Genetic Disorder Constructs

Abstract

Disclosed herein are polynucleotides, expression constructs, compositions, kits, and methods related to genetic disorders. Some embodiments provided herein encompass protein variants related to genetic disorders. Expression constructs encoding highly functional and / or stable protein variants are also contemplated. Also described herein are expression constructs encoding protein variants with reduced activity and / or stability. Also described herein are compositions, kits, and methods for diagnosing and treating genetic disorders. In some embodiments, the variants may be related to the treatment and / or diagnosis of genetic disorders.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 382,438, filed November 4, 2022, the entire contents of which are incorporated herein by reference.

[0002] Statement Regarding Federally Sponsored R&D This invention was made with government support under Grant No. R01 GM134274 awarded by the National Institutes of Health / National Institute of General Medical Sciences. The government has certain rights in this invention.

[0003] The present disclosure relates to polynucleotides, expression constructs, compositions, kits, and methods relating to genetic disorders.

[0004] Sequence Listing Reference This application is filed with an electronic Sequence Listing. The Sequence Listing is provided under the file name PNDRI.015WO.xml, created on October 25, 2023, and is 1,899,795 bytes in size. The information in the electronic Sequence Listing is incorporated herein by reference in its entirety. [Background technology]

[0005] Inborn errors of metabolism (IEM) are a class of genetic disorders that, if left untreated, can lead to debilitating conditions. These disorders represent a wide range of phenotypes, including lethality, severe neurological symptoms, seizures, and intellectual disability. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] U.S. Patent No. 5,399,346 [Patent Document 2] U.S. Patent No. 5,580,859 [Non-patent literature]

[0007] [Non-Patent Document 1] Shen et al., "Juvenile-onset PSAT1-related neuropathy: A milder phenotype of serine deficiency disorder," Front Genet., August 16, 2022; Vol. 13: 949038 (PMID: 36061210) Summary of the Invention [Means for solving the problem]

[0008] The present application relates to protein variants of PSAT1 having different amino acid substitutions. Any feature, structure, or step disclosed herein can be substituted, combined, or omitted with any other feature, structure, or step disclosed herein. Furthermore, certain aspects, advantages, and features of the present invention are described herein for the purpose of summarizing the disclosure. It should be understood that not all or any such advantages will necessarily be achieved in accordance with any particular embodiment of the invention disclosed herein. No individual aspect of this disclosure is essential or required.

[0009] In some embodiments, a protein having an amino acid sequence of any one of SEQ ID NOs: 1408-1537 is provided. In some embodiments, a protein at least 80% identical to SEQ ID NO: 1 (WT sequence) is provided, comprising one or more amino acid substitution mutations in Table 2. In some embodiments, a protein having at least one amino acid substitution mutation in Table 2 exhibits greater activity compared to the WT protein of SEQ ID NO: 1. In some embodiments, the function of a protein having at least one amino acid substitution mutation in Table 2 is enhanced by more than 5% compared to the function of the WT protein of SEQ ID NO: 1. In some embodiments, the function of a protein having at least one amino acid substitution mutation in Table 2 is enhanced by at least 10% compared to the function of the WT protein of SEQ ID NO: 1.

[0010] In some embodiments, the amino acid substitution variants are 2, 3, 4, 5, 7, 8, 12, 15, 17, 22, 24, 26, 29, 31, 37, 42, 46, 48, 49, 53, 55, 56, 63, 68, 71, 72, 73, 74, 80, 85, 93, 94, 95, 97, 106, 109, 110, 116, 117, 121, 125, 132, 133, 136, 138, 144, 146, 148, 157, 169, 174, 180, 193, 194, 195, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 226, 229, 331, 332, 333, 334, 335, 336, 337, 338, 340, 341, 342, 343, 344, 345, 346, 348, 357, 158, 169, 174, 175, 176, 177, 178, 179, 180, and at least one amino acid change at an amino acid position of SEQ ID NO: 1 selected from the group consisting of 98, 203, 219, 221, 226, 229, 231, 236, 239, 240, 242, 243, 245, 249, 250, 253, 255, 259, 284, 300, 302, 310, 311, 318, 325, 334, 336, 341, 344, 349, 350, 351, 353, 354, 356, 357, 359, and 369. In some embodiments, the amino acid substitution variants are 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 49 , 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 10 0, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 132, 133, 134, 135, 136, 137, 138, 139, 140 , 141, 142, 143, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 227, 228, 229, 230, 231 , 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281 , 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331 , 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370,

[0011] In some embodiments, an expression vector is provided comprising a polynucleotide sequence encoding any of the embodiments of the present application. In some embodiments, an expression vector is provided comprising a polynucleotide sequence encoding one or more proteins of SEQ ID NOs: 1408-1537. In some embodiments, the sequence is codon-optimized. In some embodiments, a polynucleotide sequence comprising one or more of the amino acid substitution mutations in Table 2 enhances function by more than 5% compared to the wild-type protein of SEQ ID NO: 1. In some embodiments, the protein is phosphoserine aminotransferase (PSAT1).

[0012] In some embodiments, a composition is provided comprising a protein of any one of SEQ ID NOs: 1408-1537. In some embodiments, the composition is for use in treating a genetic disorder. In some embodiments, the composition is administered to a subject having a genetic disorder in an amount effective to correct the genetic disorder or a symptom thereof in the subject. In some embodiments, the genetic disorder is phosphoserine aminotransferase (PSAT1) deficiency (PSATD). In some embodiments, the genetic disorder is Neuraxova syndrome 2 (NLS2). In some embodiments, the PSATD or a symptom thereof is reduced after administration. In some embodiments, the NLS2 or a symptom thereof is reduced after administration. In some embodiments, the subject is a fetus, newborn, juvenile, or adult. In some embodiments, one or more expression vectors are administered to the subject in an amount effective to correct the PSATD or a symptom thereof in the subject. In some embodiments, one or more expression vectors are administered to the subject in an amount effective to correct the NLS2 or a symptom thereof in the subject. In some embodiments, the PSAT1 is modified to exhibit enhanced function. In some embodiments, PSAT1 is replaced or complemented with one or more polynucleotide sequences encoding SEQ ID NOs: 4-1537 to exhibit enhanced function.

[0013] In some embodiments, the function of a protein having at least one amino acid substitution mutation in Table 2 is enhanced by more than 5% compared to the function of the WT protein of SEQ ID NO: 1. In some embodiments, the function of a protein having at least one amino acid substitution mutation in Table 2 is enhanced by at least 50% compared to the function of the WT protein of SEQ ID NO: 1.

[0014] In some embodiments, kits for determining specific treatment modes for genetic disorders are provided, the kits comprising a lookup table indicating specific amino acid substitutions commonly involved in cellular mechanisms and one or more vectors comprising the sequences of SEQ ID NOs: 4-1537. In some embodiments, the lookup table comprises the list of mutations in Table 2 and Table 3. In some embodiments of the kit, the genetic disorder is PSATD. In some embodiments of the kit, the genetic disorder is NLS2. In some embodiments, the lookup table comprises the list of mutations in Table 3 (which show decreased activity and are needed to increase activity). In some embodiments, the lookup table comprises the list of mutations in Table 2 (which can be used to identify sequences useful for enhancing activity).

[0015] In some embodiments, methods are provided for identifying a subject with a genetic disorder, comprising detecting the presence of at least one mutation in a protein that differs in a subject with the genetic disorder compared to a subject without the disorder. In some embodiments, the presence of the at least one mutation indicates the presence of a genetic disorder in the subject. In some embodiments, the genetic disorder or its symptoms are reduced after administration. In some embodiments, the mutation is selected from the group consisting of the mutations listed in Table 3. In some embodiments, the genetic disorder is PSATD. In some embodiments, the one mutation that indicates the presence of a genetic disorder in the subject is selected from the mutations in Table 3. In some embodiments, the genetically engineered protein administered to the subject comprises one or more of the mutations in Table 2.

[0016] In some embodiments, a method for ameliorating, alleviating, or treating a genetic disorder is provided, comprising: receiving diagnostic information comprising the detection of the presence of a DNA variant that results in at least one mutation in a protein that differs in a subject having the genetic disorder compared to a subject not having the genetic disorder, wherein the presence of the at least one mutation indicates the presence of the genetic disorder in the subject; and administering a therapeutic agent to the subject, wherein the genetic disorder or its symptoms are reduced after administration. In some embodiments, the therapeutic agent comprises an effective amount of a dietary supplement.

[0017] In some embodiments, the dietary supplement comprises L-serine. In some embodiments, an effective amount comprises 100-950 mg / kg / day of L-serine. In some embodiments, an effective amount comprises 400-950 mg / kg / day of L-serine. In some embodiments, an effective amount comprises 500 mg / kg / day of L-serine. In some embodiments, the dietary supplement comprises glycine. In some embodiments, an effective amount comprises 200 mg / kg / day of glycine. In some embodiments, an effective amount comprises 400 mg / kg / day of glycine. In some embodiments, an effective amount comprises 200-400 mg / kg / day of glycine. In some embodiments, the dietary supplement comprises L-serine and glycine. In some embodiments, an effective amount comprises 500 mg / kg / day of L-serine and 200 mg / kg / day of glycine. In some embodiments, an effective amount comprises 300 mg / kg / day of L-serine and 400 mg / kg / day of glycine. In some embodiments, the effective amount comprises 300-500 mg / kg / day of L-serine and 200-400 mg / kg / day of glycine. In some embodiments, the genetic disorder is PSATD. In some embodiments, the genetic disorder is NLS2. In some embodiments, the one mutation indicative of the presence of a genetic disorder in the subject is selected from the mutations in Table 3.

[0018] In some embodiments, a method for ameliorating, alleviating, or treating a disorder is provided, comprising: receiving diagnostic information comprising the detection of the presence of a DNA variant resulting in at least one mutation in a protein that differs in a subject having the disorder compared to a subject not having the disorder, wherein the presence of the at least one mutation indicates the presence of the disorder in the subject; and administering a therapeutic agent to the subject, wherein the disorder or its symptoms are reduced after administration, wherein the disorder is associated with reduced activity of serine biosynthesis. In some embodiments, the disorder comprises ichthyosis. In some embodiments, the disorder comprises epilepsy. In some embodiments, the disorder comprises hypertension. In some embodiments, the disorder comprises retinal degeneration. In some embodiments, the disorder comprises type 2 macular telangiectasia. In some embodiments, the mutant protein comprises an amino acid substitution of at least one amino acid in WT PSAT1. In some embodiments, the therapeutic agent comprises an effective amount of a dietary supplement.

[0019] In some embodiments, the dietary supplement includes L-serine. In some embodiments, an effective amount includes 100-950 mg / kg / day of L-serine. In some embodiments, an effective amount includes 400-950 mg / kg / day of L-serine. In some embodiments, an effective amount includes 500 mg / kg / day of L-serine. In some embodiments, the dietary supplement includes glycine. In some embodiments, an effective amount includes 200 mg / kg / day of glycine. In some embodiments, an effective amount includes 400 mg / kg / day of glycine. In some embodiments, an effective amount includes 200-400 mg / kg / day of glycine. In some embodiments, the dietary supplement includes L-serine and glycine. In some embodiments, an effective amount includes 500 mg / kg / day of L-serine and 200 mg / kg / day of glycine. In some embodiments, an effective amount includes 300 mg / kg / day of L-serine and 400 mg / kg / day of glycine. In some embodiments, the effective amount comprises 300-500 mg / kg / day of L-serine and 200-400 mg / kg / day of glycine.

[0020] The foregoing and other features of the present disclosure will become more fully apparent from the following description taken in conjunction with the accompanying drawings, in which: The present disclosure will be described with additional specificity and detail using the accompanying drawings, with the understanding that these drawings illustrate only some embodiments in accordance with the present disclosure and are therefore not to be considered limiting of its scope. [Brief explanation of the drawings]

[0021] [Figure 1] FIG. 1 shows the sequence of human PSAT1 as shown in GenBank accession number NP_478059.1. [Figure 2] FIG. 1 shows the sequence of the human PSAT1 protein, with amino acid positions in the sequence that can be substituted indicated by an X. [Figure 3] 1 is a diagram showing the human PSAT1 protein, with amino acid positions in the sequence that can be substituted indicated by an X. [Figure 4] FIG. 1 shows the sequences of human PSAT1 protein variants represented by SEQ ID NOs: 3 to 1407. [Figure 5] FIG. 1 shows the sequences of human PSAT1 protein variants represented by SEQ ID NOs: 1408 to 1537. [Figure 6] FIG. 1 shows the sequence of the expression construct shown in GenBank accession number MN654101. [Figure 7] 1 shows the results of a quantitative yeast assay of PSAT1 function in the absence of serine. Growth in the absence of exogenously supplied serine for a wild-type strain (codon-optimized human PSAT1 gene) and strains harboring amino acid substitution mutations of human PSAT1. Growth values ​​(y-axis) are plotted as points on a scatter plot in order of amino acid position (x-axis). [Figure 8]This figure shows the results of dietary supplementation across a variety of cases in treating symptoms manifested by at least one mutation in the human PSAT1 gene (reprinted from Shen et al., "Juvenile-onset PSAT1-related neuropathy: A milder phenotype of serine deficiency disorder," Front Genet., August 16, 2022; Vol. 13:949038 (PMID: 36061210)). DETAILED DESCRIPTION OF THE INVENTION

[0022] Dietary restrictions or supplements can have a positive impact on patient health, but timely recognition and targeted treatment are important. However, while IEMs share many common symptoms, they require fundamentally different treatments. Serine deficiency is an example of a highly actionable IEM. Serine plays a central role in processes such as cell proliferation, folate metabolism, phospholipid metabolism, and the formation of the neuromodulators D-serine and glycine. Serine deficiency can have significant effects on central nervous system development. PSAT1 deficiency is caused by mutations in the gene encoding phosphoserine aminotransferase (PSAT1), the second enzyme in the serine biosynthetic pathway. This rare but highly actionable disorder can lead to severe conditions such as Neuraxova syndrome 2, which is characterized by a recognizable pattern of severe malformations leading to prenatal or postnatal lethality.

[0023] Existing biochemical assays for PSAT1 deficiency have limitations, including the assessment of serine levels, which generally requires invasive measurement of metabolite concentrations in cerebrospinal fluid and comparison with closely age-matched controls. Computational methods for predicting the impact of variants derived from polymorphisms often yield inaccurate or contradictory results. Therefore, DNA sequencing-based diagnosis has the potential not only to reduce the morbidity and mortality associated with delayed treatment or underdiagnosis of various diseases, including IEM. However, because approximately 25% of PSAT1 deficiency cases occur de novo and only a small number of PSAT1 variants are recurrent, DNA sequencing often yields novel sequence variants of unknown significance (VUS).

