterial administration and / or intraperitoneal administration. In one embodiment of any of the aspects or embodiments herein, administration may be to endothelial cells present in, near and / or on smooth muscle. In one embodiment of any of the aspects or embodiments herein, TNA LNPs (e.g., ceDNA LNPs) are administered to skeletal muscle, diaphragm, and / or myocardium (e.g., to treat, improve, and / or or prevent muscular dystrophy or heart disease (for example, PAD or congestive heart failure). LNPs with TNA (e.g., ceDNA LNPs) can be administered to the CNS (e.g., brain or eye). TNA LNPs (e.g. ceDNA LNPs) can enter the spinal cord, brainstem (medulla oblongata, pons), midbrain (hypothalamus, thalamus, epithelium, pituitary gland, substantia nigra, pineal gland), cerebellum, telencephalon (striatum, brain that includes the occipital, temporal, parietal and frontal lobes, cortex, basal ganglia, M A / t / zuzz / uoy l OI hippocampus and portaamygdala), limbic system, neocortex, striatum, brain and inferior colliculus. TNA LNPs (e.g. ceDNA LNPs) can be delivered to different regions of the eye, such as the retina, cornea and / or optic nerve. TNA LNPs (e.g. ceDNA LNPs) can be delivered into the cerebrospinal fluid (e.g. via lumbar puncture). LNPs with TNA (e.g., ceDNA vector lipid particles) can be additionally administered intravascularly to the CNS in situations where the blood-brain barrier has been disrupted (e.g., brain tumor or stroke). In one embodiment of any of the aspects or embodiments herein, TNA LNPs (e.g., ceDNA LNPs) can be administered to the desired regions of the CNS by any route known in the art, including, but not limited to, administration intrathecal, infraocular, intracerebral, intraventricular, intravenous (e.g., in the presence of a sugar, such as mannitol), intranasal, intraaural, infraocular (e.g., intravitreal, subretinal, anterior chamber), and periacular (e.g., region below Tenon's capsule), as well as intramuscular delivery with retrograde delivery to motor neurons. According to some embodiments of any of the aspects or embodiments herein, LNPs with TNA (e.g., ceDNA LNPs) are administered in a liquid formulation by direct injection (e.g., stereotactic injection) into the desired region or compartment. in the CNS. According to other embodiments, LNPs with TNA (e.g., ceDNA LNPs) can be provided by topical application to the desired region or by intranasal administration of an aerosol formulation. Administration to the eye can be by topical application of liquid drops. As a further alternative, the ceDNA vector can be administered as a solid slow release formulation (see, for example, US Patent 7,201,898, which is incorporated herein by reference in its entirety). In one embodiment of any of the aspects or embodiments herein, LNPs with TNA (e.g., ceDNA LNPs) can be used for retrograde transport to treat, ameliorate and / or prevent diseases and disorders involving motor neurons (e.g. , amyotrophic lateral sclerosis (ALS); spinal muscular atrophy (SMA), etc.). For example, LNPs with TNA (e.g. ceDNA LNPs) can be delivered to muscle tissue from which it can migrate to neurons. In one embodiment of any of the aspects or embodiments herein, repeated administrations of the therapeutic product may be performed until the appropriate level of expression is achieved. Therefore, in one embodiment of any of the aspects or embodiments herein, a therapeutic nucleic acid can be administered and re-dosed multiple times. For example, the therapeutic nucleic acid may be administered on day 0. After the initial treatment on day 0, a second dosing (redosing) may be performed in about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks. , approximately 6 weeks, approximately 7 weeks, approximately 8 weeks, or approximately 3 months, approximately 4 months, approximately 5 months, approximately 6 months, approximately 7 months, approximately 8 months, approximately 9 months, ; approximately 10 months, approximately 11 months, or approximately 1 year, approximately 2 years, approximately 3 years, approximately 4 years, approximately 5 years, approximately 6 years, approximately 7 years, approximately 8 years, approximately 9 years, approximately 10 years, approximately 11 years, approximately 12 years, approximately 13 years, approximately 14 years, approximately 15 years, approximately 16 years, approximately 17 years, approximately 18 years, approximately 19 years, approximately 20 years, approximately 21 years, approximately 22 years, approximately 23 years , approximately 24 years, approximately 25 years, approximately 26 years, approximately 27 years, approximately 28 years, approximately 29 years, approximately 30 years, approximately 31 years, approximately 32 years, approximately 33 years, approximately 34 years, approximately 35 years, approximately 36 years, approximately 37 years, approximately 38 years, approximately 39 years, approximately 40 years, approximately 41 years, approximately 42 years, approximately 43 years, approximately 44 years, approximately 45 years, approximately 46 years, approximately 47 years, approximately 48 years , approximately 49 years or approximately 50 years after initial treatment with the acid MA / IZ / ¿U¿¿ / UO3 / 01 therapeutic nucleic. In an embodiment of any of the aspects or embodiments herein, one or more additional compounds may also be included. Those compounds may be administered separately or additional compounds may be included in the lipid particles (e.g., lipid nanoparticles) of the invention. In other words, lipid particles (e.g., lipid nanoparticles) may contain other compounds in addition to TNA or at least a second TNA, different from the first. Without limitation, other additional compounds may be selected from the group consisting of organic or inorganic molecules, small or large, monosaccharides, disaccharides, trisaccharides, oligosaccharides, polysaccharides, peptides, proteins, peptide analogs and derivatives thereof, peptidomimetics, nucleic acids, analogs of nucleic acids and derivatives, an extract made from biological materials, or any combination of these. In one embodiment of any of the aspects or embodiments herein, the one or more additional compounds may be a therapeutic agent. The therapeutic agent may be selected from any class suitable for the therapeutic objective. Accordingly, the therapeutic agent can be selected from any class suitable for the therapeutic objective. The therapeutic agent can be selected according to the objective of the treatment and the desired biological action. For example, in one embodiment of any of the aspects or embodiments herein, if the TNA within the LNP is useful for treating cancer, the additional compound may be an antineoplastic agent (e.g., a chemotherapeutic agent, a targeted treatment against cancer (including, but not limited to, a small molecule, an antibody or an antibody-drug conjugate). In one embodiment of any of the aspects or embodiments herein, if the LNP containing the TNA is useful to treat an infection, the additional compound may be an antimicrobial agent (for example, an antibiotic or antiviral compound). In one embodiment of any of the aspects or embodiments herein, if the LNP containing the TNA is useful to treat an disease or an immune disorder, the additional compound may be a compound that modulates an immune response (for example, an immunosuppressant, an immunostimulatory compound or a compound that modulates one or more specific immune pathways). In one embodiment of any of the aspects or embodiments herein, different cocktails of different lipid particles containing different compounds, such as a TNA encoding a different protein or a different compound, such as a therapeutic agent, can be used in the compositions. and methods of the invention. In one embodiment of any of the aspects or embodiments herein, the additional compound is an immunomodulatory agent. For example, the additional compound is an immunosuppressant. In some embodiments of any of the aspects or embodiments herein, the additional compound is immunostimulatory. EXAMPLES The following examples are provided by way of illustration, not limitation. One skilled in the art will appreciate that ionizable lipids can be designed and synthesized using general synthesis methods described below. General Summary The ionizable lipids of Formula I were designed and synthesized using similar synthesis methods depicted in Scheme 1 below. Scheme 1 MA. t / zuzz / uoy 101 ' OH .