[0024] The current existing biochemical assays for detecting PSAT1 deficiency (PSATD) are enzyme assays. These assays have limitations, including their sensitivity and specificity, which are used in newborn screening panels. In addition, only a small number of PSAT1 variants are recurrent, and DNA sequencing-based diagnosis often yields novel sequence variants of uncertain significance (VUS).

[0025] Identification of protein variants with specific attributes can be used to stratify patients for specific therapeutic agents. At present, it is not possible to computationally predict the functional consequences of all amino acid substitution mutations, so the results of functional assays such as those described herein for PSAT1 are required to accurately and quantitatively determine the extent to which an amino acid substitution will (or will not) alter protein activity, stability, or both.

[0026] Provided herein are protein variants of PSAT1. In some embodiments, they can be for applications in which a protein variant with increased activity and / or stability is desired. Also disclosed herein are protein variants with reduced activity and / or stability that can be used for diagnosing PSATD. In some embodiments, a protein variant with increased activity and / or stability can be used to replace or complement a protein variant with reduced activity and / or stability. In some embodiments, this identification can then be used to treat a subject as provided herein (e.g., by increasing the activity and / or function of PSAT1).

[0027] In some embodiments, a method for ameliorating, alleviating, or treating a genetic disorder is provided. The method comprises detecting the presence of at least one mutation in a subject having the genetic disorder compared to a subject not having the disorder, the mutation resulting in a protein variant with reduced activity and / or stability, wherein the presence of the at least one mutation resulting in a protein variant with reduced activity and / or stability indicates the presence of the genetic disorder in the subject. The method comprises administering to the subject a composition comprising at least one protein variant with increased activity and / or stability, wherein after administration, the genetic disorder or a symptom thereof is reduced.

[0028] term Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0029] As used herein, "a" or "an" may mean one or more than one.

[0030] "About," when used herein in reference to a measurable value, is meant to encompass a variation of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and even more preferably ±0.1% from the specified value.

[0031] Conditional language such as "can," "could," "could," "might," and the like, unless expressly stated otherwise or understood otherwise within the context in which it is used, is generally intended to convey that certain embodiments include certain features, elements, and / or steps, and other embodiments do not. Thus, such conditional language is generally not intended in any way to imply that features, elements, and / or steps are required for one or more embodiments.

[0032] Terms such as "comprising," "including," and "having" are synonymous and are used in an inclusive, open-ended manner and do not exclude additional elements, features, acts, operations, etc. Also, the term "or" is used in an inclusive (rather than exclusive) sense, so that, for example, when used to connect a list of elements, the term "or" means one, some, or all of the elements in the list.

[0033] Additionally, ranges disclosed herein include all overlaps, subranges, and combinations thereof. Terms such as "up to," "at least," "greater than," "less than," and "between" are inclusive of the recited values.

[0034] As used herein, numerical values ​​preceded by terms such as "approximately," "about," and "substantially" include the recited numerical value (e.g., about 10% = 10%) and represent an amount that approximates the stated amount that still performs the desired function and achieves the desired result. For example, the terms "approximately," "about," and "substantially" can refer to an amount that is within less than 10%, less than 5%, less than 1%, less than 0.1%, and less than 0.01% of the stated amount.

[0035] As used herein, "polynucleotide" refers to a "nucleic acid" or "nucleic acid molecule," e.g., deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), oligonucleotides, fragments produced by polymerase chain reaction (PCR), and fragments produced by ligation, cleavage, endonuclease action, and exonuclease action. Nucleic acid molecules can be composed of monomers that are natural nucleotides (e.g., DNA and RNA) or analogs of natural nucleotides (e.g., enantiomeric forms of natural nucleotides), or combinations of both. Modified nucleotides can have alterations in the sugar moiety and / or pyrimidine or purine base moieties. Sugar modifications include, for example, replacing one or more hydroxyl groups with halogens, alkyl groups, amines, and azide groups, or the sugar can be functionalized as an ether or ester. Additionally, the entire sugar moiety can be replaced with sterically and electronically similar structures, such as aza-sugars and carbocyclic sugar analogs. Examples of modifications at the base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well-known heterocyclic substituents. Nucleic acid monomers may be linked by phosphodiester bonds or analogs of such linkages. Phosphodiester linkage analogs include phosphorothioates, phosphorodithioates, phosphoroselenoates, phosphorodiselenoates, phosphoroanilothioates, phosphoranilidates, and phosphoramidates. The term "nucleic acid molecule" also includes so-called "peptide nucleic acids," which contain natural or modified nucleobases attached to a polyamide backbone. Nucleic acids may be single-stranded or double-stranded. In some alternative embodiments, nucleic acid sequences encoding fusion proteins are provided. In some alternative embodiments, the nucleic acid is RNA or DNA.

[0036] The terms "peptide," "polypeptide," and "protein," as used herein, have their plain and ordinary meaning as understood in light of the present specification and refer to polymers composed of amino acids linked by peptide bonds. Numerous functions of peptides, polypeptides, and proteins are known in the art, including, but not limited to, enzymatic, structural, transport, defensive, hormonal, or signal transduction. Peptides, polypeptides, and proteins are often produced biologically by ribosomal complexes using nucleic acid templates, although this is not always the case; chemical synthesis can also be used. By manipulating the nucleic acid template, peptide, polypeptide, and protein mutations can be made, such as substitutions, deletions, truncations, additions, duplications, or fusions of more than one peptide, polypeptide, or protein.Such fusions of more than one peptide, polypeptide, or protein may be adjacent in the same molecule or may have additional amino acids between them, e.g., linkers, repeats, epitopes, or tags, or may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 residues in length, or may be about 1, 2, 3, 4, 5 , 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 residues in length, , or 300 residues in length, or at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 residues in length It may be joined via any other sequence that is up to 75, 80, 85, 90, 95, 100, 150, 200, or 300 residues in length, or up to about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 residues in length, or any length in the range defined by any two of the foregoing lengths.

[0037] As used herein, a "highly active variant" refers to a protein that has at least one amino acid substitution and has increased activity compared to the wild-type or naturally occurring protein. A highly active variant exhibits enhanced functionality, abundance, activity, specificity, affinity, or a combination of the aforementioned parameters compared to the wild-type or naturally occurring protein.

[0038] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, gamma-carboxyglutamate, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an alpha carbon bonded to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refer to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that function in a manner similar to a naturally occurring amino acid.

[0039] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, gamma-carboxyglutamate, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an alpha carbon bonded to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refer to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that function in a manner similar to a naturally occurring amino acid.

[0040] As used herein, an "expression construct" or "construct" or "expression vector" or "vector" is a nucleic acid used to introduce a heterologous nucleic acid into a cell that contains regulatory elements that result in expression of the heterologous nucleic acid in the cell. Typically, an expression vector contains a transcription promoter, a gene, and a transcription terminator. Gene expression is usually placed under the control of a promoter, and such a gene is said to be "operably linked" to the promoter. Similarly, a regulatory element and a core promoter are operably linked if the regulatory element modulates the activity of the core promoter. Constructs described herein include, but are not limited to, plasmids, minicircles, yeast, and viral genomes.

[0041] A single copy of the construct is stably inserted into the yeast genome (either at the same location as the deleted yeast gene or at a neutral location within the genome, such as the HO locus). In embodiments herein, the construct is a "linear DNA molecule (double-stranded or single-stranded)" that can be integrated into the yeast nuclear or mitochondrial genome by homologous recombination. Such constructs can be synthesized in vitro by commercial gene synthesis companies (e.g., Twist Biosciences or IDT). Thus, the nucleic acid sequence can be a human sequence or a yeast-optimized version that encodes the same protein sequence.

[0042] "Encode" or "encode" have their plain and ordinary meaning when read in light of this specification and can include, but are not limited to, the property of a particular sequence of nucleotides in a polynucleotide, such as a gene, cDNA, or mRNA, to serve as a template for the synthesis of another polymeric molecule, such as a defined amino acid sequence. Thus, a gene encodes a protein when transcription and translation of mRNA corresponding to the gene produces the protein in a cell or other biological system.

[0043] The terms "identical" or percent "identity" in the context of two or more nucleic acid or polypeptide sequences refer to two or more sequences or subsequences that are the same sequence. Two sequences are "substantially identical" if the two sequences have a specified percentage of the same amino acid residues or nucleotides (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity over a specified region, or, if not specified, over the entire sequence of a reference sequence), when compared and aligned for maximum correspondence using one of the sequence comparison algorithms below or by manual alignment and visual inspection over a comparison window, or designated region. Some embodiments provided herein provide polypeptides or polynucleotides that are substantially identical to the polypeptides or polynucleotides, respectively, exemplified herein. Optionally, identity exists over a region that is at least about 15, 25, or 50 nucleotides in length, or more preferably over a region that is 100-500 or 1000 or more nucleotides in length, or over the entire length of the reference sequence. With respect to amino acid sequences, identity or substantial identity may exist over a region that is at least 5, 10, 15, or 20 amino acids in length, optionally at least about 25, 30, 35, 40, 50, 75, or 100 amino acids in length, optionally at least about 150, 200, or 250 amino acids in length, or the entire length of the reference sequence. With respect to shorter amino acid sequences, e.g., amino acid sequences of 20 or fewer amino acids, in some embodiments, substantial identity exists when one or two amino acid residues are conservatively substituted according to conservative substitutions as defined herein.

[0044] Furthermore, while exemplary embodiments are described herein, the scope of any and all embodiments includes equivalent elements, modifications, omissions, combinations (e.g., of aspects across various embodiments), applications, and / or alterations that will be understood by those skilled in the art based on this disclosure. Any limitations in the claims should be interpreted broadly based on the language used in the claims and not limited to the examples described herein or during prosecution of the application; such examples should be construed as non-exclusive. Furthermore, the acts in the processes and methods of the present disclosure can be modified in any manner, including by changing the order of acts and / or inserting additional acts and / or deleting acts. Accordingly, it is intended that the specification and examples be considered exemplary only, with the true scope and spirit being indicated by the claims and their full scope of equivalents.

[0045] The terms "individual," "subject," "host," or "patient," as used herein, have their ordinary meanings as understood by those of skill in the art, and thus include human or non-human mammals. The term "mammal" is used in its ordinary biological sense. Thus, the term specifically includes, but is not limited to, primates, including primates (chimpanzees, apes, monkeys), humans, cows, horses, sheep, goats, pigs, rabbits, dogs, cats, rodents, rats, mice, or guinea pigs. A subject according to embodiments herein may be a fetus, a newborn, a juvenile, or an adult. A "fetus" has its plain and ordinary meaning when read in light of the present specification and may include, for example, but is not limited to, a prenatal human from its embryonic state through its birth. A "neonatal" refers to a newborn child. A "juvenile" refers to an individual under the defined age of 18. An "adult" refers to an individual of any age over 18.

[0046] "Lookup table" has its plain and ordinary meaning when read in light of the present specification and may include, but is not limited to, a table made up of lists or combinations of protein variants that may exhibit increased or decreased levels of activity and / or stability, for example. The lookup table data also provides different predictive thresholds for determining the function of different amino acid substitution mutations, as different thresholds exist for different disease allele combinations.

[0047] "Cell growth," also known as "growth value," is an experimentally measured quantity that indicates the extent to which yeast cells can grow under specific environmental conditions. In some embodiments, cell growth, or "growth value," is measured by the optical density of a liquid culture. In some embodiments, cell growth, or "growth value," is measured by the area of ​​a "patch" or "spot" of cells growing on solid medium, the pixel count of a "patch" or "spot" of cells growing on solid medium, the pixel intensity of a "patch" or "spot" of cells growing on solid medium, the change in any of these parameters over time, or any combination of the foregoing parameters.

[0048] As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" means that the substance is not biologically or otherwise undesirable, e.g., the substance can be incorporated into a pharmaceutical composition administered to a subject without causing any significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is included. A pharmaceutically acceptable carrier or excipient preferably meets the required standards of toxicity and manufacturing testing and / or is listed in the Inactive Ingredients Guide prepared by the U.S. Food and Drug Administration.

[0049] "Codon optimization" or "codon optimized" refers to changes made to a nucleotide sequence such that it is more likely to be expressed at relatively high levels and translated more efficiently compared to a non-codon-optimized sequence. Codon optimization does not change the amino acids that the codons encode.

[0050] "Codon harmonization" or "codon harmonized" refers to changes made to a nucleotide sequence that do not change the identity of the amino acid coded for by the codon. Codon harmonization preserves the codon preference of the original gene and attempts to match the codon frequency at amino acid positions between two orthologous genes.

[0051] As used herein, "treatment" or "therapy" of a disease or condition refers to reducing the severity, frequency, or occurrence of at least one symptom of the disease or condition compared to a similar but untreated patient. Treatment can also refer to reducing, halting, slowing, or reversing the progression of the disease or condition compared to a similar but untreated patient. Treatment may further include addressing the underlying cause of the disease and / or one or more symptoms. The term "preventing" does not require absolute inhibition of the disorder or disease.

[0052] The terms "effective amount," "effective amount," or "effective dose," as used herein, have their ordinary meanings as understood by those skilled in the art and refer to the amount of a described composition or compound that produces an observable biological effect. Actual dosage levels of the active ingredients in the active compositions of the presently disclosed subject matter may be varied so that an effective amount of the active composition or compound is administered to achieve the desired response for a particular subject and / or application. The selected dosage level will depend on various factors, including, but not limited to, the activity of the composition, formulation, route of administration, combination with other drugs or treatments, the severity of the condition being treated, and the physical condition and past medical history of the subject being treated. In some embodiments, a minimum dose is administered, and in the absence of dose-limiting toxicity, the dose is increased to the minimum effective amount. Determination and adjustment of effective doses, as well as evaluation of when and how to make such adjustments, are contemplated herein. In the case of PSAT1 deficiency, an effective amount of a protein or composition comprising the protein disclosed herein can increase the activity of a defective protein involved in PSAT1 deficiency. In some embodiments, a therapeutically effective amount of a protein or composition comprising the protein disclosed herein can increase the stability of a defective protein involved in PSAT1 deficiency. In some embodiments, a therapeutically effective amount is an amount that prolongs the survival of a subject. In some embodiments, a therapeutically effective amount is an amount that reduces one or more symptoms associated with PSAT1 deficiency in a subject.

[0053] As used herein, an "amino acid substitution mutation" refers to any mutation understood by one of skill in the art that results in a different amino acid being encoded at that position. In some embodiments, the mutation may be a point mutation (i.e., a single nucleotide polymorphism, "SNP"). In some embodiments, the mutation may be a mutation of one, two, or all three of the nucleotides that determine which amino acid is encoded at that position in the protein.