—üMs ' ' 3 2 1 cm Stage 2N-, / c IAO Stages hcA-T to you >-,J|| 'T and OH ' or} \ C \—q M and --- ·-?.. and -«· ' o o R'-OH T c υ _ U iiií; 11 o··:· b -* -C. _bjj- r- Stage 5 o EDO D'diP Stage 3 and Pyridine hf mi Stage 4 Example 1: Synthesis of 1-(heptadecan-9-yl)9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxyjphenyl)acetoxy)) nonanedioate )et¡Ijpiperidin-1 -yl)ethyl)d¡sultanoíI)etiI)piperidin-4-yl)ethoxy)-2-oxoethyl)pheniI) (Lipid 1) Synthesis of an ionizable and cleavable head group ((disulfanedi¡lb¡s(ethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl)bis(2)acetate -(4-hydroxyphenyl)) (7) Stage 1 MsCI NEt3—OMs Yes I s ^OMs Synthesis of disulfanediylbis(ethane-2,1-diyl) dimethanesulfonate (2). Commercially available 2,2'-disulfanediylbis(ethan-1-ol) (1) (15 g, 97.2 mmol) was dissolved in acetonitrile (143 mL) followed by the addition of NEts (33.3 g, 328 mmol). MsCl (34.5 g, 300 mmol) was added dropwise to the reaction mixture at 0 °C. The resulting reaction mixture was stirred at room temperature for 3 h. Ethanol (EtOH) (39 mL) was added to the reaction mixture to ΜΛ / t / ZUZZ / UOU / 01 quenched the reaction and insoluble materials were removed by filtration. The filtrate was partitioned between dichloromethane (DCM) (150 mL) and 10% sodium bicarbonate / water (150 mL). The organic layer was washed with 100 mL of water four times, dried over (MgSCU), and evaporated to give 2 as a brown oil (25 g, 81%), which solidified on standing.1H-NMR (300 MHz, d0 (m, 10H). Stage 2 chloroform): δ 4.43-4.48 (t, 4H), 3.00-3.1 —OMs Yes I s ^OMs Synthesis of 2,2'-((disulfanediylb¡s(ethane-2,1-di¡l))b¡s(piper¡dine-1,4-diyl))bis(ethane-1-ol) ( 4). To a solution of 2 (12 g, 38.7 mmol) in acetonitrile (310 mL) was added potassium carbonate (K2CO3) (13.4 g, 96.6 mmol) followed by 2-(piperidin-4-yl)ethane-1-ol ( 3) (20 g, 155 mmol). The resulting mixture was stirred at room temperature overnight before removing insoluble material by filtration. The filtrate was evaporated to dryness to give the crude product, which was dissolved in DCM (100 mL), washed with water twice (50 mL), dried over MgSCU and evaporated to give 4 as a yellow oil (11.8 g, 79 %).1H-NMR (300 MHz, d-chloroform): δ 3.63-3.68 (t, 4H), 2.78-2.90 (m, 8H), 2.62-2.65 (t, 4H), 1.94-2.02 (t, 4H), 1.70 (s, 2H), 1.65-1.70 (d, 4H), 1.27-1.48 (t, 4H), 1.40-1.50 (m, 2H), 1.23-1.27 (m, 4H). Stage 3 O O HO-^—OH ----HO^—^jy~OTBS 5a5 Synthesis of 2-(4-((tert-butyldimethylsilyl)oxy)phenyl)acetic acid (5). To a stirred solution of 4hydroxyphenylacetic acid (5a) (10 g, 65 mmol) in dimethylformamide (DMF) (40 mL) at 0 °C was added NEts (10 g, 100 mmol) followed by ether-butyldimethylsilyl chloride (TBSCI ) (15 g, 100 mmol). The resulting reaction mixture was stirred at room temperature overnight, then treated with water (200 mL) and DCM (150 mL). The organic phase was separated. The aqueous phase was extracted with DCM (100 mL). The combined organic phase was washed with saturated sodium bicarbonate solution, brine and dried over sodium sulfate (Na2SC>4). The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-10% methanol (MeOH) in DCM as eluent. Fractions containing the desired compound were combined and evaporated to provide 5 (4.8 g, 27%) and the byproduct di-tert-butyldimethylsilyl ether (di-TBS) (10.5 g, 42%).1H-NMR of 5 (300 MHz, d-chloroform): δ 7.12 (d, 2H), 6.78 (d, 2H), 3.56 (s, 2H), 0.97 (s, 9H), 0.18 (s, 6H). MA. t / zuzz / uoy lOI Synthesis of ((disulfanediylbis(ethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl) bis(2(4-((tert-butyldimethylsilyl))acetate oxy)phenyl)) (6). To a stirred solution of the disulfide 4 produced in step 2 (1.92 g, 5 mmol) and phenylacetic acid 5 (3.4 g, 12.8 mmol) in DCM (100 mL) was added 4-dimethylaminopyridine ( DMAP) (1.5 g, 12.5 mmol) followed by 1-ethyl-3-(3-dimethylaminopropylcarbodiimide (EDCI) (2.4 g, 12.5 mmol). The resulting mixture was stirred at room temperature overnight, then washed with a saturated solution of sodium bicarbonate (200 mL), brine (150 mL) and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. Fractions containing the desired compound were evaporated to give 6 (4.1 g, 92%).1H-NMR of 6 (300 MHz, d-chloroform): δ 7.12 (d, 4H), 6.75 (d, 4H), 4.1 (t, 4H), 3.5 (s, 4H), 2.82 (m, 8H), 2.62 (m, 4H), 1.93 (t, 4H), 1.61 -1.45 (m, 8H), 1.26 (m , 6H), 0.97 (s, 18H), 0.17 (s, 4H). Stage 4 ~lj / Ή, Ϊ > \ 1 >: {—' j —( ΗΓ6: í. —'' — Synthesis of ((disulfanediylbis(ethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl)bis(2-(4hydroxyphenyl)) acetate (7 ). To a stirred solution of disulfide 6 (3.1 g, 3.6 mmol) in tetrahydrofuran (THF) (40 mL) was added pyridine hydrogen fluoride (1 mL, 3.8 mmol) at 0 °C. The resulting mixture was stirred at 0 °C for 2 h, then at room temperature for another 2 h. The reaction mixture was treated with saturated sodium bicarbonate solution (200 mL) and extracted with ethyl acetate (2 x 150 mL). The combined organic phase was washed with brine (100 mL), dried over NazSCU and concentrated. The residue was purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent which gave the desired product 7 (1.92 g, 82%).1H-NMR (300 MHz, d chloroform ): δ 7.13 (d, 4H), 6.70 (d, 4H), 4.1 (t, 4H), 3.5 (s, 4H), 2.89 (m, 8H), 2.70 (m, 4H), 1.95 (t, 4H ), 1.48 (m, 8H), 1.17 (m, 6H). Synthesis of 9-(heptadecan-9-yloxy)-9-oxononano¡co acid (10) Synthesis of 9-(heptadecan-9-yloxy)-9-oxononano¡co acid (10). To a stirred solution of nonanedioic acid (8) (7.34 g, 39 mmol) and heptadecan-9-ol (8b) (5 g, 19 mmol) in dichloromethane (1000 mL) was added DMAP (2.37 g, 19 mmol) followed by by EDCI (3 g, 19 mmol). The resulting mixture was stirred at room temperature overnight, then washed with 250 mL of 1 N HCl and 250 mL of water. The organic layer was dried over MgSCU, evaporated to dryness, and purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. Fractions containing the desired compound were pooled and evaporated to give 10 (6.2 g, 75%) as a white solid.1H-NMR (300 MHz, d-chloroform): δ 4.80-4.90 (m, 1H), 2.25- 2.34 (m, 4H), 1.55-1.70 (m, 4H), 1.40-1.50 (m, 4H), 1.20-1.40 (m, 30H), 0.84-0.90 (t, 3H). Synthesis of 1-(heptadecan-9-yl)9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4hydroxyphenyl)acetoxy)ethyl)) nonanedioate piperidin-1 -yl)ethyl)disulfanoyl)ethyl)p¡peridin-4-yl)ethoxy)-2-oxoethyl)phenyl) Synthesis of del-(heptadecan-9-yl)9-(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-hydroxyphenyl)acetoxy)ethyl) nonanedioate) ) piperidin-1 -¡l)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (11). To a stirred solution of the disulfide 7 produced in Step 4 (580 mg, 0.9 mmol) and acid 10 (422 mg, 0.99 mmol) in DMF (20 mL) was added DMAP (165 mg, 1.35 mmol) followed by of EDCI (258 mg, 1.35 mmol). The resulting mixture was stirred at room temperature overnight and then a saturated sodium bicarbonate solution (50 mL) was added. The reaction mixture was extracted with dichloromethane (2 x 50 mL). The combined organic phase was washed with brine (30 mL), dried over NasSCU, and concentrated. The residue was purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent to give the desired product 11 (427 mg, 45%).1H-NMR (300 MHz, d-chloroform ): δ 7.27 (d, 2H), 7.11 (d, 2H), 7.03 (d, 2H), 6.69 (d, 2H), 4.85 (m, 1H), 4.1 (m, 4H), 3.56 (s, 2H ), 3.48 (s, 2H), 2.92 (d, 2H), 2.85-2.69 (m, 12H), 2.71 (t, 2H), 2.28 (t, 2H), 1.95 (t, 2H), 1.52-1.01 ( m, 53H), 0.85 (m, 6H). Lipid 1 Synthesis / \} \ .1. ,.Í X ,L.i. ( '· ' '· 3 ’* <·' Γ. '·-’ '' ''·' 'Γ.ι—k·. I—, ,·- --- / 'X- ÚH V12 / - _ i X ,¡j¿ ........... . / . ··>c· ··.-.·· ··...·· ··...M w --t' Lipid 1 Synthesis of 1-(heptadecan-9-yl)9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxy))phenyl)) nonanedioate acetoxy))ethyl)piperidín-1 -¡l)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (Lipid 1). To a stirred solution of disulfide 11 (151 mg, 0.14 mmol) and oleic acid 12 (61 mg, 0.22 mmol) in dichloromethane (10 mL) was added DMAP (28 mg, 0.22 mmol) followed by EDCI (42 mg, 0.22 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), and dried over NasSCL. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. Fractions containing the desired compound were evaporated to give Lipid 1 (126 mg, 68%). 1H-NMR of Lipid 1 (300 MHz, d-chloroform): δ 7.25 (d, 4H), 7.01 (d, 4H ), 5.34 (m, 2H), 4.86 (m, 1H), 4.11 (t, 4H), 3.58 (s, 4H), 2.91-2.70 (m, 8H), 2.62 (m, 4H), 2.53 (t, 4H), 2.28 (t, 2H), 2.05-1.87 (m, 8H), 1.781.46 (m, 22H), 1.48-1.23 (m, 54H), 0.86 (t, 9H). MS [M+H]+1318. Example 2: Synthesis of 1-(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-())) nonanedioate (9(nonyloxy)-9-oxononanoyl)oxy)phenyl)acetoxy)ethyl)piperid¡n-1-yl)ethyl)disulfanoyl)ethyl)p¡perídin-4-yl)ethoxy)-2oxoethyljphenyl) ( lipid 3) Synthesis of 9-(nonyloxy)-9-oxononano¡co acid (9) MA / t / zuzz / uoy101 Synthesis of 9-(nonyloxy)-9-oxononano¡co acid (9). To a stirred solution of nonanedioic acid (8) (13.2 g, 0.1 mol) and nonan-1-ol (8a) (7.2 g, 0.05 mol) in DCM (1000 mL) was added DMAP (6.1 g, 0.05 mol) followed by by EDCI (7.7 g, 0.05 mol). The resulting mixture was stirred at room temperature overnight, then washed with 1 N hydrochloric acid (HCl) solution (500 mL) and water (500 mL). The organic layer was dried over MgSCu, evaporated to dryness and purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. Fractions containing the desired compound were pooled and evaporated to give 9 (12.6 g, 81%) as a white solid.1H-NMR (300 MHz, d-chloroform): δ 4.03-4.07 (t, 2H), 2.28- 2.34 (m, 4H), 1.58-1.63 (m, 6H), 1,261.32 (m, 18H), 0.85-0.87 (t, 3H). Lipid 3 Synthesis Lipid 3 Synthesis of 1-(heptadecan-9-yl)9-(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-((9-))) nonanedioate (nonyloxy¡)-9oxononanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (Lipid 3). To a stirred solution of disulfide 11 (step synthesis described in Example 1) (150 mg, 0.14 mmol) and acid 9 (62 mg, 0.22 mmol) in dichloromethane (10 mL) was added DMAP (28 mg, 0.22 mmol ) followed by EDCI (42 mg, 0.22 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), and dried over NasSCU. The solvent was removed under reduced pressure and the residue was purified by Silica gel column chromatography using 0-10% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 3 (114 mg, 60%). 