[0054] A "conservative substitution" involves replacing an amino acid with a different amino acid with similar properties. For example, an aliphatic residue may be replaced with another aliphatic residue, a non-polar residue with another non-polar residue, an acidic residue with another acidic residue, a basic residue with another basic residue, a polar residue with another polar residue, or an aromatic residue with another aromatic residue. A "non-conservative substitution" involves replacing an amino acid with a different amino acid with different properties. Amino acids can be divided into different classes according to common side chain properties: a. hydrophobic: norleucine, Met, Ala, Val, Leu, Ile; b. neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; c. acidic: Asp, Glu; d. basic: His, Lys, Arg; e. residues that affect chain orientation: Gly, Pro; aromatic: Trp, Tyr, Phe. Non-conservative substitutions will involve exchanging a member of one such class for another class.

[0055] In some embodiments, a protein having any one of SEQ ID NOs: 1408-1537 is provided.

[0056] In some embodiments, the WT sequence is at least 80% identical to SEQ ID NO: 1 (WT sequence) and is selected from the following amino acid substitution mutations listed in Table 2: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q, K94T, P24 3A, L239M, K259T, M42I, L221I, K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A146T, A15 T, L325V, I37T, Y240N, I73M, L29V, K231N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G, I53F, T350P, P144T, G334A, R97M, R5S, N236K, P4T, T242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C, L17 Q, E229A, L63V, P4R, I37N, C80G, L357M, A310S, N55T, P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121T, S 46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, V157M, P144R, L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F. In some embodiments, the protein has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer between 80 and 100% identity to SEQ ID NO:1 (the wild-type sequence).In some embodiments, the protein has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer between 70% and 100% identity to SEQ ID NO: 1. In some embodiments, the protein has at least 80% identity to SEQ ID NO: 1. In some embodiments, the protein has at least 90% identity to SEQ ID NO: 1.

[0057] In some embodiments, the amino acid positions of SEQ ID NO: 1 are based on numbering the first amino acid at the amino terminus of the protein as position 1 and counting monotonically to the carboxy-terminal amino acid. In some embodiments, the first amino acid position is 1.

[0058] In some embodiments, protein variants of SEQ ID NOs: 1408-1537 may differ from the wild-type sequence of Figure 1 and SEQ ID NO: 1 by the substitution of one amino acid, 2, 3, 4, 5-10, 10-20, or 20-30, or 30-40, 40-50, 50-60, 60-70, or 70-80 amino acids. In some embodiments, a variant sequence may comprise the wild-type sequence of Figure 1 and SEQ ID NO: 1 in which one, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more residues have been substituted. For example, 5, 10, 15, up to 20, up to 30, or up to 40, up to 50, up to 60, up to 70, or up to 80 residues may have been substituted (using substitutions specified herein, particularly substitutions that increase the activity of PSAT1). In some embodiments, variant sequences may have one, two, three, four, five, six, seven, eight, nine, ten, twenty, thirty, or more residues substituted with conservative amino acids. In other embodiments, variant sequences may have one, two, three, four, five, six, seven, eight, nine, ten, twenty, thirty, or more residues substituted with non-conservative amino acids. In some embodiments, any one or more of the mutations listed in any one of Table 2 (or those listed in Table 1 that exhibit a ratio of activity greater than 1.05) may be combined, and may include 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, or more of the amino acid substitution mutations listed in these tables.

[0059] In some embodiments, the variants provided herein are variants that confer increased activity and / or stability to the protein. In some embodiments, the variants provided herein can confer enhanced function, stability, or activity to the protein. In some embodiments, the variants provided herein are rescue variants. In some embodiments, the variants provided herein can rescue altered function, stability, or activity of the protein. In other embodiments, the mutation has the opposite effect.

[0060] In some embodiments, the mutants specified herein can be used to enhance or rescue the function, stability, or activity of a protein when the ratio of activity of the protein variant to the wild-type protein is greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, 1.25, or higher) (e.g., those in Table 2).

[0061] In some embodiments, the mutants designated herein can be used to enhance or rescue the function, stability, or activity of a protein when the ratio of activity of the protein variant to the wild-type protein is greater than 1.05.

[0062] In some embodiments, proteins having at least one of the amino acid substitution mutations listed in Table 2 exhibit higher activity compared to the WT protein of SEQ ID NO: 1. In some embodiments, proteins having at least one of the amino acid substitution mutations in Table 1 exhibit higher activity compared to the WT protein of SEQ ID NO: 1. In some embodiments, the activity of a protein may include, but is not limited to, enzymatic activity, expression level, catalytic activity, and / or binding activity. In some embodiments, at least one or more of the amino acid substitution mutations result in a change in the activity of the protein compared to the wild-type protein. In some embodiments, the activity is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or more, or any integer between 1 and 400%, higher than the activity of the wild-type protein. In some embodiments, the ratio of the activity of a protein variant to the wild-type protein is 1.0 or higher. In some embodiments, the ratio of activity of the protein variant to the wild-type protein is greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, 1.25, or higher).

[0063] In some embodiments, the function of a protein having at least one of the amino acid substitution mutations listed in Table 2 is enhanced by more than 5% compared to the function of the wild-type protein of SEQ ID NO:1.

[0064] In some embodiments, the function of a protein having at least one of the amino acid substitution mutations listed in Table 2 is enhanced by at least 10, 20, 30, 40, 50, 100%, or more compared to the function of the wild-type protein of SEQ ID NO:1.

[0065] In some embodiments, the functionality is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 250%, 300%, 400%, or any integer between 1 and 400% greater than that of the wild-type protein. In some embodiments, the ratio of functionality of the protein variant to the wild-type protein is 1.0 or greater. In some embodiments, the ratio of functionality of the protein variant to the wild-type protein is greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, or greater), as listed in Table 2.

[0066] In some embodiments, at least one or more of the amino acid substitution mutations listed in Table 2 result in an alteration in protein expression compared to the wild-type protein. In some embodiments, functionality is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or any integer between 1 and 400% greater than expression of the wild-type protein. In some embodiments, the ratio of expression of the protein variant to the wild-type protein is 1 or greater. In some embodiments, the ratio of expression of the protein variant to the wild-type protein is 1, 1.05, 1.1, 1.15, 1.2, 1.25, or greater.

[0067] In some embodiments, at least one or more of the amino acid substitution mutations listed in Table 2 result in a change in protein abundance compared to the wild-type protein. In some embodiments, the abundance is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or any integer between 1 and 400% greater than the abundance of the wild-type protein. In some embodiments, the ratio of abundance of the protein variant to the wild-type protein is 1.0 or greater. In some embodiments, the ratio of abundance of the protein variant to the wild-type protein is greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, 1.25, or greater). In some embodiments, this is measured as the mass of the protein variant relative to the wild-type protein.

[0068] In some embodiments, at least one or more of the amino acid substitution mutations result in a change in the specificity of the protein compared to the wild-type protein. In some embodiments, the specificity is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or any integer between 1 and 400% higher than the specificity of the wild-type protein. In some embodiments, the specificity ratio of the protein variant to the wild-type protein is 1.0 or higher. In some embodiments, the specificity ratio of the protein variant to the wild-type protein is higher than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, 1.25, or higher).

[0069] In some embodiments, at least one or more of the amino acid substitution mutations result in a change in the affinity of the protein compared to the wild-type protein. In some embodiments, the abundance is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or any integer between 1 and 400% greater than the affinity of the wild-type protein. In some embodiments, the ratio of the affinity of the protein variant to the wild-type protein is 1.0 or greater. In some embodiments, the ratio of the affinity of the protein variant to the wild-type protein is greater than 1.05 (e.g., 1.051, 1.06, 1.1, 1.15, 1.2, 1.25, 1.3, 1.35, 1.4, 1.45, 1.5, 1.55, 1.6, 1.65, 1.7, 1.75, 2, 2.5, 3, 3.5, or 4 or higher).

[0070] In some embodiments, the amino acid substitution mutation results in a change in functionality, abundance, activity, specificity, affinity, or a combination of the aforementioned parameters.

[0071] In some embodiments, the sequence comprises at least one amino acid change at amino acid position 1 of SEQ ID NO: 1, wherein the amino acid positions are 2, 3, 4, 5, 7, 8, 12, 15, 17, 22, 24, 26, 29, 31, 37, 42, 46, 48, 49, 53, 55, 56, 63, 68, 71, 72, 73, 74, 80, 85, 93, 94, 95, 97, 106, 109, 110, 116, 117, 121, 125, 132, 133, 136, 138, 144, 146, 148, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 2 , 157, 169, 174, 180, 193, 198, 203, 219, 221, 226, 229, 231, 236, 239, 240, 242, 243, 245, 249, 250, 253, 255, 259, 284, 300, 302, 310, 311, 318, 325, 334, 336, 341, 344, 349, 350, 351, 353, 354, 356, 357, 359, 369.

[0072] In some embodiments, an amino acid may be substituted at at least one position designated X in Figure 2. In some embodiments, an amino acid may be substituted at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more positions designated X. For example, an amino acid may be substituted at 5, 10, 15, up to 20, up to 30, or up to 40, up to 50, up to 60, up to 70, or up to 80 positions designated X. In some embodiments, the positions indicated by X in FIG. 2 (2, 3, 4, 5, 7, 8, 12, 15, 17, 22, 24, 26, 29, 31, 37, 42, 46, 48, 49, 53, 55, 56, 63, 68, 71, 72, 73, 74, 80, 85, 93, 94, 95, 97, 106, 109, 110, 116, 117, 121, 125, 132, 133, 136, 138, 144, 146, 148, 157, 169, 174, 180, 193, 198, 203, 219, 221, 226, 229, 231, 236, 239, 240, 242, 243, 245, 249, 250, 253, 255, 259, 284, 300, 302, 310, 311, 318, 325, 334, 336, 341, 344, 349, 350, 351, 353, 354, 356, 357, 359, 369) may be substituted by conservative substitutions. In some embodiments, the amino acids at the positions indicated by X in Figure 2 (2, 3, 4, 5, 7, 8, 12, 15, 17, 22, 24, 26, 29, 31, 37, 42, 46, 48, 49, 53, 55, 56, 63, 68, 71, 72, 73, 74, 80, 85, 93, 94, 95, 97, 106, 109, 110, 116, 117, 121, 125, 132, 133, 136, 138, 144, 146, 148, 157, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 226, 229 , 174, 180, 193, 198, 203, 219, 221, 226, 229, 231, 236, 239, 240, 242, 243, 245, 249, 250, 253, 255, 259, 284, 300, 302, 310, 311, 318, 325, 334, 336, 341, 344, 349, 350, 351, 353, 354, 356, 357, 359, 369) may be replaced by non-conservative substitutions.In some embodiments, the amino acids at the positions indicated by X in Figure 2 (2, 3, 4, 5, 7, 8, 12, 15, 17, 22, 24, 26, 29, 31, 37, 42, 46, 48, 49, 53, 55, 56, 63, 68, 71, 72, 73, 74, 80, 85, 93, 94, 95, 97, 106, 109, 110, 116, 117, 121, 125, 132, 133, 136, 138, 144, 146, 148, 157, 169, 174, 175, 180, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 226, 229, 31, 37, 42, 46, 48, 49, 53, 55, 56, 63, 68, 80, 193, 198, 203, 219, 221, 226, 229, 231, 236, 239, 240, 242, 243, 245, 249, 250, 253, 255, 259, 284, 300, 302, 310, 311, 318, 325, 334, 336, 341, 344, 349, 350, 351, 353, 354, 356, 357, 359, 3690) may be substituted with a combination of conservative and non-conservative substitutions.

[0073] In some embodiments, an amino acid may be substituted at at least one position designated X in Figure 3. In some embodiments, an amino acid may be substituted at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more positions designated X. For example, an amino acid may be substituted at 5, 10, 15, up to 20, up to 30, or up to 40, up to 50, up to 60, up to 70, or up to 80 positions designated X. In some embodiments, the amino acids at the positions indicated by X in Figure 3 (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126 3, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123 , 124, 125, 126, 127, 128, 129, 130, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175 , 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 227,228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370) may be replaced by conservative substitutions. In some embodiments, the amino acids at the positions indicated by X in Figure 3 (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121 4, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157 , 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188 , 189, 190, 191, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220 , 221, 222, 223, 224, 225, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252 , 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 30 3, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 3 3, 4, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370) may be replaced with a non-conservative substitution.The amino acids at the positions indicated by X in Figure 3 (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 12 3, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119 , 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 1 84, 185, 186, 187, 188, 189, 190, 191, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216 6, 217, 218, 219, 220, 221, 222, 223, 224, 225, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248 ,249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370) may be substituted with a combination of conservative and non-conservative substitutions.

[0074] In some embodiments, an expression vector is provided comprising any one of the polynucleotide sequences of the embodiments of the present application. In some embodiments, the polynucleotide sequence is a human sequence. In other embodiments, the polynucleotide sequence is a yeast-optimized version that encodes the human sequence.

[0075] In some embodiments, an expression vector is provided that includes a polynucleotide sequence encoding one or more of the proteins of SEQ ID NOs: 1408-1537 (eg, any with increased activity).

[0076] In some embodiments, the expression vector comprises a codon-optimized sequence. In other embodiments, the expression vector comprises a sequence that is not codon-optimized.

[0077] In some embodiments, the sequence shown in GenBank Accession No. MN654100, SEQ ID NO: 1538, and FIG. 6, 1-6097 = cloning vector pAS19_yPSAT1, 1-1336 = pUC19 sequence, 1337-1577 = sequence from S. cerevisiae 3' to the SER1 (YOR184W) terminator, 1578-1839 = A. gossypii (A. An expression vector is provided that includes a terminator sequence from A. gossypii TEF, 1840-2649 = KanR aminoglycoside 3'-phosphotransferase, 2650-3083 = promoter sequence from A. gossypii TEF, 3084-3546 = terminator sequence from S. cerevisiae 3' to the SER1 (YOR184W) stop codon, 3547-4659 = yPSAT1 from Homo sapiens phosphoserine aminotransferase (codon-optimized for expression in S. cerevisiae), 4660-4755 = from S. cerevisiae 5' to the SER1 (YOR184W) start codon, and 4756-6097 = pUC19 sequence. In some embodiments, any expression vector capable of introducing genes into the yeast or human genome according to the embodiments provided herein can be used.

[0078] In some embodiments, there is provided an expression vector comprising a polynucleotide sequence encoding one or more of the proteins of any of the embodiments comprising one or more of the amino acid substitution mutations listed in Table 2. The function of the protein variants encoded by the expression vector is enhanced by more than 5% compared to the function of the wild-type protein of SEQ ID NO:1.