4.86 (m, 1H), 4.11 (t, 4H), 4.04 (t, 2H), 3.58 (s, 4H), 2.93-2.77 (m, 8H), 2.63 (m, 4H), 2.53 (t, 4H) , 2.28 (m, 4H), 1.95 (t, 4H), 1.85-1.47 (m, 24H), 1.45-1.16 (m, 54H), 0.86 (t, 9H). MS [M+H]+1350. Example 3: Synthesis of 1-(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-())) nonanodloate (5 -(nonyloxy)5-oxopentanoyl)oxy)phenyl)acetoxy¡)ethyl) piperidin-1 -i l)ethyl)disulfanoyl)ethyl) piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (Lipid 2) ü.... ......... .. k. γ ··_..Ϋ . / _r = 'r ' Γ ' r ’ Lipid 2 To a stirred solution of disulfide 11 (step synthesis described in Example 1) (150 mg, 0.14 mmol) and acid 9a (see the synthesis described in Example 1 for acid 9, where nonanedioic acid (8) was replaced with commercially available glutaric acid as starting material to react with nonan-1-ol (8a) to produce 9a) (57 mg, 0.22 mmol) in DCM (10 mL) DMAP (28 mg, 0.22 mmol) was added followed by EDCI (42 mg, 0.22 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), and dried over NaaSCU. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 2 (151 mg, 81%). 1H-NMR of Lipid 2 (300 MHz, d-chloroform): δ 7.26 (d, 4H), 7.01 (d, 4H ), 4.86 (m, 1H), 4.10-4.02 (t, 6H), 3.57 (s, 4H), 3.01 (d, 4H), 2.83-2.72 (m, 4H), 2.34-2.21 (m, 14H), 2.15-1.91 (m, 6H), 1.74-1.41 (m, 12H), 1.39-1.16 (m, 52H), 0.86 (t, 9H). MS [M+H]+1293. Example 4: Synthesis of 1-(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-())) nonanodloate (5(nonyloxy)-5-oxopentanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1-yl)ethyl)disulfanoyl)ethyl)piper¡din-4-yl)ethoxy¡)-2-oxoethyl) phenyl) (lipid 4)vi N '>----'' Π To a stirred solution of disulfide 7 (step synthesis described in Example 1) (150 mg, 0.23 mmol) and compound 9 (synthesis described in Example 2) (146 mg, 0.46 mmol) in a mixture of dichloromethane (5 mL ) and DMF (3 mL) DMAP (70 mg, 0.57 mmol) was added followed by EDCI (109 mg, 0.57 mmol) at 0 °C. The resulting mixture was stirred at 0°C for 15 minutes, then at room temperature overnight. DCM (20 mL) was added and the reaction mixture was washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), dried over NazSCU. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 4 (180 mg, 63%). 4.11 (t, 4H), 4.04 (t, 4H), 3.58 (s, 4H), 2.93-2.67 (m, 8H), 2.63-2.55 (m, 4H), 2.53 (t, 4H), 2.29 (t, 4H), 1.94 (t, 4H), 1.85-1.47 (m, 20H), 1.45-1.16 (m, 42), 0.87 (t, 6H). MS [M+H]+1237. Example 5: Synthesis of 9,9'-di(heptadecan-9-yl) di(nonanedioate) from 0'1,01((((((disulfanedi¡lb¡s(ethane-2,1-di¡lo)) )bis(piperidine-1,4-dyl))bis(ethane-2,1-diyl))bis(oxy))bis(2oxoethane-2,1-diyl))bis(4,1-phenylene)) (Lipid 5) U .o, -,Á · · - J Lipid 5 ΜΛ. t / zuzz / uoy lOI To a stirred solution of disulfide 7 (step synthesis as described in Example 1) (580 mg, 0.9 mmol) and acid 10 (synthesis described in Example 1) (422 mg, 0.99 mmol) in DMF (20 mL) was added DMAP (164 mg, 1.35 mmol) followed by EDCI (257 mg, 1.35 mmol) at 0 °C. The resulting mixture was stirred at 0°C for 15 minutes, then at room temperature overnight. DCM (60 mL) was added and the reaction mixture was washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), dried over NazSCU. The solvent was removed under reduced pressure and the residue was purified by chromatography. on silica gel column by using 010% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 5 (280 mg, 38%). 1H-NMR of Lipid 5 (300 MHz, d-chloroform): δ 7.26 (d, 4H), 7.02 (d, 4H ), 4.85 (m, 2H), 4.11 (t, 4H), 3.58 (s, 4H), 2.86-2.77 (m, 8H), 2.63 (m, 4H), 2.53 (t, 4H), 2.27 (t, 4H), 1.92 (t, 4H), 1.75-1.47 (m, 26H), 1.45-1.16 (m, 64H), 0.86 (t, 12H). MS [M+H]+1462. Example 6: Synthesis of 9-(undecan-3-yl)nonanedioate from 1-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxy) (lipid 6) Synthesis of 9-oxo-9-(undecan-3-yloxy)nonanoic acid (9b) Synthesis of 9-oxo-9-(undecan-3-yloxy)nonanoic acid (9b). To a stirred solution of nonanedioic acid (8) (10.9 g, 0.058 mol) and undecan-3-ol (8b) (5 g, 0.029 mol) in DCM (500 mL) was added DMAP (3.5 g, 0.03 mol) followed by by EDCI (4.5 g, 0.03 mol). The resulting mixture was stirred at room temperature overnight, then washed with a 1 N HCl solution (500 mL) and water (500 mL). The organic layer was dried over MgSCu, evaporated to dryness and purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. Fractions containing the desired compound were pooled and evaporated to give 9b (6.5 g, 66%) as a white solid.1H-NMR (300 MHz, d-chloroform): δ 4.79-4.83 (t, 1 H), 2.28 -2.34 (m, 4 H), 1.25-1.33 (m, 8 H), 1.26-1.32 (m, 18 H), 0.85-0.87 (t, 6 H). Synthesis of 4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-hydroxyphenyl)acetoxy)ethyl)piperidin-1yl)ethyl)disulfanoyl)ethyl oleate piperidin-4-¡l)ethoxy¡)-2-oxoethyl)pheníl (13). / '·,___;·' Oleic acid —. ·::\¡ ,-ri , n -r i .—,u—, H ... ...... - '-Yo. ; '----JH .· '----' -J- --- --- --' ---' -- -- ---Á * i_ L > -· X t,· > Λ-J713 Synthesis of 4-(2-(2-(1-(2-((2-(4-(2-(2-(4-hydroxyphenyl)acetoxy)ethyl)piperidin-1yl)ethyl)disulfanoyl)ethyl oleate piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl (13). To a stirred solution of disulfide 7 (step synthesis described in Example 1) (2.0 g, 3 mmol) and oleic acid (or acid 12 described in Example 1) (0.79 g, 2.8 mmol) in DCM (200 mL) DMAP (340 mg, 2.8 mmol) was added followed by EDCI (440 mg, 2.8 mmol). The resulting mixture was stirred at room temperature overnight and then a saturated sodium bicarbonate solution (20 mL) was added. The reaction mixture was extracted with dichloromethane (2 χ 50 mL). The combined organic phase was washed with brine (30 mL), dried over NazSCU and concentrated. The residue was purified by silica gel column chromatography using 0-5% methanol in dichloromethane as eluent to provide 13 (1.6 g, 57%) as a white solid. The product was used directly in the next step without additional characterization. Lipid 6 Synthesis Lipid 6 Synthesis of 1-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxy)phenyl))) 9-(undecan-3-yl)nonanedioate acetoxy)ethyl)piperidin-1 -II)ethyl)dísulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (Lipid 6). To a stirred solution of disulfide 13 (250 mg, 0.27 mmol) and acid 9b (113 mg, 0.33 mmol) in DCM (20 mL) was added DMAP (40 mg, 0.33 mmol) followed by EDCI (51 mg, 0.33 mmol) . The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-5% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 6 (120 mg, 36%). 1H-NMR (300 MHz, d-chloroform): δ 7.31 (d, 4 H), 7.05 (d, 4 H) , 5.36-5.40 (m, 2 H), 4.86 (m, 1 H), 4.11 (t, 4 H), 3.62 (t, 4 H), 2.77-2.90 (m, 8 H), 2,552.71 (m , 8 H), 2.30-2.34 (m, 2 H), 1.96-2.05 (m, 8 H), 1.77 (m, 4 H), 1.58-1.67 (m, 18 H), 1.30-1.58 (m, 40 H), 0.89 (t, 9 H). Example 7: Synthesis of 9-(tridecan-5-yl)nonanedioate from 1-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxy) )phen¡l)acetoxy)ethyl)p¡peridin-1) -¡l)ethyl)disulfan¡l)ethyl)piperidín-4-yl)ethoxy)-2-oxoethyl)phenyl) (Lipid 7) Synthesis of tridecanol-5-ol (8c) 8c Synthesis of tridecan-5-ol (8c). To a solution of aldehyde 8c-1 (7.1 g, 0.05 mol) in 100 mL of anhydrous THF was added dropwise at −78 °C a solution of 2 M butyl lithium (BuLi) (27 mL) in THF. The resulting mixture was stirred at -78°C for 2 hours and then at room temperature for 2 hours. The reaction was quenched by addition of water and partitioned between 1 N HCl and ether. The organic layer was collected, dried over MgSO4, and evaporated to give crude 8c (10 g, 100%) as a yellow oil, which was used directly for the next step without further purification. Synthesis of 9-oxo-9-(tridecan-5-yloxy)nonano¡ic acid (9c) MA / IZ / ¿U¿¿ / UO31 OI O O EDCI q o HEE HEE 1 DMAPrU - . . U+HOHOO 8 8c 9c Synthesis of 9-oxo-9-(tridecan-5-ylox¡)nonano¡co acid (9c). To a stirred solution of nonanedioic acid (8) (9.4 g, 0.05 mol) and 8c (5 g, 0.025 mol) in DCM (500 mL) was added DMAP (3.05 g, 0.025 mol) followed by EDCI (3.88 g, 0.025 mole). The resulting mixture was stirred at room temperature overnight, then washed with a 1 N HCl solution (500 mL) and water (500 mL). The organic layer was dried over MgSO4, evaporated to dryness and purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. The fractions containing the desired compound were pooled and evaporated to give 9c (2.5 g, 27%) as a white solid.1H-NMR (300 MHz, d-chloroform): δ 4.84-4.87 (t, 1 H), 2.282 .34 (m, 4 H), 1.58-1.63 (m, 7 H), 1.26-1.32 (m, 23 H), 0.85-0.87 (t, 6 H). Lipid 7 Synthesis Lipid 7 Synthesis of 1-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4-(oleo¡lox¡)phen¡l)acetoxy)etoxy)et) nonanedioate ¡l)p¡peñdin-1-¡l) 9-(tridecan-5-yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (Lipid 7). To a stirred solution of disulfide 13 (synthesis described in Example 6) (250 mg, 0.27 mmol) and acid 9c (116 mg, 0.33 mmol) in DCM (20 mL) was added DMAP (40 mg, 0.33 mmol) followed by EDCI (51 mg, 0.33 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-5% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 7 (160 mg, 40%).1H-NMR (300 MHz, dchloroform): δ 7.29 (d, 4 H), 7.04 (d, 4 H), 5.29 -5.34 (m, 2 H), 4.86 (m, 1 H), 4.11 (t, 4 H), 3.58 (t, 4 H), 2.77-2.90 (m, 8 H), 2.51-2.79 (m, 8 H), 2.28 (m, 2 H), 1.94-2.05 (m, 8 H), 1.70-1.80 (m, 4 H), 1.49-1.67 (m, 18 H), 1.10-1.40 (m, 46 H) , 0.88 (t, 9 H). Example 8: Synthesis of 9-(pentadecan-7-yl)nonanedioate from 1-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxy) )phenyl)acetoxy¡)ethyl)p¡peridin-1) -¡l)ethyl)disulfan¡l)ethyl)p¡pendin-4-yl)ethoxy¡)-2-oxoethyl)phenyl) (lipid 8) Synthesis of pentadecan-7-ol (8d) 8d iviA / t / zuzz / uoy