[0079] In some embodiments, the functionality is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 250%, 300%, 400%, or any integer between 1 and 400% greater than that of the wild-type protein. In some embodiments, the ratio of functionality of the protein variant to the wild-type protein is 1.0 or greater. In some embodiments, the ratio of functionality of the protein variant to the wild-type protein is greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, or greater), as listed in Table 2. In some embodiments, the ratio of functionality of the protein variant to the wild-type protein is greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, or higher) as listed in Table 1 for protein variants with growth estimates greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, or higher).

[0080] In some embodiments, a composition is provided comprising a protein of any one of SEQ ID NOs: 1408-1537.

[0081] In some embodiments, a composition is provided that includes an expression vector comprising a polynucleotide sequence encoding one or more of the proteins of SEQ ID NOs: 1408-1537.

[0082] In some embodiments, a composition is provided comprising a protein as described above and a pharmaceutically acceptable carrier. In some embodiments, a composition is provided comprising an expression vector encoding a protein according to any of the above-described embodiments and a pharmaceutically acceptable carrier.

[0083] In some embodiments, the protein is phosphoserine aminotransferase (PSAT1).

[0084] In some embodiments, the composition is for use in treating a genetic disorder.

[0085] In some embodiments, compositions are provided comprising a protein variant comprising one or more amino acid substitution mutations listed in Table 2. In some embodiments, the functionality ratio of the protein variant used to treat a genetic disorder relative to the wild-type protein is greater than 1.05. For example, protein variants having a functionality ratio greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, or higher) as listed in Table 2 (or those in Table 1 having a functionality ratio greater than 1.05) can be used to treat a genetic disorder. In some embodiments, two or more protein variants having the same functionality ratio can be used to treat a genetic disorder. In other embodiments, two or more protein variants having different functionality ratios can be used to treat a genetic disorder.

[0086] In some embodiments, a composition comprising at least one protein variant with enhanced functionality is administered to treat a genetic disorder. In some embodiments, the composition is administered once to treat a genetic disorder. In some embodiments, the composition is administered more than once to treat a genetic disorder. In some embodiments, the composition used in the initial treatment and the composition used in the subsequent treatments comprise the same protein variant. In some embodiments, the composition used in the initial treatment and the composition used in the subsequent treatments comprise different protein variants. In some embodiments, the composition used in the initial treatment and the composition used in the subsequent treatments comprise a combination of the same protein variant and different protein variants.

[0087] In some embodiments, compositions comprising the protein variants described herein or expression vectors encoding protein variants can be used in combination with other known agents and therapies. "Administered in combination," as used herein, means that two (or more) different treatments are delivered to a subject during the course of the subject's illness. For example, two or more treatments are delivered after the subject is diagnosed with a disorder, but before the disorder is cured or eliminated, or before treatment is discontinued for other reasons. In some embodiments, overlap exists in terms of administration, as the delivery of one treatment is still occurring when the delivery of a second treatment begins. This is sometimes referred to herein as "simultaneous" or "co-delivery." In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments, in either case, the treatments are more effective due to combined administration. For example, the second treatment is more effective, e.g., a comparable effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent than would be seen if the second treatment were administered in the absence of the first treatment, or a similar situation is seen with the first treatment. In some embodiments, delivery is performed such that the reduction in symptoms or other parameters associated with the disorder is greater than that which would be observed with one treatment delivered in the absence of the other treatment. The effect of the two treatments may be partially additive, wholly additive, or greater than additive. Delivery may be performed such that the effect of the first treatment administered is still detectable upon delivery of the second treatment.

[0088] In some embodiments, the first treatment comprises administering to the subject a protein comprising a protein variant or a combination of amino acid substitution mutations. In one aspect, the subsequent treatment comprises administering to the subject the same protein variant or a protein comprising the same combination of amino acid substitution mutations as the first treatment. In other embodiments, the subsequent treatment comprises administering to the subject a protein variant that is different from the first treatment or a protein comprising a different combination of amino acid substitution mutations.

[0089] In some embodiments, a composition comprising a protein variant is administered to a subject with a genetic disorder in an amount effective to correct the genetic disorder or a symptom thereof in the subject.

[0090] In some embodiments, one or more expression vectors encoding a protein variant according to any of the embodiments are administered to a subject with a genetic disorder in an amount effective to correct the genetic disorder or a symptom thereof in the subject.

[0091] In some embodiments, the variant sequence administered to a subject with a genetic disorder differs from the wild-type (WT) sequence of SEQ ID NO: 1 by one amino acid, 2, 3, 4, 5-10, 10-20, 20-30, or 30-40, 40-50, 50-60, 60-70, or 70-80 amino acid substitutions. In some embodiments, the variant sequence may comprise the wild-type sequence of FIG. 1 and SEQ ID NO: 1 in which one residue, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more residues have been substituted. For example, 5, 10, 15, up to 20, up to 30, or up to 40, up to 50, up to 60, up to 70, or up to 80 residues may have been substituted. In some embodiments, a single variant sequence containing up to 5, 10, 15, 20, 30, or more amino acid substitutions may be administered. In other embodiments, combinations of variant sequences of SEQ ID NOS: 1408-1537 may be administered. In some embodiments, any one or more of the mutations listed in Table 2 may be combined, and may include 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, or more of the amino acid substitution mutations listed in such tables. In some embodiments, any one or more of the loss-of-function mutants in Table 3 may be combined.

[0092] In some embodiments, the subject is a mammal such as a rodent (e.g., guinea pig, hamster, rat, mouse), murine (e.g., mouse), canine (e.g., dog), feline (e.g., cat), equine (e.g., horse), primate, simian (e.g., monkey or ape), monkey (e.g., marmoset, baboon), ape (e.g., gorilla, chimpanzee, orangutan, gibbon), or human. In some embodiments, the subject is a human. In other embodiments, a non-human mammal is used, particularly a mammal traditionally used as a model for demonstrating therapeutic efficacy in humans (e.g., murine, primate, pig, canine, or lagomorph). In some embodiments, the subject is a fetus, newborn, juvenile, or adult. In some embodiments, the subject has a complete deficiency of the PSAT1 enzyme. The PSAT1 enzyme is a pyridoxal phosphate (PLP)-dependent homodimer and is the second enzyme in the serine biosynthetic pathway. The PSAT1 enzyme is highly conserved from bacteria to humans, and the human cDNA sequence can functionally replace the orthologous gene SER1 in Saccharomyces cerevisiae. In some embodiments, the subject has a partial absence of PSAT1. In some embodiments, the subject is suspected of having PSAT1 deficiency (PSATD) (MIM:610992), which is manifested by a deleterious mutation in the human PSAT1 gene. The subject experiences symptoms consistent with a diagnosis of PSATD. The subject does not experience any symptoms or does not exhibit symptoms inconsistent with PSATD. The subject can be diagnosed with PSATD based on DNA sequencing-based methods. The subject can be diagnosed with PSATD based on biochemical assays. The subject does not have PSATD that is detectable by DNA sequencing-based methods and / or biochemical assays. In some embodiments, the subject is at a higher risk of having PSATD. In some embodiments, the subject has a family history of PSATD. In some embodiments, the subject has a genetic predisposition to PSATD. In some embodiments, the subject is not at higher risk of having PSATD.

[0093] According to embodiments described herein, compositions comprising protein variants can be administered by any suitable route of administration. Without limitation, compositions comprising protein variants can be administered to a subject orally, rectally, transdermally, intramuscularly, intravenously, intranasally, or by inhalation. In some embodiments, compositions comprising protein variants are administered to liver cells. In other embodiments, compositions comprising protein variants are administered to the blood, spleen, or lungs. In some embodiments, compositions comprising protein variants can be administered by injection or in the form of a tablet, capsule, or drink. In some embodiments, compositions comprising protein variants of the present invention can also be used for gene therapy using standard gene delivery protocols. Methods for gene delivery are known in the art. See, e.g., U.S. Pat. Nos. 5,399,346 and 5,580,859. These patents are incorporated herein by reference in their entireties. In some embodiments, gene therapy is mediated by a gene editing enzyme system. Some non-limiting examples of gene editing enzyme systems include CRISPR / Cas9 systems, zinc finger nuclease systems, transcription activator-like effector nuclease (TALEN) systems, homing endonuclease (HE) systems, or meganuclease systems.

[0094] As described herein, administration of a composition comprising a protein variant can modify, treat, inhibit, or alleviate PSATD (MIM:610992), Neuraxova syndrome 2 (NLS2) (MIM:616038), or symptoms associated therewith. A few non-limiting examples of symptoms associated with PSATD are congenital microcephaly, seizures, psychomotor retardation, and spastic quadriplegia. A few non-limiting examples of symptoms associated with NLS2 (the severe end of the PSATD case spectrum) are characteristic facial features with shortened eyelids, microcephaly, intrauterine growth retardation (IUGR), skin abnormalities (e.g., ichthyosis and hyperkeratosis), flexion deformities, limb malformations, edema of the hands and feet, abnormal rotational movements, corpus callosum hypoplasia, or neural tube defects. Severe malformations manifested by NLS2 can lead to prenatal or early postnatal death. As disclosed herein, "correction" is used broadly to refer to the replacement or suppression of a protein variant associated with a PSATD, including NLS2. In some embodiments, the replacement of a protein variant associated with a PSATD is achieved by gene therapy. In some embodiments, the replacement of a protein variant associated with NLS2 is achieved by gene therapy. In some embodiments, the protein variant is replaced with a wild-type protein. In other embodiments, the protein variant is replaced with a high-functioning protein variant. As disclosed herein, palliative is used broadly to refer to at least a reduction in a parameter associated with the pathological condition being treated, e.g., the magnitude of a symptom. In some embodiments, the method can completely inhibit, e.g., prevent the onset, or arrest, e.g., terminate, such that the host no longer suffers from at least one or more of the pathological condition or symptoms characterizing the pathological condition. In some embodiments, the method can delay or slow disease progression, whether detectable or undetectable, or can alleviate or relieve, or ameliorate (whether partial or complete), the disease state.

[0095] When an "effective amount" is indicated, the exact amount of the composition of the present invention to be administered can be determined by a physician taking into account individual differences in the age, sex, weight, severity of symptoms, progression of the disease, and condition of the subject.

[0096] In some embodiments, a method is provided for enhancing the functionality of PSAT1 protein in a subject, the method comprising administering a protein or composition to the subject, wherein after administration, PSATD, NLS2, or symptoms thereof are reduced.

[0097] In some embodiments, enhancing the functionality of PSAT1 can enhance the abundance, activity, specificity, binding affinity, or a combination of the aforementioned parameters of PSAT1. In some embodiments, the activity of the protein may include, but is not limited to, enzymatic activity, non-enzymatic activity, catalytic activity, non-catalytic activity, and binding activity. In some embodiments, the functionality of the PSAT1 protein in the subject before administration is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% lower than the functionality of the wild-type protein. In some embodiments, the ratio of functionality of the wild-type protein to the protein variant before administration is lower than 0.99 (e.g., 0.98, 0.95, 0.94, 0.93, 0.92, 0.91, or lower). In some embodiments, the functionality of the PSAT1 protein after administration is increased by at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 250%, 300%, 400%, or any integer between 1 and 400% compared to the functionality of the wild-type protein. In some embodiments, the ratio of functionality of the wild-type protein to the protein variant after administration is higher than 1.05 (e.g., 1.06, 1.10, 1.15, 1.20, or higher).

[0098] In some embodiments, the subject is a fetus, a neonate, a juvenile, or an adult.

[0099] In some embodiments, methods are provided in which one or more expression vectors are administered to a subject in an amount that corrects the PSATD or a symptom thereof in the subject.

[0100] In some embodiments, PSAT1 is modified to exhibit enhanced function.

[0101] In some embodiments, PSAT1 is replaced or complemented with one or more polynucleotide sequences encoding SEQ ID NOs: 4-1537 to exhibit enhanced function.

[0102] In some embodiments, the function of a protein having at least one of the amino acid substitution mutations listed in Table 2 is enhanced by at least 5% compared to the function of the WT protein of SEQ ID NO:1.

[0103] In some embodiments, the function of a protein having at least one of the amino acid substitution mutations listed in Table 2 is enhanced by at least 50% compared to the function of the WT protein of SEQ ID NO:1.

[0104] In some embodiments, the function of the PSAT1 protein is enhanced by at least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or any value between 5% and 400%. In some embodiments, the functionality of the PSAT1 protein in the subject prior to administration is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% lower than the functionality of the wild-type protein. In some embodiments, the ratio of functionality of the protein variant to the wild-type protein before administration is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower). In some embodiments, the functionality of the PSAT1 protein after administration is increased by at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 250%, 300%, 400%, or any integer between 1 and 400% over the functionality of the wild-type protein. In some embodiments, the ratio of functionality of the protein variant to the wild-type protein after administration is greater than 1.05 (e.g., 1.06, 1.10, 1.15, 1.20, or higher). In some embodiments, subjects with mutant protein levels lower than wild type (e.g., any value in the tables herein at or below 1.0) can be cured by adding mutant protein levels higher than wild type (e.g., any value in the tables herein above 1.05).

[0105] In some embodiments, PSAT1 is replaced or complemented with one or more polynucleotide sequences encoding SEQ ID NOs: 1408-1537 to exhibit enhanced stability.

[0106] In some embodiments, the stability of the PSAT1 protein is enhanced by at least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or any value between 5% and 400%. In some embodiments, the stability of the PSAT1 protein in the subject prior to administration is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or any integer between 1 and 95% less than the stability of the wild-type protein. In some embodiments, the stability ratio of the protein variant to the wild-type protein before administration is 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower. In some embodiments, the stability of the PSAT1 protein after administration is increased by at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 250%, 300%, 400%, or any integer between 1 and 400% over the stability of the wild-type protein. In some embodiments, the stability ratio of the protein variant to the wild-type protein after administration is greater than 1.05 (e.g., 1.06, 1.1, 1.15, 1.2, or higher).

[0107] In some embodiments, a kit for determining a specific treatment mode for a genetic disorder is provided. The kit includes a reference table indicating specific amino acid substitutions commonly involved in cellular mechanisms, and one or more vectors are provided that include the sequences of SEQ ID NOs: 4-1537.

[0108] In some embodiments, a lookup table is provided that includes a list of the mutations listed in Table 2 and Table 3. In some embodiments, the table includes mutations that confer reduced activity and / or stability to the protein (Table 3, or ratios less than 0.99 in Table 1). In some embodiments, the table includes mutations that confer increased activity and / or stability to the protein (Table 2, or ratios greater than 1.05 in Table 1).

[0109] In some embodiments, the reference table lists a panel containing multiple protein variants and their effects on protein functionality. In some embodiments, reference to the reference table can be used to select an amino acid substitution that reduces, alleviates, or limits symptoms associated with a genetic disorder. In other embodiments, when it is not possible to revert a pathogenic amino acid substitution mutation to the reference amino acid, reference to the reference table can provide functional information for selecting an alternative functional amino acid substitution. Regions of a protein can be selected in which the frequency of amino acid substitutions is significantly higher in subjects with a genetic disorder than in a reference population without the disorder. In some embodiments, reference to the reference table can be used to narrow down regions of a protein for gene therapy.

[0110] In some embodiments, the genetic disorder is PSATD.

[0111] In some embodiments, a method for identifying a subject with a genetic disorder is provided, comprising detecting the presence of at least one mutation in a protein that differs in a subject with the genetic disorder compared to a subject without the disorder, wherein the presence of the at least one mutation indicates the presence of the genetic disorder in the subject. These mutations may be those listed in Table 3 and / or SEQ ID NOs: 4-1407.

[0112] In some embodiments, the presence of at least one mutation can be used to indicate the presence of a genetic disorder in a high-risk patient population earlier than is possible with existing methods for detecting genetic disorders. In some embodiments, the presence of at least one mutation can be used to indicate the presence of a genetic disorder in a subject who is not at risk of developing the genetic disorder. In some embodiments, the presence of at least one mutation in a subject indicates that the subject has a genetic predisposition to developing the genetic disorder. In some embodiments, the presence of at least one mutation in a subject indicates that the subject is in an early stage of the development of the genetic disorder. In some embodiments, the presence of at least one mutation in a subject indicates that the subject is in an advanced stage of the genetic disorder.

[0113] In some embodiments, the ratio of functionality of the wild-type protein to the protein variant with at least one mutation detected in the subject is less than 1.0. In some embodiments, the ratio of functionality of the protein variant with at least one mutation detected in the subject to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower), as listed in Table 3. In some embodiments, the ratio of functionality of a protein variant having at least one mutation detected in a subject to a wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower) as listed in Table 1 for those with a growth estimate of less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower).

[0114] In some embodiments, one or more regions of the protein may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, or more amino acid substitutions. Many of these amino acid substitutions may be clinically actionable, and the observed abnormality may be indicative of a genetic disorder. In some embodiments, the abnormality may be a decrease in protein function compared to a reference subject without the disorder. In some embodiments, the abnormality may be a decrease in protein stability compared to a reference subject without the disorder.

[0115] In some embodiments, a method for ameliorating, alleviating, or treating a genetic disorder is provided. The method comprises detecting the presence of at least one mutation in a protein that differs in a subject with the genetic disorder compared to a subject without the disorder, wherein the presence of the at least one mutation indicates the presence of the genetic disorder in the subject. The method also comprises administering to the subject at least one protein variant or composition provided in the embodiments herein, wherein after administration, the genetic disorder or its symptoms are reduced.

[0116] In some embodiments, the mutation to be detected is 3 (Table 3), C245R, T242M, G206R, N89Y, G119E, G13R, L40R, I25K, G78V, L63S, G78D, Y148N, G79R, S147Y, I25R, P12L, G203R, I121K, G78A, V39D, H44Y, C152F, T156M, S179L, E155Q, S84R, G11R, D176H, G79W, A99D, G78R, C152R, G119R, D176A, G105V, K200N, Y151D, D176Y, S108P, G197D, Q82R, R 45S, G78C, A111P, V86D, N154Y, G13S, Q199P, Y148H, R342Q, G159R, K200Q , Y148D, K200T, E155V, A115D, L182R, A111D, Y101D, G79E, K200R, G197R, C 152Y, D176N, E155K, A198D, K200M, R97G, V188D, P14T, P12R, N89K, S38R, Y101N, P87R, G13C, L255P, T104P, N154D, A109P, L90P, Q82P, N302Y, P14L, P14H, Q199K, T156P, T156R, C224W, R61P, S178P, C152W, A112P, T242P, L3 57R, L90R, H335D, N302K, W107R, L345R, Y289D, S108L, Y148S, Y101S, K200 E, L255R, V174D, G78S, M251R, N201Y, S147F, W107C, G203D, T242R, V102E , S299R, Q199L, G203V, T242K, S344Y, R213P, D176E, S299I, R342W, Y289S, V174F, W107L, S147P, L332W, S344F, M361R, Y70D, R342P, R306P, I278N, Y 150N, P243R, C291F, R342L, V212D, G105A, T156A, H158P, W107S, Y148C, M3 61K, R45M, V157E, C175W, Y289N, R342G, V193E, Y346N, G340R, A99P, H44L, E155G, V202D, S38I, I194N, Y70N, S178F, V102G, S178Y, R45G, V72G, N89I,Y101C、Y289H、R45K、N241I、R45T、Y70S、Y346S、I91S、V103G、F10S、G334W、H335L、R213G、I278S、C175R、V149E、Y151N、N89D、I282N、N302H、C291R、Q199E、S179P、G340D、I307N、A115T、I91T、P243H、V72E、S183P、V149G、L332S、S285Y、Q76R、D353V、F305S、A198P、A99T、A324P、H44N、P244R、S272P、M329R、E155A、H44P、G11V、Y346C、L64P、V174G、G119V、C175F、G105E、N154I、T156S、S275P、L270P、A111V、G340C、F181S、S247R、H335P、E155D、H44Q、F191S、I303N、T104K、G119W、Q276P、P14A、L59P、N122I、L17R、A115V、P14S、G11W、S299C、L22W、F364L、R45W、N241Y、Y70C、L75R、N201H、G334V、I121R、P87H、Y230D、V124D、L216R、D187Y、P87L、S179A、G170V、Y289C、A358P、V293M、S178T、G287V、P67L、A205D、W140L、N154T、P292R、V293E、G339V、S275Y、M361T、Y150C、V60E、A343D、W140R、S204Y、F364S、V193G、F83Y、V86G、H335R、A99V、P292Q、Y151S、L321P、R213L、I211S、N89T、E161K、F320C、N180K、S147T、F10I、G263V、P292A、C245Y、I282S、P166L、F364C、F288C、G340S、P166T、I282T、P292S、I303F、E161V、I278T、A111T、G197V、A153P、P166H、C245F、R213C、L64R、M361V、H335Q、T104R、H335N、S344P、S285C、P166S、L216P、F320I、W140G、V338E、G206V、V66F、L228P、F364V、F195S、S43R、Q233P、W140C、Y279H、V188G、R61W、R298I、A146D、D283H、L63W、I303S、M42R、C224Y、S285T、V209E、F320L、F364I、V174L、L357Q、R298S、I307S、V103E、C175Y、E317D、R298T、A106T、I341N、L216Q、N180Y、E317G、T139P、V172G、D163V、A112E、S285A、A324D、E366V、I282F、H158Y、F181C、Y279D、A359P、F191V、N347Y、I281F、S275T、K274I、P12T、Y279F、I303M、G92V、P292L、P166A、R213S、V293A、M177K、V186G、R298K、D163Y、N328I、I134L、V210E、V60G、R298G、V202G、Y279S、D353A、G206E、V21E、I134T、N89S、A264D、C80R、T277P、D100Y、F49S、Q233R、F320Y、F162V、G197S、G262V、Y230S、I134N、A15D、K94I、H368R、Y70H、I134S、I307T、D353Y、A358D、E114K、V290G、V160M、V160G、F118S、Q286P、S204P、V124G、T56R、H158R、M329I、N347D、E161D、F288S、F162I、I248S、K333N、N180H、L182Q、R61L、F10V、T104I、A15P、F288V、K33I、W257S、K274E、M267K、N302D、I281N、F191L、G79A、N347T、D100G、I211T、F305L、F83C、E366K、I303T、L182P、C291S、V186E、V293L、M329V、L316S、I281S、F162S、D163H、D100V、F288I、M267R、A348G、D283V、A99G、S204F、P67R、R5G、P225L、Y230H、L17M、V124F、F162C、V102L、I134F、I278F、G77E、V21G、V160A、V209G、F288L、F360S、C224R、F181L、Y230C、N89H、S189F、I282M、F191C、D314A、L75V、F10C、S178C、S20L、T56P、P67A、R306G、N69I、I258T、I134M、K190E、E57K、S179T、P126A、F305Y、H158L、H335Y、I341L、F305V、E317K、T277R、F83I、P87S、P244L、F83V、F49C、D187G、D283G、L316V、P4L、D100A、K190N、A15V、K356R、A220D、E280K、S108T、Y32D、G263R、V354G、S84T、M361L、I307F、V250F、P225H、F74S、E268V、E114A、E366A、S299T、N284I、A234P、Y101H、D353G、V168D、G218W、P67T、A106P、C152G、P18Q、I194S、C98F、D68Y、A99S、L90F、V202A、N54I、V103M、V290L、P67Q、Y151C、F49L、N58K、Y230N、L327P、V338G、G203C、V102A、T242A、R61Q、F49V、V354A、P135L、K190T、I194F、A109V、H125D、D163A、S46P、Y101F、S337P、K51Q、L59V、N297T、V124A、I307M、A266P、E280V、G159A、C224F、V149A、K116R、N154H、G263D、I52M、Y240D、Y346H、E352V、E229D、F181V、V86F、G197A、D187V、A106G、N54T、F162L、F181I、K333Q、D136A、P67S、D136G、G339E、D68A、W257C、G262A、K274R、S84I、I248N、G159C、K71N、M42V、V349D、I282L、N328Y、L228R、Q76E、V212G、S84C、R222G、E326Q、V35F、S43I、V7G、S183T、Q76H、K190R、V66L、S130R、Q276E、F305I、L40I、G287R、T104S、A343G、D167V、G235V、A220V、V254D、G77V、V157G、R213H、L239V、S238P、D187H、N302S、S275C、G339A、L217M、A196T、A220P、T208I、D100E、S247C、A264G、E62V、E161Q、V290D、S183C、Q6H、D167Y、H19L、I165L、S299G、S20P、Y32F、C98W、C175S、A153T、I258S、E161G、P137R、N284Y、L216M、D187A、A343S、Y279N、K51R、I123F、V21A、S130T、G36C、G308D、V188F、P126R、D215V、K33R、V207F、D100H、S183Y、N328S、I91V、K127R、L217R、I91F、N143K、N201K、V21L、N297D、G262R、G312V、P135T、N69D、N55I、V60A、N69Y、V250D、I165R、K300T、N201I、K323N、I53N、S20T、Q276L、K190Q、L128V、S204C、D187N、F83L、D215N、D353N、H19Q、G119A、E57D、G334E、A265T、Y32H、I73S、S108A、V35I、G81S、V186M、I165K、S331T、L23I、E317Q、V39L、T139S、P126L、H19D、P18A、V212F、G81C、H158Q、L173P、A348V、L316F、N180I、E57V、G235E、P304A、D145V、E24D、A112T、K323R、Q296P、S43G、I351V、N69K、G334R、R222P、T120S、E28A、E366G、V168I、V72A、D353H、I341S、L93S、L142H、T139N、D31E、P137T、F181Y、N309D、V103A、L75P、A85V、H125P、R222L、D163E、D215H、K333E、G129W、N309I、Q369L、G170A、R306C、S247I、V103L、P18R、N347H、E57G、I25M、P244S、W257G、L325H、I273S、I123V、K300E、I53S、Q82L、L327H、L270R、G36V、A266V、V21M、I282V、K300M、S271G、S189P、A50G、C224S、V124L、I52L、T277A、E62Q、K323Q、D313Y、A171S、N328K、I52T、I165M、K27Q、K16R、E24G、F288Y、L255Q、A220T、F195C、T132R、Q286R、G92A、V188A、Q369R、H125L、E256Q、T56A、E268K、P137Q、N54D、L59I、E57A、S147A、K94E、K51M、S237N、V60L、L253M、N236H、G235A、I2 11F、K16Q、A65V、D283A、D163G、G287A、P225A、K127N、K33E、F364Y、L59H、V3 9F、A109G、A171V、V227D、T277I、L217Q、M177T、H19P、F164S、G263C、E223V、L216V、D48N、L217V、E229Q、P295L、E113V、F10L、F164I、I303V、S38G、K323E、 N58T、N297H、M267T、R5T、V186A、N58I、K27T、A146P、G308A、K133I、P244T、G 252V、C224G、G92S、G81V、V202L、V168L、V290I、T104A、P185A、Y240H、E28Q、 L345M、G36R、A198V、S272T、K333I、I341M、G263S、E280G、K274T、I165T、D2A、Q296H、A109D、E114V、Q355H、K318E、V254I、D322V、A153S、C98Y、D215Y、I73 N、N241T、N141H、K363N、V35L、T120P、I281V、P225S、V212I、A109S、D68G、K2 74Q、L221P、D215G、L30F、E24V、L64V、D100N、M177V、L128F、G92C、V160L、A1 98G、E28D、P87T、S147C、E268A、R5M、V290A、A85T、P225T、S247G、R319G、H19R、L365W、I307V、L142V、E223K、L128R、S183A、T132I、W257R、G312R、P135A、D 313A、Q276K、P87A、S204A、S38C、K362R、V207L、V250I、S331Y、T139A、A3D、A65D、E294G、H368P、A146G、S237I、T139I、F164Y、N54Y、K184N、R61G、V102M、 V60M、I307L、I52N、K363R、D187E、E317A、V149L、E62A、S130C、A265S、D145E 、V202I、N122H、Q82E、L64Q、M267L、L59F、S130G、I248F、D145A、K16E、S237G、D167E、Y150H、Q286L、N297Y、A95E、K300N、S247T、K33Q、I123T、K269E、M251T、A153G、Y230F、P135R、F164V、A171P、D145Y、N328T、L270V、I25V、I121L、S189C、A266S、V35G、C98S、M301T、V212L、V7M、D167N、I121M、T350I、P295T、A358V、K190M、P126S、Q276H、G287E、A3S、D215E、K127T、N58Y、F164C、Q296E、F118Y、L142I、L40V、G218A、G96V、M267V、N284D、L88I、D167H、V8D、P185L、P244A、T277S、A310V、R5K、L330M、F164L、L217P、A343P、N141S、N58H、D31V、G312E、L270F、S237C、E268G、A324S、F74V、D314Y、D2V、L370I、N55S、I25L、E366D、L345P、A265P、L17V、A315P、K333R、K363E、V254L、Q369P、V86A、T120I、V254A、D48V、Y70F、E268Q、N261T、N309T、P244Q、K269M、G92D、C152S、N141Y、S238Y、G218R、N9S、V232L、I258L、N284S、E256D、S84N、Q369H、A146S、K231E、V254G、H368D、L253W、E256V、G262E、R306S、Q233K、I278M、E294V、V174A、N261I、S43C、K71R、S138C、L325P、D322G、D2G、A358T、A106V、K269N、V232E、E114G、F360I、P295A、L253V、V210G、V124I、I123M、V207G、I278V、K318I、F246S、E113D、A3T、K27M、S46L、L327R、G218E、L88S、A65G、Q286E、Q296K、K184R、P18S、Q6R、F360Y、D48A、A146V、I273L、L332V、N141D、V168A、V232G、I53V、L59R、T277K、N328D、Q233H、S272C、I52S、L75Q、S130N、L327F、N141K、D322N、S138I、D314H、Y151H、L142R、K274N、G96A、L40F、A234S、K184E、N58S、L182V、E294K 、M301K、P12H、L327V、K362T、I91L、G81A、I73T、I351S、K362Q、L40H、L330V、 Q26L、N309Y、V212A、A266D、G308S、F360C、A111G、D68H、S247N、A50D、S331P、K363Q、E326A、K362N、K33N、E229V、Y249D、N261K、K27N、G170R、K311N、A220 S、P4A、I278L、A112V、I273N、T132P、L221F、S237T、D214H、L142P、G218V、G3 6D、I341F、E268D、L142F、G308R、A315S、D31N、A264P、R306L、S47A、G34V、I21 1V、I258N、E223Q、N284H、A50S、P295S、Q6E、A358G、L321H、D214A、A205S、V1 88I、Y240F、I273M、F118L、D313E、D314G、G36A、I273V、N122T、R319I、A65S、C 291G、G34R、F219Y、S271R、Q276R、M42L、G339R、E57Q、V168G、P18L、K169N、D322E、A310P、E113A、V66I、K259I、S331A、N260H、L29S、I123L、I52F、S337T、 M367R、L90V、K231Q、F219V、K127Q、D214G、P126H、A324T、I194T、Y249S、Q76 P、I37S、D145N、G77A、D314E、E294Q、K127E、D313G、S138N、A324G、D48H、Y240 C、D68N、G129A、I281L、V202F、L365S、D322A、E280D、I273F、K110R、K27E、E161A、N201S、F219L、V193A、L239W、K363T、K269T、I248V、N143D、D31G、Q355L、 E280Q、A196V、L93W、W257L、K323T、Q82H、V250L、S272Y、G252D、D313H、P304R、L270H、Q6P、A264S、I211M、L128I、N309K、V86I、N122D、N55Y、M367V、H19N、V172E, H368N, K184T, K16T, D136N, R97K, A266T, R336T, N260I, N260Y, D2N, S272F, I351L, M329L, L321V, A343V, G129E, N54K.