i heard Synthesis of pentadecan-7-ol (8d). To a solution of aldehyde 8d-1 (7.1 g, 0.05 mol) in 100 mL of anhydrous THF was added a solution of 2 M hexylmagnesium bromide in THF (27 mL) at −78 °C. The resulting mixture was stirred at -78°C for 2 hours and then at room temperature overnight. The reaction was quenched by addition of water and partitioned between 1 N HCl and ether. The organic layer was collected, dried over MgSCh, and evaporated to give crude 8d (11 g, 100%) as a white solid, which was used directly for the next step without further purification. Synthesis of 9-oxo-9-(pentadecan-7-yloxy)nonanoic acid (9d) EDCI DMAP 9d Synthesis of 9-oxo-9-(pentadecan-7-yloxy)nonanoic acid (9d). To a stirred solution of nonanedioic acid (8) (9.4 g, 0.05 mol) and pentadecane-7-ol (8d) (5.7 g, 0.025 mol) in DCM (1000 mL) was added DMAP (3.05 g, 0.025 mol) followed by by EDCI (3.88 g, 0.025 mol). The resulting mixture was stirred at room temperature overnight, then washed with a 1 N HCl solution (500 mL) and water (500 mL). The organic layer was dried over MgSCU, evaporated to dryness, and purified by silica gel column chromatography using 0-10% MeOH in DCM as eluent. Fractions containing the desired compound were pooled and evaporated to give 9d (6.2 g, 62%) as a white solid.1H-NMR (300 MHz, d-chloroform): δ 4.86 (t, 1 H), 2.28-2.34 (m, 4 H), 1.58-1.63 (m, 8 H), 1.26-1.32 (m, 27 H), 0.85-0.87 (t, 6 H). Lipid 8 Synthesis 100 MA / t / zuzz / uoy l OI Lipid 8 Synthesis of 1-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(oleoyloxy)phenyl))) 9-(pentadecan-7-yl)nonanedioate acetoxy)ethyl)piperidin-1-yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (Lipid 8). To a stirred solution of disulfide 13 (synthesis described in Example 6) (250 mg, 0.27 mmol) and acid 9d (120 mg, 0.33 mmol) in DCM (20 mL) was added DMAP (40 mg, 0.33 mmol) followed by EDCI (51 mg, 0.33 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-5% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 8 (170 mg, 40%).1H-NMR (300 MHz, d-chloroform): δ 7.29 (d, 4 H), 7.04 (d, 4 H) , 5.29-5.34 (m, 2 H), 4.86 (m, 1 H), 4.11 (t, 4 H), 3.58 (t, 4 H), 2.80-2.93 (m, 8 H), 2.51-2.68 (m , 8 H), 2.28 (m, 2 H), 1.97-2.05 (m, 8 H), 1.70-1.80 (m, 4 H), 1.50-1.70 (m, 18 H), 1.10-1.40 (m, 58 H), 0.87 (t, 9 H). Example 9: Synthesis of 1-nonyl 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-(((9)) nonanedioate -oxo -9-(undecan3-yloxy)nonanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -¡l)ethyl)disulfanoyl)ethyl)piperidin-4-¡l)ethoxy¡)ethyl)phenyl) ( Lipid 9) Synthesis of 9-nonyl nonanedioate from 1 -(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-hydroxy¡phenyl)acetoxy)etoxy)et l)piperidin-1yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2-oxoethyl)phenyl) (14). Synthesis of 1 -(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-hydroxyphen¡l)acetoxy¡)ethyl)piperidin) 9-nonyl nonanedioate) -yl)ethyl)disulfanoyl)ethyl)piperidin-4-íl)ethoxy)-2-oxoethyl)pheníl) (14). To a stirred solution of disulfide 7 (step synthesis described in Example 1) (3.1 g, 4.8 mmol) and 9-(nonyloxy)-9-oxononanoic acid (9) (synthesis described in Example 1) (1.51 g, 4.8 mmol) in dichloromethane (200 mL) was added 101 DMAP (587 mg, 4.8 mmol) followed by EDCI (746 mg, 4.8 mmol). The resulting mixture was stirred at room temperature overnight, then saturated sodium bicarbonate solution (50 mL) was added. The reaction mixture was extracted with dichloromethane (2 χ 50 mL). The combined organic phase was washed with brine (30 mL), dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography using 0-5% MeOH in DCM as eluent to give the desired product 14 (2.47 g, 55%). The product was used directly in the next step without additional characterization. Lipid 9 Synthesis Ί 14 / Ό v. TO. Aj® A 1 Lipid 9 Synthesis of 1-nonyl nonanedioate 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-((9-oxo- 9- (undecan-3yloxy)nonanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-íl)ethoxy)ethyl)phenyl) (Lipid 9) . To a stirred solution of disulfide 14 (250 mg, 0.26 mmol) and acid 9b (synthesis described in Example 6) (110 mg, 0.32 mmol) in dichloromethane (20 mL) DMAP (46 mg, 0.37 mmol) was added. followed by EDCI (50 mg, 0.32 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL), and dried over Na2SC>4. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-5% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 9 (230 mg, 68%). 1H-NMR (300 MHz, d-chloroform): δ 7.28 (d, 4 H), 7.04 (d, 4 H) , 4.86 (m, 1 H), 4.06-4.12 (t, 4 H), 4.04 (t, 2 H), 3.59 (s, 4 H), 2.60-2.90 (m, 8 H), 2.27-2.60 (m , 10 H), 1.97 (t, 3 H), 1.52-1.80 (m, 18 H), 1.10-1.40 (m, 40 H), 0.88 (t, 9 H). Example 10: Synthesis of 1-nonyl 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-(((9)) nonanedioate -oxo -9(tridecan-5-¡lox¡)nonanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfan¡l)ethyl)p¡peridin-4¡l)ethoxy)ethyl )phenyl) (Lipid 10) 102 M A / t / zuzz / uoy l OI Lipid 10 Synthesis of 1-nonyl nonanedioate 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-((9-oxo- 9- (tridecan-5yloxy)nonanoyl)oxy)phenyl)acetoxy)ethyl)p¡peridin-1 -yl)ethyl)disulfan¡l)ethyl)piperidin-4-¡l)ethoxy)ethyl)phenyl) (Lipid 10). To a stirred solution of disulfide 14 (synthesis described in Example 9) (330 mg, 0.35 mmol) and acid 9c (synthesis described in Example 7 (143 mg, 0.39 mmol) in dichloromethane (20 mL) was added DMAP (47 mg, 0.39 mmol) followed by EDCI (60 mg, 0.39 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL) and dried over NasSCU. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography using 0-5% methanol in dichloromethane as eluent. The fraction containing the desired compound was evaporated to produce Lipid 10. (150 mg, 33%)1H NMR(300 MHz, d-chloroform): δ 7.26 (d, 4 H), 7.03 (d, 4 H), 4.86 (m, 1 H), 4 .05-4.11 (t, 6 H), 3.58 (s, 4 H), 2.80-2.90 (m, 8 H), 2.50-2.70 (m, 8 H) , 2.26-2.29 (m, 4 H), 1.92-1.99 (m, 4 H), 1.50-1.80 (m, 24 H), 1.16-1.40 (m, 46 H), 0.87 (t, 9 H). Example 11: Synthesis of 1-nonyl 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-((9-oxo -9-(pentadecan-7yloxy)nonanoyl)ox¡)phen¡l)acetoxy)ethyl)p¡perídin-1 -yl)ethyl)disulfan¡l)ethyl)piperidin-4-¡l)ethoxy) ethyl)phenyl)nonanedioate (Lipid 11) Lipid 11 103 Synthesis of 1-nonyl 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4-((9-oxo-9- (pentadecan-7yloxy)nonanoyl)oxy)phen¡l)acetoxy¡)ethyl)piper¡din-1 -yl)ethyl)disulfan¡l)ethyl)pipend¡n-4-yl)ethoxy)ethyl)phenyl) nonanedioate (Lipid 11). To a stirred solution of disulfide 14 (synthesis described in Example 9) (260 mg, 0.28 mmol) and acid 9d (synthesis described in Example 8) (122 mg, 0.3 mmol) in DCM (20 ml) DMAP (37 mg, 0.3 mmol) was added followed by EDCI (47 mg, 0.3 mmol). The resulting mixture was stirred at room temperature overnight, then washed with saturated sodium bicarbonate solution (20 mL), brine (20 mL) and dried over NaaSCX. The solvent was removed under reduced pressure and the residue was purified by Silica gel column chromatography using 0-5% MeOH in DCM as eluent. The fraction containing the desired compound was evaporated to produce Lipid 11 (110 mg, 30%). 4 H), 4.86 (m, 1 H), 4.05-4.11 (t, 6 H), 3.59 (s, 4 H), 2.80-2.90 (m, 8 H), 2.50-2.70 (m, 8 H), 2.27-2.29 (m, 4 H), 1.90-2.20 (t, 4 H), 1.50-1.82 (m, 24 H), 1.10-1.40 (m, 50 H), 0.87 (t, 9 H). The following Lipids 12-20 in Table 4 were prepared by similar procedures with the appropriate starting materials and other modifications that would be within the knowledge of the person of ordinary skill in the art. Table 4. MA / IZ / ZUZZ / UOU l OI 1-(heptadecan-9-yl) nonanedioate 9-(4-(2-(2-(1-(2-((2-(4-(2-(2- (4-(((9Z,12Z)octadeca-9,12-dienoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)pipehdin4-yl)ethoxy)-2-oxoethyl )phenyl) (Lipid 12) 104 MA / IZ / ¿U¿¿ / UO3 / 01 1 -(heptadecan-9-yl) 9-(4-(2-(2-(1-(2-((2-(4-(2-)) nonanedioate (2-(4-((8-(2octylcyclopropyl)octano¡l)oxy)phenyl)acetoxy¡)ethyl)piperidin-1¡I)ethyl)disuIfanoyl)et¡l)piperidin-4-yl) ethoxy)-2-oxoethyl)phenyl)ethyl (Lipid 13) 1-(heptadecan-9-yl)9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4(stearoyloxy))) nonanedioate phenyl))acetoxy¡)ethyl)p¡peridín-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4yl)ethoxy)ethyl)phenyl) (Lipid 14) or 1-(heptadecan-9-yl) 9-(4-(2-oxo-2-(2-(1-(2-((2-(4-(2-(2-(4 (undecane)) nonanedioate loxi)phen¡lo))acetoxy¡)et¡l)p¡per¡din-1 -¡l)ethyl)d¡sulfan¡l)ethyl)p¡períd¡n-4yl)ethoxy )ethyl)phenyl) (Lipid 15) or 1-(heptadecan-9-yl)9-(4-(2-(2-(1-(2-((2-(4-(2-(2-(4(nonanoyloxy)phenyl)acetoxy)) nonanedioate ethyl))piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)-2oxoethyl)phenyl) (Lipid 16) 105 MA / IZ / ¿U¿¿ / UO3 / ΟΊ 1-nonyl nonanedioate 9-(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4- ((9-((3-oct¡lundecyl)oxy)-9)oxononanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)2-oxoethyl )phenyl) (Lipid 17) 1 -(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-((7-(heptadecan-9-yloxy)) 9-nonyl nonanedioate) -7oxoheptanoyl)oxy))phenyl)acetoxy¡)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)2-oxoethyl)phenyl) (Lipid 18) 1-nonyl nonanedioate 9-(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-((9-((3-octylundecyl))ox) )-9) oxononanoyl)ox¡)phen¡l)acetoxy¡)ethyl)p¡per¡din-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy¡)2-oxoethyl)phenyl ) (Lipid 19) 1-nonyl nonanedioate 9-(4-(2-(2-(1 -(2-((2-(4-(2-(2-(4-((7-((3-octylundecyl))ox) )-7)oxoheptanoyl)oxy)phenyl)acetoxy)ethyl)piperidin-1 -yl)ethyl)disulfanoyl)ethyl)piperidin-4-yl)ethoxy)2-oxoethyl)phenyl) (Lipid 20) 106 Example 2: Preparation of Lipid Nanoparticles Lipid nanoparticles (LNPs) were prepared with a weight ratio of total lipids to ceDNA of approximately 10:1 to 30:1. In summary, an ionizable lipid of the present invention, a non-cationic lipid (e.g., distearoylphosphatidylcholine (DSPC)), a component for providing membrane integrity (such as an ester, e.g., cholesterol), and a conjugated lipid molecule ( such as a PEG-lipid, for example, 1-(monomethoxy-polyethylene glycol)-2,3dimyristoylglycerol, with a PEG average molecular weight of 2000 (PEG-DMG)), are solubilized in alcohol (for example, ethanol) at a molar ratio of, for example, 50:10:37:3 or 20:40:38:2. The ceDNA was diluted to a desired concentration in buffer solution. For example, ceDNA was diluted to a concentration of 0.1 mg / mL to 0.25 mg / mL in a buffer solution comprising sodium acetate, sodium acetate and magnesium chloride, citrate, melic acid or melic acid and sodium chloride. In one example, ceDNA was diluted to 0.2 mg / mL in 10 to 50 mM citrate buffer, pH 4. The alcoholic lipid solution was mixed with aqueous ceDNA solution by using, for example, syringe pumps or a impact jet mixer, in a ratio of approximately 1:5 to 1:3 (vol / vol) with total flow rates greater than 10 mL / min. In one example, the alcoholic lipid solution was mixed with aqueous ceDNA at a ratio of approximately 1:3 (vol / vol) with a flow rate of 12 mL / min. The alcohol was removed and the buffer was replaced with PBS by dialysis. Alternatively, buffers were replaced with PBS by using centrifuge tubes. Simultaneous alcohol removal and buffer exchange can be performed, for example, by dialysis or tangential flow filtration. The lipid nanoparticles obtained are filtered through a sterile 0.2 pm pore filter. In one study, illustrative ceDNA-comprising lipid nanoparticles were prepared by using a lipid solution comprising SS-OP, DSPC, cholesterol, and DMG-PEG2000 (molar ratio 50:10:37:3) as a control. In some examples, a target portion of tissue such as N-acetylgalactosamine (GalNAc) was included. A portion of GalNAc such as triantennary GalNAc (GalNAc3) or tetraantennary GalNAc (GalNAc4) can be synthesized as is known in the art (see documents, WO2017 / 084987 and WO2013 / 166121) and chemically conjugated with lipid or PEG as is known in the art ( see, Resen et al., J. Biol. Chem. (2001) “Determination of the Upper Size Limit for Uptake and Processing of Ligands by the Asialoglycoprotein Receptor on Hepatocytes in Vitro and in Vivo” 276:375577-37584). Aqueous solutions of ceDNA were prepared in buffered solutions. The lipid solution and ceDNA solution were mixed by using an internal procedure in a NanoAssembler at a total flow rate of 12 mL / min with a lipid to ceDNA ratio of 1:3 (v / v). IVIA / t / zuzz / uoy l OI 107 Table 2A: Administration of test material in Study A Group No. Animals per group LNP Treatment Dose Level (mg / kg) Dose Volume (mL / kg) Treatment Regimen End Time Point 1 5 PBS 0.25 5 Once on DayO, IV Day 7 2 5 LNP 1 3 5 LNP 2 4 5 LNP 3 5 5 LNP 4 6 5 LNP 5 7 5 LNP 6 8 5 LNP 7 9 5 LNP 8 10 5 LNP 9 11 5 LNP 10 12 5 LNP 11 13 5 LNP 12 MA / t / zuzz / uoy / 01 Num = Number; IV = intravenous; ROA = route of administration; LNP = lipid nanoparticle Table 2B: Test Material Administration in Study Group B No. Animals per Group LNP Treatment Dose Level (mg / kg) Dose Volume (mL / kg) Treatment Regimen End Time Point 14 5 PBS 0.25 5 A time on Day 0, IV Day 7 15 5 LNP 13 16 5 LNP 14 17 5 LNP 15 18 5 LNP 16 19 5 LNP 17 20 5 LNP 18 Num = Number; IV = intravenous; ROA = route of administration; LNP = lipid nanoparticle 108 Table 2C: Administration of test material in Study C Group No. Animals per group LNP Treatment Dose Level (mg / kg) Dose Volume (mL / kg) Treatment Regimen Final Time Point 21 5 PBS 0.25 5 Once on Day 0, IV Day 7 22 5 LNP 19 0.25 23 5 LNP 19 1 24 5 LNP 20 0.25 25 5 LNP 20 1 26 5 LNP 21 0.25 27 5 LNP 21 1 Num = Number; IV = intravenous; ROA = route of administration; LNP = lipid nanoparticle Table 2D: Administration of test material in Study D Group No. Animals per group LNP Treatment Dose Level (mg / kg) Dose Volume (mL / kg) Treatment Regimen End Time Point 28 5 PBS 0.25 5 Once on Day 0, IV Day 7 29 5 LNP 22 30 5 LNP 23 31 5 LNP 24 32 5 LNP 25 33 5 LNP 26 Num = Number; IV = intravenous; ROA = route of administration; LNP = lipid nanoparticle Table 2E: Administration of test material in Study E Group No. Animals per group LNP Treatment Dose Level (mg / kg) Dose Volume (mL / kg) Treatment Regimen End Time Point 34 5 PBS 0.25 5 Once on Day 0, IV Day 7 35 5 LNP 27 36 5 LNP 28 37 5 LNP 29 38 5 LNP 30 Num = Number; IV = intravenous; ROA = route of administration; LNP = lipid nanoparticle 109 Table 3A: Description of LNP Compositions in Study A LNP Components of LNP (molar ratio) PBS Not applicable *LNP 1 SS-OP : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.2 : 38.6 : 2.9 : 0.48) in melic acid *LNP 2 SS- OP: DOPC: Chol: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7: 7.2: 38.6: 2.9: 0.48) in malic acid LNP 3 Lipid 5: DOPC: chol: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7: 7.2 : 38.6 : 2.9 : 0.48) LNP 4 Lipid 2: DOPC: col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.2 : 38.6 : 2.9 : 0.48) LNP 5 Lipid 1: DOPC: col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.2 : 38.6 : 2.9 : 0.48) LNP 6 Lipid 3: DOPC: col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.2 : 38.6 : 2.9 : 0.48) LNP 7 SS-OP : DOPC : Chol : DSPE-PCB1-5 : DSPE-PEG2000-GalNAc4 (47.0 : 6.7 : 35.8 : 10.0 :0.50) LNP 8 SS-OP : DOPC : Chol : DSPE-PCB1-10 : DSPE-PEG2000-GalNAc4 (47.0 : 6.7 : 35.8 : 10.0 :0.50) LNP 9 SS-OP : DOPC : Chol : DSPE-PCB1-30 : DSPE-PEG2000-GalNAc4 (47.0 : 6.7 : 35.8 : 10.0 :0.50) LNP 10 SS-OP : DOPC : Chol : DSPE-PCB1-5 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.50) LNP 11 SS-OP : DOPC : Chol : DSPE-PCB1-10 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.50) LNP 12 SS-OP : DOPC : Chol : DSPE-PCB1-30 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.50) DOPC = dioleoylphosphatidylcholine; chole = cholesterol; DSPE = distearoyl-phosphatidyl-ethanolamine; DMG-PEG2000 = 1-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG2000-DMG); and SS-OP = COATSOME® SS-OP (NOF); GalNAc = N-acetylgalactosamine; GalNAc4 = Tetraantennary GalNAc *LNP1 and LNP2 contain the same components and the same molar ratio of the components, but they were manufactured in different batches and used as a control. 110 Table 3B: Description of LNP Compositions in Study B LNP LNP components (molar ratio) PBS Not applicable LNP 13 SS-OP : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 14 Lipid 4: DOPC: Chol: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 15 Lipid 5: DOPC: Col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 16 Lipid 2: DOPC: Col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 17 Lipid 1 : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 18 Lipid 3 : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) DOPC = dioleoylphosphatidylcholine; chole = cholesterol; DSPE = distearoyl-phosphatidyl-ethanolamine; DMGPEG2000 = 1-(monomethoxy¡-polyethylene glycol)-2,3-dimyristoylglycerol (PEG2000-DMG); and SS-OP = COATSOME® SS-OP (NOF); GalNAc = N-acetylgalactosamine; GalNAc4 = tetraantennary GalNAc Table 3C: Description of LNP Compositions in Study C LNP LNP components (molar ratio) PBS Not applicable LNP 19 SS-OP : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 20 Lipid 1 : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 21 Lipid 3 : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) DOPC = dioleoylphosphatidylcholine; chole = cholesterol; DSPE = distearoyl-phosphatidyl-ethanolamine; DMGPEG2000 = 1-(monomethoxy¡-polyethylenegl¡col)-2,3-d¡m¡r¡sto¡lgl¡cerol (PEG2000-DMG); and SS-OP = COATSOME® SS-OP (NOF); GalNAc = N-acetylgalactosamine; GalNAc4 = tetraantennary GalNAc 111 Table 3D: Description of LNP Compositions in Study D LNP LNP components (molar ratio) PBS Not applicable LNP 22 Onizable Lipid A: DOPC: Chol: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 23 SS-OP : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 24 Lipid 6 : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5 ) LNP 25 Lipid 7: DOPC: Chol: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7: 7.3: 38.6: 2.9: 0.5) LNP 26 Lipid 8: DOPC: Col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) DOPC = dioleoylphosphatidylcholine; chole = cholesterol; DSPE = distearoyl-phosphatidyl-ethanolamine; DMGMA / C / ZUZZ / UO3 / 01 PEG2000 = 1 -(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG2000-DMG); and SS-OP = COATSOME® SS-OP (NOF); GalNAc = N-acetylgalactosamine; GalNAc4 = tetraantennary GalNAc Table 3E: Description of LNP Compositions in Study E LNP LNP components (molar ratio) PBS Not applicable LNP 27 SS-OP : DOPC : Chol : DMG-PEG2000 : DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 28 Lipid 9: DOPC: Chol: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 29 Lipid 10: DOPC: Col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) LNP 30 Lipid 11: DOPC: Col: DMG-PEG2000: DSPE-PEG2000-GalNAc4 (50.7 : 7.3 : 38.6 : 2.9 : 0.5) DOPC = dioleoylphosphatidylcholine; chole = cholesterol; DSPE = distearoyl-phosphatidyl-ethanolamine; DMGPEG2000 = 1-(monomethoxy-polyethylene