[0117] In some embodiments, the mutants provided herein are mutants that result in protein variants with reduced activity and / or stability.

[0118] In some embodiments, a mutant is classified as one that results in a protein variant with reduced activity and / or stability if the ratio of activity of the protein variant to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower).

[0119] In some embodiments, a mutant is classified as one that results in a protein variant with reduced activity and / or stability if the ratio of activity of the protein variant to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower).

[0120] In some embodiments, the functionality of a mutant resulting in a protein variant with reduced activity and / or stability is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% less than the functionality of the wild-type protein. In some embodiments, the ratio of functionality of a protein variant to the wild-type protein is less than 1.0. In some embodiments, the ratio of functionality of a protein variant with reduced activity and / or stability to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or less), as listed in Table 3.

[0121] In some embodiments, the activity of a mutant resulting in a protein variant with reduced activity and / or stability is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% less than the activity of the wild-type protein. In some embodiments, the ratio of activity of the protein variant to the wild-type protein is less than 0.99 or 1.00. In some embodiments, the ratio of activity of a protein variant with reduced activity and / or stability to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or less). In some embodiments, the activity of a protein may include, but is not limited to, an enzymatic activity, a non-enzymatic activity, a catalytic activity, a non-catalytic activity, and a binding activity.

[0122] In some embodiments, expression of a mutant resulting in a protein variant with reduced activity and / or stability is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% lower than expression of the wild-type protein. In some embodiments, the ratio of expression of the protein variant to the wild-type protein is less than 0.99 or 1.00. In some embodiments, the ratio of expression of the protein variant with reduced activity and / or stability to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower).

[0123] In some embodiments, the abundance of a mutant resulting in a protein variant with reduced activity and / or stability is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% lower than the abundance of the wild-type protein. In some embodiments, the ratio of abundance of the protein variant to the wild-type protein is less than 0.98 or 0.99. In some embodiments, the ratio of abundance of the protein variant with reduced activity and / or stability to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower).

[0124] In some embodiments, the specificity of a mutant resulting in a protein variant with reduced activity and / or stability is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% less than the specificity of the wild-type protein. In some embodiments, the specificity ratio of the protein variant to the wild-type protein is less than 0.99. In some embodiments, the specificity ratio of the protein variant with reduced activity and / or stability to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or less).

[0125] In some embodiments, the affinity of a mutant resulting in a protein variant with reduced activity and / or stability is at least 0%, 1%, 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or any integer between 1 and 99% less than the affinity of the wild-type protein. In some embodiments, the ratio of the affinity of the protein variant to the wild-type protein is less than 1.00. In some embodiments, the ratio of the affinity of the protein variant with reduced activity and / or stability to the wild-type protein is less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or less).

[0126] In some embodiments, mutations resulting in protein variants with reduced activity and / or stability exhibit altered functionality, abundance, activity, specificity, affinity, or a combination of the aforementioned parameters compared to the wild type.

[0127] In some embodiments, the presence of one or more variants resulting in a protein variant with reduced activity and / or stability in a subject can indicate the presence of a genetic disorder in the subject. In some embodiments, a composition comprising one or more variants resulting in a protein variant with increased activity and / or stability as provided in the embodiments is administered to a subject with a genetic disorder. In some embodiments, administration of one or more variants resulting in a protein variant with increased activity and / or stability can replace one or more variants resulting in a protein variant with reduced activity and / or stability in a subject. In other embodiments, administration of one or more variants resulting in a protein variant with increased activity and / or stability can suppress one or more variants resulting in a protein variant with reduced activity and / or stability in a subject. In other embodiments, administration of one or more variants resulting in a protein variant with increased activity and / or stability can enhance the functionality of the variants resulting in a protein variant with reduced activity and / or stability in a subject.

[0128] In some embodiments, the genetic disorder is PSATD.

[0129] Development of assays to determine highly functional protein variants As provided herein, assays were developed to determine the effect of various protein variants on the functionality and stability of PSAT1.

[0130] The construct encoding the protein variant can be used for expression in a yeast system in which the yeast homologue is knocked out.For example, yeast can be genetically modified to knock out the gene SER1, which is the yeast ortholog of the human PSAT1 protein.The yeast can then be genetically modified to express PSAT1 from nucleic acid.Expression constructs for yeast are known to those skilled in the art and can have a strong yeast promoter / terminator and can include a yeast selection marker cassette.

[0131] Assays can then be performed to determine yeast cell growth relative to yeast cells expressing wild-type PSAT1 or yeast cells expressing a PSAT1 gene without the mutation. Loss or enhancement of function of the yeast enzyme produces a quantitative and easily measured trait: poor growth in the absence of serine complementation.

[0132] In some embodiments, the assay is performed using the steps provided herein. In some embodiments, the assay involves: 1) growing yeast cells expressing wild-type PSAT1 or a variant protein to saturation in a rich liquid medium; 2) then plating the yeast cells onto solid medium (YPG: 1% yeast extract, 2% peptone, 2% glycerol, 2% Bacto Agar) using glycerol as the carbon source to remove respiratory-deficient yeast cells; 3) then plating the yeast cells back onto a rich liquid medium (YPD: 1% yeast extract, 2% peptone, 2% glucose) and growing to saturation; 4) then plating the yeast cells in duplicate (n=3-6) onto solid minimal medium lacking serine (SD: 1.7 g / L Bacto Yeast Nitrogen Base without amino acids or ammonium sulfate, 0.5% ammonium sulfate, 2% glucose, 2% Bacto Agar) and growing at 30°C for 3 days; and 5) plating is performed using a Biomek with a V&P 96 pinhead. The assay may include one or more of the following: performing the assay using an i7 robot; In some embodiments, the assay may be used for any relative comparison of functionality provided herein, including the claims.

[0133] In some embodiments, functional comparisons of the various protein variants disclosed herein to the WT protein are made using the assays provided herein.

[0134] kit Also provided herein are kits that include the expression constructs, protein sequences provided and described herein, and written instructions for making and using them. Thus, for example, provided herein are kits that include one or more of the protein sequences described herein and / or expression constructs described herein.

[0135] The kit also provides a lookup table for use in evaluating combinations of mutations that exhibit enhanced functionality or stability. A lookup table is a table that correlates one form of data with another form of data, or correlates one or more forms of data with a predicted outcome, such as a phenotype or trait, to which the data is associated. For example, the lookup table may include a correlation between allele data for at least one protein involved in a metabolic pathway and a particular trait or phenotype, such as a particular disease diagnosis, that individuals with the particular allele data are likely to exhibit or are more likely to exhibit than individuals without the particular allele data. The lookup table may be multidimensional, for example, but not limited to, simultaneously including information on multiple alleles of a particular protein or proteins, or including information on multiple protein variants, such as mutational variants, and may also include other factors, such as details regarding disease diagnosis, treatments, or drugs used for a particular disease. In some embodiments, the lookup table includes a correlation between at least one allele and a disease.

[0136] The look-up table can be generated by individual evaluation of protein variants or by studying cells carrying mutant forms of the protein to be tested.

[0137] Development of dietary treatments after protein variant detection Also provided herein are methods for ameliorating, alleviating, or treating a genetic disorder, the methods comprising: 1) receiving diagnostic information comprising the detection of the presence of a DNA variant resulting in at least one mutation in a protein that differs in a subject with the genetic disorder compared to a subject without the disorder, wherein the presence of the at least one mutation indicates the presence of the genetic disorder in the subject; and 2) administering a therapeutic agent to the subject, wherein the genetic disorder or its symptoms are reduced after administration. In some embodiments, the therapeutic agent comprises an effective amount of a dietary supplement. In some embodiments, the dietary supplement comprises L-serine. In some embodiments, the dietary supplement comprises glycine. In some embodiments, the dietary supplement comprises L-serine and glycine. In some embodiments, an effective amount of L-serine comprises 100 mg / kg / day, 300 mg / kg / day, 400 mg / kg / day, 500 mg / kg / day, or 950 mg / kg / day. In some embodiments, an effective amount of L-serine comprises 100 to 950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 300-950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 400-950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 500-950 mg / kg / day. In some embodiments, the effective amount of glycine comprises 200 mg / kg / day. In some embodiments, the effective amount of glycine comprises 400 mg / kg / day. In some embodiments, the dietary supplement comprises L-serine and glycine. In some embodiments, the effective amount comprises 500 mg / kg / day of L-serine and 200 mg / kg / day of glycine. In some embodiments, the effective amount comprises 300 mg / kg / day of L-serine and 400 mg / kg / day of glycine. In some embodiments, the effective amount comprises 300-500 mg / kg / day of L-serine and 200-400 mg / kg / day of glycine.

[0138] In some embodiments, the genetic disorder is PSATD. In some embodiments, the genetic disorder is NLS2. In some embodiments, reduced activity of PSAT1 or the metabolic pathway in which it functions results in certain diseases, including, but not limited to, ichthyosis, epilepsy, hypertension, retinal degeneration, and macular telangiectasia type 2. Although epilepsy and hypertension are common diseases and only a small proportion of cases associated with such diseases are due to the PSAT1 genotype, detection of a PSAT1 mutation that confers activity below the threshold that causes certain diseases signals the availability of additional treatment options, including dietary supplementation.

[0139] In some embodiments, the subject is a fetus, and the subject is indirectly treated by a carrier of the subject. In some embodiments, the subject is a newborn. In some embodiments, the subject is an infant. In some embodiments, the subject is a pup. In some embodiments, the subject is a juvenile. In some embodiments, the subject is an adult. In some embodiments, one mutation that indicates the presence of a genetic disorder in the subject is selected from the mutations in Table 3.

[0140] In some embodiments, methods are provided for ameliorating, alleviating, or treating a disorder, comprising: receiving diagnostic information comprising the detection of the presence of a DNA variant resulting in at least one mutation in a protein that differs in a subject having the disorder compared to a subject not having the disorder, wherein the presence of the at least one mutation indicates the presence of the disorder in the subject; and administering a therapeutic agent to the subject, wherein the disorder or its symptoms are reduced following administration. In some embodiments, the disorder is associated with reduced activity of serine biosynthesis. In some embodiments, the reduced activity of serine biosynthesis is due to reduced activity of PSAT1. In some embodiments, the DNA variant comprises at least one mutant allele of the PSAT1 gene. In some embodiments, the at least one mutant allele of the PSAT1 gene is hereditary.

[0141] In some embodiments, the disorder comprises ichthyosis. In some embodiments, the disorder comprises epilepsy. In some embodiments, the disorder comprises hypertension. In some embodiments, the disorder comprises retinal degeneration. In some embodiments, the disorder comprises type 2 macular telangiectasia. In some embodiments, the mutant protein comprises an amino acid substitution of at least one amino acid in PSAT1. In some embodiments, PSAT1 is included in a diagnostic panel for ichthyosis, epilepsy, retinal degeneration, hypertension, type 2 macular telangiectasia, or any combination thereof. In some embodiments, a decrease in PSAT1 activity below a threshold indicated by the diagnostic panel is indicative of a disorder. In some embodiments, the therapeutic agent comprises an effective amount of a dietary supplement to improve, alleviate, or treat any of the above disorders. In some embodiments, the dietary supplement comprises L-serine. In some embodiments, an effective amount of L-serine comprises 100 mg / kg / day, 300 mg / kg / day, 400 mg / kg / day, 500 mg / kg / day, or 950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 100-950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 300-950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 400-950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 500-950 mg / kg / day. In some embodiments, the effective amount of L-serine comprises 200 mg / kg / day. In some embodiments, the effective amount of glycine comprises 400 mg / kg / day. In some embodiments, the dietary supplement comprises L-serine and glycine. In some embodiments, the effective amount comprises 500 mg / kg / day of L-serine and 200 mg / kg / day of glycine. In some embodiments, the effective amount comprises 300 mg / kg / day of L-serine and 400 mg / kg / day of glycine. In some embodiments, the effective amount comprises 300-500 mg / kg / day of L-serine and 200-400 mg / kg / day of glycine. [Example]

[0142] Expression constructs encoding protein variants of PSAT1 The human PSAT1 sequence can functionally replace the corresponding SER1 gene in yeast, and this single copy can be stably integrated into the genome under the control of native yeast regulatory sequences for transcription, translation, and mRNA stability of nucleic acids containing the sequence.

[0143] The PSAT1 sequence used was based on the amino acid sequence of PSAT1 isoform 1 (GenBank: NP_478059.1) optimized for yeast expression (hereafter referred to as yPSAT1, GenBank: MN654100, MN654101) (SEQ ID NO: 1538), as shown in Figure 6. At amino acid positions conserved between the yeast and human sequences, the codon sequence of the yeast ortholog SER1 in the S288c reference sequence (R64.2.1 annotation in the Saccharomyces Genome Database) was selected. At non-conserved positions, the codons for the appropriate PSAT1 amino acids were selected to be as similar as possible to the codon usage of the corresponding SER1 codon. For alleles containing frameshift mutations, this same sequence was used up to the frameshift, and the remainder of the open reading frame (ORF) of the human PSAT1 isoform 1 cDNA (GenBank: KJ898759.1) was used.

[0144] For the amino acid substitution mutants, the sequences of SEQ ID NOs: 4 to 1537 shown in Figures 4 and 5 were used.

[0145] For expression in yeast, the yPSAT1 gene was placed under the control of the regulatory elements of the orthologous yeast gene (SER1 promoter and SER1 terminator). The PSAT1_codon_optimized integration plasmid (GenBank accession number MN654100) is described in further detail. 1~6097 = cloning vector pAS19_yPSAT1 1~1336=pUC19 sequence 1337-1577 = S. cerevisiae-derived sequence on the 3' side of the SER1 (YOR184W) terminator 1578–1839 = terminator sequence derived from A. gossypii TEF 1840-2649 = KanR aminoglycoside 3'-phosphotransferase 2650–3083 = promoter sequence from A. gossypii TEF 3084-3546 = S. cerevisiae-derived terminator sequence 3' of the SER1 (YOR184W) stop codon 3547-4659 = yPSAT1 derived from Homo sapiens phosphoserine aminotransferase, codon-optimized for expression in S. cerevisiae 4660-4755 = 5' side of the SER1 (YOR184W) start codon derived from S. cerevisiae 4756~6097=pUC19 sequence [Example]

[0146] Assays to identify highly functional and stable PSAT1 protein variants Yeast growth in the absence of exogenously supplied serine requires phosphoserine aminotransferase activity. Colony growth on minimal media lacking serine is a quantitative measure of enzyme function suitable for high-throughput analysis. A library of all SNP-accessible amino acid substitution mutations spanning the entire length of the human PSAT1 protein was introduced. After single-copy integration into the yeast genome, growth of yeast strains harboring each PSAT1 variant was quantified by assaying growth in media lacking serine using the protocol described herein. Yeast cells expressing wild-type PSAT1 or variant proteins were grown to saturation in rich liquid medium. They were then plated on solid medium (YPG: 1% yeast extract, 2% peptone, 2% glycerol, 2% Bacto Agar) with glycerol as the carbon source, and respiratory-deficient yeast cells were removed. Yeast cells were then plated back into rich liquid medium (YPD: 1% yeast extract, 2% peptone, 2% glucose) and grown to saturation. Yeast cells were then plated in duplicate (n = 3–6) onto solid minimal medium lacking serine (SD: 1.7 g / L Bacto Yeast Nitrogen Base without amino acids or ammonium sulfate, 0.5% ammonium sulfate, 2% glucose, 2% Bacto Agar) and grown at 30°C for 3 days. Plating was performed using a Biomek i7 robot with a V&P 96-pinhead. The DNA sequence of the expression construct present in each strain was determined, and strains with expression constructs containing secondary mutations (in addition to the intended amino acid substitution mutations) were excluded from further consideration. Growth values ​​represent the median of at least one (in most cases more than one) independently derived strain, and each strain was assayed at least three times. The standard error reflects the error associated with these technical and biological replicates.

[0147] Growth scores were used as a measure of the functional impact of variants. Growth was quantified for 95% (1823 of 1914 variants) of the total possible SNP-accessible variants listed in Table 1. Growth values ​​are plotted as points on a scatter plot ordered by amino acid position, as shown in Figure 7. Functional scores below 0.05 in the assay represent alleles with very low activity, such that the strain exhibited little growth. Approximately 3% of variants exhibited growth values ​​in this range of the assay. Approximately 74% of variants fell between 0.05 and the value of the wild-type protein (1.0). The remaining 23% of variants exhibited growth values ​​greater than those conferred by the wild-type protein.

[0148] The results of the assays are expressed as normalized median proliferation values ​​and are shown in Table 1. Protein variants of wild-type SEQ ID NO: 1 that exhibited a proliferation score greater than 1.05 are listed in Table 2. Protein variants of wild-type SEQ ID NO: 1 that exhibited a proliferation score less than 0.99 (e.g., 0.98, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, or lower) are listed in Table 3. Table 1 includes the protein variants listed in Tables 2 and 3.

[0149] [Table 1A]

[0150] [Table 1B]

[0151] [Table 1C]

[0152] [Table 1D]

[0153]

Table 1E

[0154] Table 1F

[0155] Table 1G

[0156]

Table 1H

[0157]

Table 1I

[0158]

Table 1J

[0159] Table 1K

[0160] Table 1L

[0161] Table 1M

[0162]

Table 1N

[0163]

Table 1O

[0164]

Table 1P

[0165]

Table 1Q

[0166]

Table 1R

[0167]

Table 1S

[0168]

Table 1T

[0169]

Table 1U

[0170]

Table 1V

[0171]

Table 1W

[0172] Table 2A

[0173] Table 2B

[0174] Table 3A

[0175] Table 3B

[0176] Table 3C

[0177] Table 3D

[0178] Table 3E

[0179] Table 3F

[0180] Table 3G

[0181]

Table 3H

[0182]

Table 3I

[0183]

Table 3J

[0184] [Table 3K]

[0185] [Table 3L]

[0186] [Table 3M]

[0187] [Table 3N]

[0188] [Table 3O]

[0189] [Table 3P]

[0190] [Table 3Q] [Example]

[0191] Methods for diagnosing and treating PSAT1 deficiency in a subject A subject is identified as likely to have a genetic disorder related to PSATD. The functional status of PSAT1 in the subject is determined. If the subject exhibits at least one mutation resulting in a protein variant with reduced activity and / or stability of PSAT1 listed in Table 3, the subject is diagnosed with PSATD. A composition containing an effective amount of one or more mutants resulting in one or more protein variants with increased activity and / or stability listed in Table 2 is administered to the subject via gene therapy. The subject is continuously monitored for a reduction in symptoms associated with PSATD, and treatment is continued until PSAT1 function is increased and the occurrence and / or symptoms of the disorder are reduced, alleviated, or limited.

[0192] Alternatively, or in conjunction with the above-mentioned therapies, dietary supplements may be administered to the subject. Some non-limiting examples of treatment outcomes are shown in Figure 8. This is reprinted from Shen et al., "Juvenile-onset PSAT1-related neuropathy: A milder phenotype of serine deficiency disorder," Front Genet., August 16, 2022; Vol. 13:949038 (PMID: 36061210). As shown in Figure 8, treatment outcomes 1-3 involved PSAT1 mutations resulting in a condition known as ichthyosis, characterized by scaly, rough, red, dry, and itchy skin. As reported in Figure 8, dietary supplements containing a combination of serine and glycine, or serine alone, were associated with a normal skin phenotype in treatment outcomes 1-3.

[0193] In a non-limiting example involving the PSAT1 gene (PMID: 17436247) variant D100A / G36A_fs*5, two siblings, one older and one younger, were administered dietary supplements. The older sibling was healthy at birth but began experiencing seizures at 7 weeks of age. At 11 weeks of age, the older sibling was administered 500 mg / kg / day of serine and 200 mg / kg / day of glycine for 5 months. However, the older sibling ultimately died at 7 months of age. The younger sibling began receiving serine / glycine treatment at an earlier age than the older sibling. Within 24 hours of birth, the younger sibling was administered 500 mg / kg / day of serine and 200 mg / kg / day of glycine, and follow-up continued until age 3 years. This treatment after variant detection in the younger sibling was associated with a developmentally normal outcome at age 3 years.

[0194] In another non-limiting example involving the detected S43R / S43R variant in the PSAT1 gene (PMID: 26610677), a subject was administered 500 mg / kg / day of serine and 200 mg / kg / day of glycine at age 3 years and continued until age 5.5 years. Serine supplementation at 3 months of age improved the subject's spasticity. However, although the subject was able to establish eye contact and demonstrated a variety of vocalizations, the subject's head circumference was small, there was no significant improvement in psychomotor skills, and the subject remained bedridden.

[0195] In another non-limiting example involving the detected variant A15V / D145M_fs*49 in the PSAT1 gene (PMID: 29269105), a female subject was administered oral serine (500 mg / kg / day) and glycine (200 mg / kg / day) starting at 5 months of age. After starting the serine and glycine supplement, the subject showed improvement in weight gain and irritability. However, the subject continued to have significant developmental delays. While some symptoms of serine deficiency, particularly those resulting from serine deficiency during critical developmental stages, may be irreversible with dietary supplementation, other symptoms (e.g., impaired neurological function or skin repair) may still be correctable with dietary supplementation.

[0196] In another non-limiting example involving the detected variant A15P / A15P in the PSAT1 gene (PMID: 36061210), two young subjects were treated with serine (300 mg / kg / day) and glycine (400 mg / kg / day) for three to five months. The subjects were unrelated and diagnosed at a young age. Both subjects were born healthy; however, one had congenital ichthyosis, and the other developed it at age four (infantile-onset ichthyosis). Both subjects exhibited neuropathic symptoms at ages 19 and 17. This supplementation resulted in resolution of ichthyosis and improved muscle strength in both subjects.

[0197] In another non-limiting example involving the variant A15P / T156M of the PSAT1 gene (PMID: 34089226) in an adult subject (age 38), 100 mg / kg / day of serine was administered, and this supplementation treatment continued until the subject's follow-up at age 40. Serine supplementation in the adult subject, beginning at age 38, resolved ichthyosis and reduced hypertension medication dosage. While supplementation may have halted the progression of neuropathy, it did not appear to significantly reverse it.

[0198] As used herein, section headings are for organizational purposes only and should not be construed as limiting the subject matter described in any way. All literature and similar materials cited in this application, including but not limited to patents, patent applications, papers, books, articles, and Internet web pages, contain the disclosures specifically referenced herein and are expressly incorporated by reference in their entirety for all purposes. If the definition of a term in an incorporated reference is deemed to differ from the definition provided in the present teachings, the definition provided in the present teachings shall control. The word "about" is implicit before temperatures, concentrations, times, etc. discussed in the present teachings, and it will be understood that small, insubstantial deviations are within the scope of the present teachings herein.

[0199] Those skilled in the art will understand that the terms used herein generally, and in the appended claims in particular (e.g., the body of the appended claims), are generally intended to be “open” terms (e.g., the term “including” should be interpreted as “including, but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes, but not limited to,” etc.). Furthermore, those skilled in the art will understand that if a specific number is intended in an introduced claim recitation, such intention will be expressly recited in the claim; in the absence of such recitation, no such intention exists. For example, to aid in understanding, the following appended claims may include the use of the introductory phrases “at least one” and “one or more” to introduce claim recitations. However, the use of such phrases should not be construed as suggesting that introducing a claim recitation with the indefinite article “a” or “an” limits any particular claim that includes such an introduced claim recitation to embodiments that include only one such recitation. The same is true even when the same claim includes the introductory phrase "one or more" or "at least one" and an indefinite article such as "a" or "an" (e.g., "a" and / or "an" should be interpreted to mean "at least one" or "one or more"). The same is true for the use of definite articles used to introduce claim recitations. Additionally, even when a specific number is explicitly recited in an introduced claim recitation, one of skill in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., a minimum recitation of "two recitations," without any other modifier, means at least two recitations, or more than two recitations).Furthermore, when notation similar to "at least one of A, B, and C, etc." is used, such interpretation is generally intended to be the meaning that a person of ordinary skill in the art would understand the notation (e.g., "a system having at least one of A, B, and C" would include, but is not limited to, A only, B only, C only, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). When notation similar to "at least one of A, B, and C, etc." is used, such interpretation is generally intended to be the meaning that a person of ordinary skill in the art would understand the notation (e.g., "a system having at least one of A, B, or C" would include, but is not limited to, A only, B only, C only, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). Furthermore, those skilled in the art should understand that virtually any disjunctive word and / or phrase presenting two or more alternative terms, whether in the specification, claims, or drawings, contemplates the possibility of including one of the terms, either of the terms, or both of the terms. For example, the phrase "A or B" will be understood to include the possibilities of "A" or "B" or "A and B."

[0200] Additionally, when features or aspects of the disclosure are described in terms of a Markush group, one of skill in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0201] Any of the features of the embodiments of the first through eleventh aspects are applicable to all aspects and embodiments identified herein. Furthermore, any of the features of the embodiments of the first through eleventh aspects may be independently combined in any manner, partially or wholly, with other embodiments described herein, for example, one, two, three, or more embodiments may be combined in whole or in part. Furthermore, any of the features of the embodiments of the first through eleventh aspects may be optional with respect to other aspects or embodiments.