glycol)-2,3-dimyristo¡lglycerol (PEG2000-DMG); and SS-OP = COATSOME® SS-OP (NOF); GalNAc = N-acetylgalactosamine; GalNAc4 = tetraantennary GalNAc 112 Table 4: Blood collection Group Number Whole blood sample collection times (Tail, saphenous or orbital) SERUMa 1 -11 day 0 approximately 5 to 6 hours after dose of test material (not less than 5.0 hours, not more than 6.5 hours) Volume / per tion approximately 150 pL whole blood Processing / storage 1 aliquot frozen at nominally -70 °C aWhole blood was collected in serum separator tubes, with coagulation activator Species (number, sex, age): CD-1 mice (N = 65 and 5 spare, male, about 4 weeks old on arrival). Cage side observations: Cage side observations were performed daily. Clinical Observations: Clinical observations were made at approximately 1, approximately 5 to approximately 6 and approximately 24 hours after the day 0 test material dose. Additional observations were made by exception. Body weights were recorded for all animals, as appropriate, on days 0, 1, 2, 3, 4 & 7 (before euthanasia). Additional body weights were recorded as necessary. Dose administration: Test articles (LNP:ceDNA-Luc) were dosed at 5 mL / kg on day 0 for groups 1 to 38 by intravenous administration into the lateral tail vein. Live imaging: On day 4, all animals were dosed with luciferin at 150 mg / kg (60 mg / mL) via intraperitoneal (IP) injection at 2.5 mL / kg. <15 minutes after each luciferin administration; all animals had one IVIS imaging session according to the in vivo imaging protocol described below. Recovery from anesthesia: Animals were monitored continuously while under anesthesia, during recovery, and until they moved. Interim blood collection: All animals had provisional blood collected on day 0; 5-6 hours after dosing of test material (not less than 5.0 hours, not more than 6.5 hours). After each collection, the animals received 0.5 - 1.0 mL of Ringer's lactate, subcutaneously. 113 Whole blood for serum was collected by cutting the tail vein, saphenous vein, or orbital sinus (with inhalant isoflurane). Whole blood was collected in a serum separator tube, with coagulation activator and processed into one (1) aliquot of serum. In vivo imaging protocol. Luciferin stock powder was nominally stored at −20°C. • Formulated luciferin stored in 1 ml aliquots at 2 - 8°C protect from light. . The formulated luciferin was stable for up to 3 weeks at 2 - 8'C, protected from light and stable for about 12 hours at room temperature (RT). • Luciferin dissolved in PBS at a target concentration of 60 mg / mL at a sufficient volume and adjusted to pH = 7.4 with 5 M NaOH (approximately 0.5 pL / mg luciferin) and HCl (approximately 0.5 pL / mg luciferin) according to be necessary. . The appropriate amount was prepared according to the protocol, including approximately at least 50% surplus. Injection and Imaging (Note: Up to 5 animals can be imaged at a time) • The animal's hair cover was shaved (as necessary). . Per protocol, 150 mg / kg luciferin in PBS was injected at 60 mg / mL via IP. . Imaging was performed immediately or up to 15 minutes after the dose. . Adjust the isoflurane vaporizer to 1 - 3% (typically 2.5%) to anesthetize animals during imaging sessions. . Anesthesia with isoflurane for imaging session: Place the Animal in the isoflurane chamber and wait for the isoflurane to take effect, about 2-3 minutes. He made sure the anesthesia level on the side of the IVIS machine was in the on position. The animal(s) was placed in the IVIS machine The desired Acquisition Protocol was performed with settings for the highest sensitivity. Results I study As shown in Figure 1, on day 4, the group of mice treated with ceDNA-luciferase (ceDNA-luc) which were formulated with lipid 1, lipid 2, lipid 3 or lipid 5 (LNP 5, 4, 6 and 3 , respectively, from Figure 1) showed equivalent or higher luciferase expressions and / or activity compared to the positive control ceDNA LNP-treated groups used in Study A (LNP 1, 2 and 7-12, each which was a ceDNA-luc formulated with SS-OP lipids), suggesting that the ionizable lipids described herein possess superior physical attributes as a lipid nanoparticle delivery vehicle. ΜΛ / t / zuzz / uoy l OI 114 Study B As shown in Figure 2, and consistent with the observations in Figure 1 of Study A above, on Day 4, the group of mice treated with ceDNA-luc that were formulated with Lipid 1, Lipid 2, Lipid 3 or Lipid 5 (LNP 17, 16, 18 and 15, respectively, of Figure 2) exhibited equivalent or higher luciferase activity compared to that of the positive control ceDNA LNP-treated groups used in Study B (LNP 13 which was formulated with ceDNA-luc with lipid SS-OP), suggesting that the ionizable lipids described herein possess superior physical attributes as a lipid nanoparticle delivery vehicle. Studio C Lipids 1 and 3 that exhibited the highest luciferase expression and / or activity in Studies A and B were further studied in Study C to determine the dose response. As shown in Figure 3, and consistent with the observations of Figures 1 and 2 of Studies A and B, on Day 4 the group of mice treated with ceDNA-luc that were formulated with Lipid 1 or Lipid 3 (LNP 20 and 21, respectively, of Figure 1) exhibited higher luciferase expression and / or activity at both 25 mg / kg and 1 mg / kg compared to that of the positive control ceDNA LNP-treated groups used in Study C (LNP 19, which was a ceDNA-luc formulated with SSOP lipid), suggesting that the ionizable lipid described herein possess superior physical attributes as a lipid nanoparticle delivery vehicle. Furthermore, the results of Figure 3 indicate that LNP 20, when increased from a dose of 0.25 mg / kg to 1 mg / kg, exhibited a greater increase in luciferase expression and / or activity, compared to LNP 19 which was also tested at the same dosage levels. These results suggest that the LNPs formulated with the ionizable lipids of the present description are more sensitive to different dosage levels and that the level of expression of the transgenic insert in the ceDNA encapsulated by the LNPs formulated with the ionizable lipids of the present description, may be more easily adjusted to the level required to exert their therapeutic effect for a specific disease, demonstrating another convenient technical characteristic that these ionizable lipids possess as a delivery vehicle for lipid nanoparticles. Studio D In Study D, LNPs formulated with Lipid 6, Lipid 7, and Lipid 8 (LNP 24, 25, and 26, respectively from Figures 4A and 4B) and ceDNA-luc were evaluated for luciferase expression and / or activity in mice. and also the tolerability and were compared with LNPs formulated with ionizable lipid A and SS-OP lipid (LNP 22 and 23 respectively from Figures 4A and 4B) and ceDNA-luc, as shown in Figure 4A, on day 4, the group of mice treated with ceDNA-luc constructs that were formulated with lipid 6, lipid 7 and lipid 8 exhibited equivalent or higher luciferase expressions and / or activity compared to the groups treated with ceDNA-luc that was formulated with lipid SS- OP (i.e. LNP 23). Figure 4B indicates that the ceDNA-luc constructs MA / IZ / ¿U¿¿ / UO31 OI 115 formulated with lipid 6, lipid 7 and lipid 8 were also well tolerated in mice because the treatment did not cause changes in the body weight of the mice on Day 1. In contrast, as can be seen in Figure 4B, mice treated with ceDNA-luc formulated with ionizable lipid A (i.e., LNP 22) suffered significant weight loss on Day 1, indicating that the animals did not tolerate the lipid well. Study and In Study E, LNPs formulated with Lipid 9, Lipid 10 and Lipid 11 (LNP 28, 29 and 30, respectively from Figures 5A and 5B) and ceDNA-luc were evaluated for luciferase expression and / or activity in mice and also the tolerability and were compared with LNPs formulated with SS-OP lipid (LNP 27 of Figures 5A and 5B) and ceDNA-luc, as shown in Figure 5A, on day 4, the group of mice treated with constructs of ceDNA-luc that were formulated with lipid 9, lipid 10, and lipid 11 exhibited equivalent or higher luciferase expressions and / or activity compared to that of the groups treated with ceDNA-luc that was formulated with lipid SS-OP (i.e. LNP 27). Figure 5B indicates that, with the exception of one outlier data point at LNP 30, the ceDNA-luc constructs formulated with Lipid 9, Lipid 10, and Lipid 11 were generally well tolerated in mice because the treatment did not cause significant changes in the body weight in mice on Day 1. Therefore, Studies A-E generally demonstrate that LNPs formulated with the ionizable lipids of the present description: (i) have an excellent level of in vivo expression of the transgenic ceDNA insert; (i) they are sensitive to different dosage levels, thereby allowing the in vivo expression level of the transgenic ceDNA insert to be adjusted as necessary; and (iii) they are well tolerated in vivo. REFERENCES All publications and references, including, but not limited to, patents and patent applications, cited in this specification and in the Examples herein, are incorporated by reference in their entirety as if specifically and individually indicated that each publication or reference individual is hereby incorporated by reference as fully described. Any patent application to which this application claims priority is also incorporated herein by reference in the manner described above for publications and references.

Claims

1. A lipid having Formula (I): or a pharmaceutically acceptable salt thereof, wherein: a is an integer ranging from 1 to 20; b is an integer ranging from 2 to 10; R1 is absent or selected from alkenyl(C2-C2o), -C(O)Oalkyl(C2-C2o) and cyclopropyl substituted with alkyl(C2-C2o); and R2 is alkyl(C2-C2o); 2. The lipid according to claim 1, wherein the lipid is of Formula (II): or a pharmaceutically acceptable salt thereof, wherein cyd are each independently integers ranging from 1 to 8.

3. The lipid according to claim 2, or a pharmaceutically acceptable salt thereof, wherein cyd are each independently integers ranging from 2 to 8.

4. The lipid according to claim 2 or 3, or a pharmaceutically acceptable salt thereof, wherein cyd are each independently integers ranging from 4 to 8. 117 5. The lipid according to any one of claims 2 to 4, or a pharmaceutically acceptable salt thereof, wherein cyd are each independently integers ranging from 6 to 8.

6. The lipid according to claim 2, or a pharmaceutically acceptable salt thereof, wherein cyd are each independently 1, 3, 5 or 7.

7. The lipid according to any one of claims 2 to 6, or a pharmaceutically acceptable salt thereof, wherein at least one of cyd is 7.

8. The lipid according to any one of claims 1 to 7, wherein the lipid is of Formula (III): or a pharmaceutically acceptable salt thereof 9. The lipid according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein b is an integer ranging from 3 to 9.

10. The lipid according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein b is an integer ranging from 5 to 7.

11. The lipid according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein b is 5 or 7.

12. The lipid according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 2 to 18.

13. The lipid according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 3 to 17. 118 14. The lipid according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 6 to 18.

15. The lipid according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 4 to 12.

16. The lipid according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 2 to 5.

17. The lipid according to claim 16, or a pharmaceutically acceptable salt thereof, wherein a is 3.

18. The lipid according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 6 to 8.

19. The lipid according to claim 18, or a pharmaceutically acceptable salt thereof, wherein a is 7.

20. The lipid according to claim 18, or a pharmaceutically acceptable salt thereof, wherein a is 8.

21. The lipid according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 16 to 18.

22. The lipid according to claim 21, or a pharmaceutically acceptable salt thereof, wherein a is 17.

23. The lipid according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein a is an integer ranging from 9 to 11.

24. The lipid according to claim 23, or a pharmaceutically acceptable salt thereof, wherein a is 10.

25. The lipid according to any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein R1 is absent or selected from alkenyl(C5C15), -C(O)Oalkyl(C4-C18), and alkyl-substituted cyclopropyl(C4-C16). 119 26. The lipid according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, wherein R1 is absent or selected from alkenyl (C5C12), -C(O)Oalkyl(C4-C12) and alkyl-substituted cyclopropyl (C4-C12).

27. The lipid according to any one of claims 1 to 26, or a pharmaceutically acceptable salt thereof, wherein R1 is absent or selected from alkenyl (C5C10), -C(0)Oalkyl(C4-C10) and alkyl-substituted cyclopropyl (C4-C10).

28. The lipid according to any one of claims 1 to 27, or a pharmaceutically acceptable salt thereof, wherein R1 is a C10 alkenyl.

29. The lipid according to any one of claims 1 to 27, or a pharmaceutically acceptable salt thereof, wherein the alkyl at -C(0)Oalkyl(C2-C2o), C(O)Oalkyl(C4-Ci8), -C(O)Oalkyl(C4-Ci2), or -C(0)Oalkyl(C4-Cio) for R1 is an unbranched alkyl.

30. The lipid according to claim 29, or a pharmaceutically acceptable salt thereof, wherein R1 is -C(O)O(C9 alkyl).

31. The lipid according to any one of claims 25 to 27, or a pharmaceutically acceptable salt thereof, wherein the alkyl in C(0)Oalkyl(C4-Ci8), C(O)Oalkyl(C4-Ci2), or -C(0)Oalkyl(C4-Cio) is a branched alkyl.

32. The lipid according to claim 31, or a pharmaceutically acceptable salt thereof, wherein R1 is -C(O)O(C17 alkyl).

33. The lipid of any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from any group listed in Table 1.

34. The lipid according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R2 is selected from any group listed in Table 2.

35. The lipid according to claim 1, wherein the lipid is selected from any lipid listed in Table 3, or a pharmaceutically acceptable salt thereof. ma / t / zuzz / uoy 101 120 36. A lipid nanoparticle (LNP) comprising the lipid according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof; and a nucleic acid.

37. The lipid nanoparticle according to claim 36, wherein the nucleic acid is encapsulated in the lipid.

38. The lipid nanoparticle according to claim 36 or claim 37, wherein the nucleic acid is selected from the group consisting of minigenes, plasmids, minicircles, small interfering RNA (ipRNA), microRNA (miRNA), antisense oligonucleotides (ASO), nbozymes, cDNA, ministrand, doggybone™, protelomer closed-end DNA or dumbbell linear DNA), Dicer substrate dsRNA, small hairpin RNA (hpRNA), asymmetric interfering RNA (aRNA), microRNA (miRNA) viral vectors, mRNA, tRNA, rRNA, DNA, viral RNA vector, non-viral vector, and any combination thereof.

39. The lipid nanoparticle according to claim 38, wherein the nucleic acid is a closed-end DNA (cDNA).

40. The lipid nanoparticle according to any one of claims 36 to 39, further comprising a steral.

41. The lipid nanoparticle according to claim 40, wherein the steral is cholesterol or beta-sitosterol.

42. The lipid nanoparticle according to any of claims 36 to 41, further comprising a PEG-lipid conjugate.

43. The lipid nanoparticle according to claim 42, wherein the PEG-lipid conjugate is 1-(monomethoxy-polyethylene glycol)-2,3-dimethylstoylglycerol (PEG-DMG).

44. The lipid nanoparticle according to any of claims 36 to 43, further comprising a non-cationic lipid.

45. The lipid nanoparticle according to claim 44, wherein the non-cationic lipid is selected from the group consisting of distearoyl-sn-glycerophosphoethanolamine, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), MA. t / zuzz / uoy lOI 121 dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoylphosphatidylcholine (POPO), palmitoylphosphatidylethanolamine (POPE), dioleoylphosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), monomethyl-phosphatidylethanolamine (such as 16-O-monomethyl PE), dimethylphosphatidylethanolamine (such as 16-O-dimethyl PE), 18-1-trans PE, 1-stearoyl-2-oleoylphosphatidyethanolamine (SOPE), hydrogenated soy phosphatidylcholine (HSPC), egg phosphatidylcholine (EPC),dioleoylphosphatidylserine (DOPS), sphingomyelin (SM), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphatidylglycerol (DMPG), distearoylphosphatidylglycerol (DSPG), dierucoylphosphatidylcholine (DEPC), palmitoyloleiolphosphatidylglycerol (POPG), dielaidoyl-phosphatidylethanolamine (DEPE), 1,2-dilauroylsn-glycero-3-phosphoethanolamine (DLPE); 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (DPHyPE); lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetyl phosphate, lysophosphatidylcholine, dilinoleol-2-phosphatidylcholine, and mixtures thereof.

46. ​​The lipid nanoparticle according to claim 45, wherein the non-cationic lipid is selected from the group consisting of dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylcholine (DSPC), and dioleoylphosphatidylethanolamine (DOPE).

47. The lipid nanoparticle according to claim 46, wherein the PEG-lipid conjugate is present in a molar percentage of approximately 1.5% to approximately 4%.

48. The lipid nanoparticle according to claim 47, wherein the PEG-lipid conjugate is present in a molar percentage of approximately 2% to approximately 3%.

49. The lipid nanoparticle according to claim 48, wherein the PEG-lipid conjugate is present in a molar percentage of approximately 2.5 to approximately 3 o / o.

50. The lipid nanoparticle according to claim 49, wherein the PEG-lipid conjugate is present in a molar percentage of approximately 3%.

51. The lipid nanoparticle according to any one of claims 42 to 50, wherein the PEG-lipid conjugate is DMG-PEG. 122 52. The lipid nanoparticle according to any one of claims 36 to 51, wherein the cholesterol or beta-sitosterol is present in a molar percentage of approximately 20% to approximately 40%, and wherein the lipid is present in a molar percentage of approximately 80% to approximately 60%.

53. The lipid nanoparticle according to claim 52, wherein cholesterol or beta-sitosterol is present in a molar percentage of approximately 40%, and wherein the lipid is present in a molar percentage of approximately 50%.

54. The lipid nanoparticle according to any one of claims 36 to 39, further comprising a cholesterol, PEG-lipid conjugate and a non-cationic lipid.

55. The lipid nanoparticle according to claim 54, wherein the PEG-lipid conjugate is present in approximately 1.5% to approximately 4%.

56. The lipid nanoparticle according to claim 55, wherein the PEG-lipid conjugate is present in approximately 2% to approximately 3%.

57. The lipid nanoparticle according to claim 56, wherein the PEG-lipid conjugate is present in approximately 2.5% to approximately 3%.

58. The lipid nanoparticle according to claim 57, wherein the PEG-lipid conjugate is present in approximately 3%.

59. The lipid nanoparticle according to any one of claims 42 to 47, wherein cholesterol is present in a molar percentage of approximately 30% to approximately 50%.

60. The lipid nanoparticle of any one of claims 54 to 60, wherein the PEG-lipid conjugate is DMG-PEG2000.

61. The lipid nanoparticle according to any one of claims 53 to 60, wherein the lipid is present in a molar percentage of approximately 42.5% to approximately 62.5%.

62. The lipid nanoparticle according to any one of claims 53 to 60, wherein the non-cationic lipid is present in a molar percentage of approximately 2.5% to approximately 12.5%.

63. The lipid nanoparticle according to any one of claims 53 to 60, wherein cholesterol is present in a molar percentage of approximately 40%, the lipid is present in a molar percentage of approximately 52.5%, the non-cationic lipid is present in a molar percentage of approximately 7.5%, and wherein the PEG-lipid conjugate is present in approximately 3%.

64. The lipid nanoparticle according to any of claims 36 to 63, further comprising dexamethasone palmitate.

65. The lipid nanoparticle according to any one of claims 36 to 64, wherein the nanoparticle has a diameter ranging from approximately 50 nm to approximately 110 nm.

66. The lipid nanoparticle according to any one of claims 36 to 64, wherein the nanoparticle has a size less than approximately 100 nm.

67. The lipid nanoparticle according to claim 66 has a size smaller than approximately 70 nm, wherein the particle 68. The lipid nanoparticle according to claim 67 has a size smaller than approximately 60 nm, wherein the particle 69. The lipid nanoparticle according to claim 39, wherein the particle has a total lipid to cDNA ratio of approximately 10:

1.

70. The lipid nanoparticle according to claim 39, wherein the particle has a total lipid to cDNA ratio of approximately 20:

1.

71. The lipid nanoparticle according to claim 39, wherein the particle has a total lipid to cDNA ratio of approximately 30:

1. 124 72. The lipid nanoparticle according to claim 39, wherein the particle has a total lipid to cDNA ratio of approximately 40:

1.

73. The lipid nanoparticle according to any one of claims 36 to 72, further comprising a tissue-specific targeting portion.

74. The lipid nanoparticle according to claim 73, wherein the tissue-specific targeting portion is N-acetylgalactosamine (GalNAc); wherein GalNAc is bound to the PEG-lipid conjugate; and the GalNAc bound to the PEG-lipid conjugate is present in the particle in a molar percentage of approximately 1.5%, approximately 1.4%, approximately 1.3%, approximately 1.2%, approximately 1.1%, approximately 1.0%, approximately 0.9%, approximately 0.8%, approximately 0.7%, approximately 0.6%, approximately 0.5%, approximately 0.4%, approximately 0.3%, approximately 0.2%, or approximately 0.1%.

75. The lipid nanoparticle according to claim 74, wherein the PEG-lipid conjugate bound to GalNac is present in the particle in a molar percentage of approximately 0.5%.

76. The lipid nanoparticle according to any one of claims 36 to 75, further comprising approximately 10 mM to approximately 30 mM of malic acid.

77. The lipid nanoparticle according to claim 76, comprising approximately 20 mM of malic acid.

78. The lipid nanoparticle according to any one of claims 36 to 77, further comprising approximately 30 mM to approximately 50 mM of NaCl 79. The lipid nanoparticle according to claim 78, further comprising approximately 40 mM of NaCl 80. The lipid nanoparticle according to any one of claims 36 to 79, further comprising approximately 20 mM to approximately 100 mM of MgClz- 81. The lipid nanoparticle according to claim 39, wherein the cDNA is a linear closed-end duplex DNA. 125 82. The lipid nanoparticle according to claim 39, wherein the cDNA comprises an expression cassette, and wherein the expression cassette comprises a promoter sequence and a transgene.

83. The lipid nanoparticle according to claim 82, wherein the expression cassette comprises a polyadenylation sequence.

84. The lipid nanoparticle according to any one of claims 81 to 83, wherein the cDNA comprises at least one inverted terminal repeat (ITR) flanking the 5' or 3' end of said expression cassette.

85. The lipid nanoparticle according to claim 84, wherein the expression cassette is flanked by two ITRs, wherein the two ITRs comprise an ITR 5' and an ITR 3'.

86. The lipid nanoparticle according to claim 84, wherein the expression cassette is connected to an ITR at the 3' end (ITR 3j).

87. The lipid nanoparticle according to claim 84, wherein the expression cassette is connected to an ITR at the 5' end (ITR 5j).

88. The lipid nanoparticle according to claim 84, wherein at least one of the ITR 5' and ITR 3' is a wild-type AAV ITR.

89. The lipid nanoparticle according to claim 84, wherein at least one of ITR 5' and ITR 3' is a modified ITR.

90. The lipid nanoparticle according to claim 84, wherein the cDNA further comprises a spade sequence between a 5' ITR and the expression cassette.

91. The lipid nanoparticle according to claim 84, wherein the cDNA further comprises a spade sequence between a 3' ITR and the expression cassette.

92. The lipid nanoparticle according to claim 90 or claim 91, wherein the sword sequence has a length of at least 5 base pairs.

93. The lipid nanoparticle according to claim 92, wherein the sword sequence has a length of 5 to 100 base pairs. MA / IZ / ¿U¿¿ / UO31 OI 126 94. The lipid nanoparticle according to claim 92, wherein the sword sequence has a length of 5 to 500 base pairs.

95. The lipid nanoparticle according to any one of claims 38 to 94, wherein the DNAce has a notch or an interruption.

96. The lipid nanoparticle according to claim 84, wherein the ITR is an ITR derived from an AAV serotype, derived from a goose virus ITR, derived from a B19 virus ITR, a wild-type parvovirus ITR.

97. The lipid nanoparticle according to claim 96, wherein said AAV serotype is selected from the group comprising AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11 and AAV12.

98. The lipid nanoparticle according to claim 84, wherein the ITR is a mutating ITR, and the cDNA optionally comprises an additional ITR that differs from the first ITR.

99. The lipid nanoparticle according to claim 84, wherein the cDNA comprises two mutant ITRs at the 5' and 3' ends of the expression cassette, wherein optionally the two mutant ITRs are symmetric mutants.

100. The lipid nanoparticle according to claim 39, wherein the DNAce is a CELiD, a DNA-based minicircle, a MIDGE, a DNA ministrand, a dumbbell-shaped linear closed-end duplex DNA comprising two ITR hairpin structures at the 5' and 3' ends of an expression cassette, or a doggybone™ DNA.

101. A pharmaceutical composition comprising the lipid nanoparticle according to any of claims 36 to 100 and a pharmaceutically acceptable excipient.

102. A pharmaceutical composition comprising the lipid according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable excipient.

103. A method for treating a genetic disorder in a subject, the method comprising administering to the subject an effective amount of the lipid nanoparticle according to any one of claims 36 to 100, or an effective amount of the pharmaceutical composition according to claim 101.

104. The method according to claim 103, wherein the subject is a human.

105. The method according to claim 103 or claim 104, wherein the genetic disorder is selected from the group consisting of sickle cell anemia, melanoma, hemophilia A (coagulation factor VIII (FVIII) deficiency) and hemophilia B (coagulation factor IX (FIX) deficiency), cystic fibrosis (CFTR), familial hypercholesterolemia (LDL receptor defect), hepatoblastoma, Wilson's disease, phenylketonuria (PKU), congenital hepatic porphyria, hereditary disorders of liver metabolism, Lesch-Nyhan syndrome, sickle cell anemia, thalassemias, xeroderma pigmentosum, Fanconi anemia, retinitis pigmentosa, ataxia telangiectasia, Bloom syndrome, retinoblastoma, mucopolysaccharide storage diseases (e.g., Hurler syndrome (MPS type I), Scheie syndrome (MPS type IS), syndrome of Hurler-Scheie (MPS type I HS), Hunter syndrome (MPS type II), Sanfilippo types A, B, C and D (MPS types III A, B, C and D),Morquio types A and B (MPS IVA and MPS IVB), Maroteaux-Lamy syndrome (MPS type VI), Sly syndrome (MPS type VII), hyaluronidase defibrillation (MPS type IX), Niemann-Pick disease types A / B, C1 and C2, Fabry disease, Schindler disease, GM2 gangliosidosis type II (Sandhoff disease), Tay-Sachs disease, metachromatic leukodystrophy, Krabbe disease, mucolipidosis types I, II / III and IV, sialidosis types I and II, glycogen storage disease types I and II (Pompe disease), Gaucher disease types I, II and III, Fabry disease, cystinosis, Batten disease, aspartylglucosaminuria, Salla disease, Danon disease (LAMP-2 deficiency), lysosomal acid lipase (LAL) deficiency neuronal ceroid lipofuscinosis (CLN1-8, INCL and LINCL), sphingolipidosis, galactosialidosis, amyotrophic lateral sclerosis (ALS), Parkinson's disease, Alzheimer's disease, Huntington's disease, spinocerebellar ataxia,spinal muscular atrophy, Friedreich's ataxia, Duchenne muscular dystrophy (DMD), Becker muscular dystrophies (BMD), dystrophic epidermolysis bullosa (DEB), ectonucleotide pyrophosphatase 1 deficiency, generalized arterial calcification of childhood (GACI), Leber congenital amaurosis, Stargardt macular dystrophy (ABCA4), ornithine transcarbamylase (OTO) deficiency, Usher syndrome, alpha-1 antitrypsin deficiency, progressive familial intrahepatic cholestasis (PFIC) type I (ATP8B1 deficiency), type II (ABCB11), type III (ABCB4), or type IV (TJP2) and cathepsin A deficiency, 106. The method according to claim 105, wherein the genetic disorder is Leber congenital amaurosis (LCA). 128 107. The method according to claim 106, wherein LCA is LCA10.

108. The method according to claim 105, wherein the genetic disorder is Niemann-Pick disease.

109. The method according to claim 105, wherein the genetic disorder is Stargardt macular dystrophy.

110. The method according to claim 105, wherein the genetic disorder is glucose-6-phosphatase (G6Pase) deficiency (glycogen storage disease type I) or Pompe disease (glycogen storage disease type II).

111. The method according to claim 105, wherein the genetic disorder is hemophilia A (factor VIII deficiency).

112. The method according to claim 105, wherein the genetic disorder is hemophilia B (factor IX deficiency).

113. The method according to claim 105, wherein the genetic disorder is Hunter syndrome (Mucopolysaccharidosis II).

114. The method according to claim 105, wherein the genetic disorder is cystic fibrosis.

115. The method according to claim 105, wherein the genetic disorder is dystrophic epidermolysis bullosa (DEB).

116. The method according to claim 105, wherein the genetic disorder is phenylketonuria (PKU).

117. The method according to claim 105, wherein the genetic disorder is progressive familial intrahepatic cholestasis (PFIC).

118. The method according to claim 105, wherein the genetic disorder is Wilson's disease. 129 119. The method according to claim 105, wherein the genetic disorder is Gaucher disease type I, II or III.