Claims

1. The following SEQ ID NOs: a) SEQ ID NO:1408, SEQ ID NO:1409, SEQ ID NO:1410, SEQ ID NO:1411, SEQ ID NO:1412, SEQ ID NO:1413, SEQ ID NO:1414, SEQ ID NO:1415, SEQ ID NO:1416, SEQ ID NO:1417, SEQ ID NO:1418, SEQ ID NO:1419, SEQ ID NO:1420, SEQ ID NO:1421, SEQ ID NO:1422, SEQ ID NO:1423, SEQ ID NO:1424, SEQ ID NO:1425, SEQ ID NO:1426, SEQ ID NO:1427, SEQ ID NO:1428, SEQ ID NO:1429, SEQ ID NO:1430, SEQ ID NO:1431, SEQ ID NO:1432, SEQ ID NO:1433, SEQ ID NO:1434, 34, SEQ ID NO:1435, SEQ ID NO:1436, SEQ ID NO:1437, SEQ ID NO:1438, SEQ ID NO:1439, SEQ ID NO:1440, SEQ ID NO:1441, SEQ ID NO:1442, SEQ ID NO:1443, SEQ ID NO:1444, SEQ ID NO:1445, SEQ ID NO:1446, SEQ ID NO:1447, SEQ ID NO:1448, SEQ ID NO:1449, SEQ ID NO:1450, SEQ ID NO:1451, SEQ ID NO:1452, SEQ ID NO:1453, SEQ ID NO:1454, SEQ ID NO:1455, SEQ ID NO:1456, SEQ ID NO:1457, SEQ ID NO:1458, SEQ ID NO:1459, SEQ ID NO:1460, SEQ ID NO:1461, SEQ ID NO: 1462, SEQ ID NO:1463, SEQ ID NO:1464, SEQ ID NO:1465, SEQ ID NO:1466, SEQ ID NO:1467, SEQ ID NO:1468, SEQ ID NO:1469, SEQ ID NO:1470, SEQ ID NO:1471, SEQ ID NO:1472, SEQ ID NO:1473, SEQ ID NO:1474, SEQ ID NO:1475, SEQ ID NO:1476, SEQ ID NO:1477, SEQ ID NO:1478, SEQ ID NO:1479, SEQ ID NO:1480, SEQ ID NO:1481, SEQ ID NO:1482, SEQ ID NO:1483, SEQ ID NO:1484, SEQ ID NO:1485, SEQ ID NO:1486, SEQ ID NO:1487, SEQ ID NO:1488, SEQ ID NO:1489, SEQ ID NO:1490, SEQ ID NO:1491, SEQ ID NO:1492, SEQ ID NO:1493, SEQ ID NO:1494, SEQ ID NO:1495, SEQ ID NO:1496, SEQ ID NO:1497, SEQ ID NO:1498, SEQ ID NO:149 ... No. 1490, SEQ ID NO. 1491, SEQ ID NO. 1492, SEQ ID NO. 1493, SEQ ID NO. 1494, SEQ ID NO. 1495, SEQ ID NO. 1496, SEQ ID NO. 1497, SEQ ID NO. 1498, SEQ ID NO. 1499, SEQ ID NO. 1500, SEQ ID NO. 1501, SEQ ID NO. 1502, SEQ ID NO. 1503, SEQ ID NO. 1504, SEQ ID NO. 1505, SEQ ID NO. 1506, SEQ ID NO. 1507, SEQ ID NO. 1508, SEQ ID NO. 1509, SEQ ID NO. 1510, SEQ ID NO. 1511, SEQ ID NO. 1512, SEQ ID NO. 1513, SEQ ID NO. 1514, SEQ ID NO. 1515, SEQ ID NO. 1516, SEQ ID NO. 1517,SEQ ID NO: 1518, SEQ ID NO: 1519, SEQ ID NO: 1520, SEQ ID NO: 1521, SEQ ID NO: 1522, SEQ ID NO: 1523, SEQ ID NO: 1524, SEQ ID NO: 1525, SEQ ID NO: 1526, SEQ ID NO: 1527, SEQ ID NO: 1528, SEQ ID NO: 1529, SEQ ID NO: 1530, SEQ ID NO: 1531, SEQ ID NO: 1532, SEQ ID NO: 1533, SEQ ID NO: 1534, SEQ ID NO: 1535, SEQ ID NO: 1536, SEQ ID NO: 1537, or b) a protein having the amino acid sequence of any one of SEQ ID NOs: 4 to 1407.

2. At least 80% identical to SEQ ID NO: 1 (WT sequence) with the following amino acid substitution mutations: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q , K94T, P243A, L239M, K259T, M42I, L221I, K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A1 46T, A15T, L325V, I37T, Y240N, I73M, L29V, K231N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G , I53F, T350P, P144T, G334A, R97M, R5S, N236K, P4T, T242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C , L17Q, E229A, L63V, P4R, I37N, C80G, L357M, A310S, N55T, P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121 A protein having a sequence including one or more of: T, S46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, V157M, P144R, L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F.

3. The following amino acid substitution mutations: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q, K94T, P243A, L239M, K259T, M42I, L2 21I, K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A146T, A15T, L325V, I37T, Y240N, I73M, L2 9V, K231N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G, I53F, T350P, P144T, G334A, R97M, R5S, N23 6K, P4T, T242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C, L17Q, E229A, L63V, P4R, I37N, C80G, L357M, A 310S, N55T, P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121T, S46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, 3. The protein of claim 1 or 2, wherein the protein having at least one of V157M, P144R, L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F exhibits higher activity compared to the WT protein of SEQ ID NO:

1.

4. The following amino acid substitution mutations: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q, K94T, P243A, L239M, K259T, M42I, L221I, K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A146T, A15T, L325V, I37T, Y240N, I73M, L29V, K23 1N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G, I53F, T350P, P144T, G334A, R97M, R5S, N236K, P4T, T 242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C, L17Q, E229A, L63V, P4R, I37N, C80G, L357M, A310S, N55T, P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121T, S46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, V157M, P144R, 4. The protein of any one of claims 1 to 3, wherein the function of the protein having at least one of L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F is enhanced by more than 5% compared to the function of the WT protein of SEQ ID NO:

1.

5. The following amino acid substitution mutations: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q, K94T, P243A, L239M, K259T, M42I, L221I, K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A146T, A15T, L325V, I37T, Y240N, I73M, L29V, K23 1N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G, I53F, T350P, P144T, G334A, R97M, R5S, N236K, P4T, T 242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C, L17Q, E229A, L63V, P4R, I37N, C80G, L357M, A310S, N55T, P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121T, S46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, V157M, P144R, 5. The protein of any one of claims 1 to 4, wherein the function of the protein having at least one of L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F is enhanced by at least 10% compared to the function of the WT protein of SEQ ID NO:

1.

6. Amino acid substitution variants were identified as follows: 2, 3, 4, 5, 7, 8, 12, 15, 17, 22, 24, 26, 29, 31, 37, 42, 46, 48, 49, 53, 55, 56, 63, 68, 71, 72, 73, 74, 80, 85, 93, 94, 95, 97, 106, 109, 110, 116, 117, 121, 125, 132, 133, 136, 138, 144, 146, 148, 157, 169, 174, 180, 193, 198, 203, 219, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 6. The protein of any one of claims 2 to 5, comprising at least one amino acid change at an amino acid position of SEQ ID NO: 1 selected from the group consisting of: 6, 229, 231, 236, 239, 240, 242, 243, 245, 249, 250, 253, 255, 259, 284, 300, 302, 310, 311, 318, 325, 334, 336, 341, 344, 349, 350, 351, 353, 354, 356, 357, 359, 369.

7. Amino acid substitution variants were identified as follows: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 145, 146, 147, 148, 149, 150, 151, 152, 153, 1 54, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 1 85, 186, 187, 188, 189, 190, 191, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217 7, 218, 219, 220, 221, 222, 223, 224, 225, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249 ,250,251,252,253,254,255,256,257,258,259,260,261,262,263,264,265,266,267,268,269,270,271,272,273,274,275,276,277,278,279,280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 6. The protein of any one of claims 2 to 5, comprising at least one amino acid change at an amino acid position of SEQ ID NO: 1 selected from the group consisting of: 34, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370.

8. 7. An expression vector comprising a polynucleotide sequence encoding one or more of the proteins according to any one or more of claims 1 to 6.

9. 9. The expression vector of claim 8, wherein the sequence is a codon-optimized sequence.

10. The following amino acid substitution mutations: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q, K94T, P243A, L239M, K259T, M42I, L221 I, K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A146T, A15T, L325V, I37T, Y240N, I73M, L29V, K231N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G, I53F, T350P, P144T, G334A, R97M, R5S, N236K, P 4T, T242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C, L17Q, E229A, L63V, P4R, I37N, C80G, L357M, A310S, N55T, P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121T, S46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, V157M, 10. The expression vector of claim 9, wherein the polynucleotide sequence comprising one or more of P144R, L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F is enhanced by more than 5% compared to the function of the wild-type protein of SEQ ID NO:

1.

11. 11. The composition of any one of claims 1 to 10, wherein the protein is phosphoserine aminotransferase (PSAT1).

12. 12. A composition according to any one of claims 1 to 11 for use in the treatment of a genetic disorder.

13. 13. The composition of any one of claims 1 to 12, administered to a subject having a genetic disorder in an amount effective to correct the genetic disorder or a symptom thereof in the subject.

14. 14. The composition of claim 12 or 13, wherein the genetic disorder is phosphoserine aminotransferase (PSAT1) deficiency (PSATD).

15. 14. The composition of claim 12 or 13, wherein the genetic disorder is Neuraxova syndrome 2 (NLS2).

16. A method for enhancing the functionality of a PSAT1 protein in a subject, comprising: Administering to said subject a protein or composition according to any one of claims 1 to 14. wherein after administration, PSATD or a symptom thereof is reduced.

17. A method for enhancing the functionality of a PSAT1 protein in a subject, comprising: Administering to said subject a protein or composition according to any one of claims 1 to 13 and 15, wherein NLS2 or a symptom thereof is reduced after administration. A method comprising:

18. 18. The method of claim 16 or claim 17, wherein the subject is a fetus, a newborn, a juvenile, or an adult.

19. 17. The method of claim 16, wherein one or more expression vectors are administered to a subject in an amount effective to correct the PSATD or a symptom thereof in the subject.

20. 18. The method of claim 17, wherein one or more expression vectors are administered to the subject in an amount effective to correct the NLS2 or a symptom thereof in the subject.

21. 18. The method of claim 16 or claim 17, wherein the PSAT1 has been modified to exhibit enhanced function.

22. 18. The method of claim 16 or claim 17, wherein the PSAT1 is replaced or complemented by one or more polynucleotide sequences encoding SEQ ID NOs: 4 to 1537 so as to exhibit enhanced function.

23. The following amino acid substitution mutations: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q, K94T, P243A, L239M, K259T, M42I, L221I , K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A146T, A15T, L325V, I37T, Y240N, I73M, L29V, K2 31N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G, I53F, T350P, P144T, G334A, R97M, R5S, N236K, P4T, T242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C, L17Q, E229A, L63V, P4R, I37N, C80G, L357M, A310S, N55T , P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121T, S46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, V157M, P144R , L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F is enhanced by more than 5% compared to the function of the WT protein of SEQ ID NO:

1.

24. The following amino acid substitution mutations: K110T, P12A, N284T, I73V, V250G, N236S, A3G, K133R, V354F, T132S, I341V, G203A, V72L, F219I, V354L, V250A, I37M, A359V, K94R, L93F, I73F, V193L, V349G, S138G, F74C, S344C, K94Q, K318Q, K94T, P243A, L239M, K259T, M42I, L221I , K356Q, V8F, V7E, I351M, L255M, V72M, D136E, R336S, L22S, N236D, K169Q, A146T, A15T, L325V, I37T, Y240N, I73M, L29V, K2 31N, L63M, K300R, Q369K, D2Y, Y249F, K110E, H125R, E24A, S226A, C245G, I53F, T350P, P144T, G334A, R97M, R5S, N236K, P4T, T242S, Q26R, R97T, N302I, Y148F, T56I, E24K, K169E, D68E, Y249C, L17Q, E229A, L63V, P4R, I37N, C80G, L357M, A310S, N55T , P144Q, A109T, A15S, A85G, D353E, D48G, N180D, K116Q, I121T, S46A, L253S, D31Y, V157L, K116T, F49Y, K311Q, V157M, P144R , L221R, D31A, A95P, K110M, I73L, K356T, A198S, C80W, N55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, I37F, is enhanced by at least 50% compared to the function of the WT protein of SEQ ID NO:

1.

25. 1. A kit for determining a particular mode of treatment for a genetic disorder, comprising: a reference table showing specific amino acid substitutions commonly involved in cellular mechanisms; and One or more vectors comprising the sequences of SEQ ID NOs: 4 to 1537, and optionally specifying at least one or all of SEQ ID NOs: 4 to 1407 Kit including:

26. The reference table includes the following mutations: a) C245R, T242M, G206R, N89Y, G119E, G13R, L40R, I25K, G78V, L63S, G78D, Y148N, G79R, S147Y, I25R, P12L, G203R, I121K, G78A, V39D, H44Y, C152F, T156M, S179L, E155Q, S84R, G11R, D176H, G79W, A99D, G78R, C152R, G119R, D176A, G105V, K200N, Y151D, D176Y, S108P, G197D, Q82R, R45S, G7 8C, A111P, V86D, N154Y, G13S, Q199P, Y148H, R342Q, G159R, K200Q, Y148D, K 200T, E155V, A115D, L182R, A111D, Y101D, G79E, K200R, G197R, C152Y, D176N , E155K, A198D, K200M, R97G, V188D, P14T, P12R, N89K, S38R, Y101N, P87R, G 13C, L255P, T104P, N154D, A109P, L90P, Q82P, N302Y, P14L, P14H, Q199K, T15 6P, T156R, C224W, R61P, S178P, C152W, A112P, T242P, L357R, L90R, H335D, N 302K, W107R, L345R, Y289D, S108L, Y148S, Y101S, K200E, L255R, V174D, G78 S, M251R, N201Y, S147F, W107C, G203D, T242R, V102E, S299R, Q199L, G203V, T242K, S344Y, R213P, D176E, S299I, R342W, Y289S, V174F, W107L, S147P, L33 2W, S344F, M361R, Y70D, R342P, R306P, I278N, Y150N, P243R, C291F, R342L, V212D, G105A, T156A, H158P, W107S, Y148C, M361K, R45M, V157E, C175W, Y289 N, R342G, V193E, Y346N, G340R, A99P, H44L, E155G, V202D, S38I, I194N, Y70 N, S178F, V102G, S178Y, R45G, V72G, N89I, Y101C, Y289H, R45K, N241I, R45T,Y70S、Y346S、I91S、V103G、F10S、G334W、H335L、R213G、I278S、C175R、V149E、Y151N、N89D、I282N、N302H、C291R、Q199E、S179P、G340D、I307N、A115T、I91T、P243H、V72E、S183P、V149G、L332S、S285Y、Q76R、D353V、F305S、A198P、A99T、A324P、H44N、P244R、S272P、M329R、E155A、H44P、G11V、Y346C、L64P、V174G、G119V、C175F、G105E、N154I、T156S、S275P、L270P、A111V、G340C、F181S、S247R、H335P、E155D、H44Q、F191S、I303N、T104K、G119W、Q276P、P14A、L59P、N122I、L17R、A115V、P14S、G11W、S299C、L22W、F364L、R45W、N241Y、Y70C、L75R、N201H、G334V、I121R、P87H、Y230D、V124D、L216R、D187Y、P87L、S179A、G170V、Y289C、A358P、V293M、S178T、G287V、P67L、A205D、W140L、N154T、P292R、V293E、G339V、S275Y、M361T、Y150C、V60E、A343D、W140R、S204Y、F364S、V193G、F83Y、V86G、H335R、A99V、P292Q、Y151S、L321P、R213L、I211S、N89T、E161K、F320C、N180K、S147T、F10I、G263V、P292A、C245Y、I282S、P166L、F364C、F288C、G340S、P166T、I282T、P292S、I303F、E161V、I278T、A111T、G197V、A153P、P166H、C245F、R213C、L64R、M361V、H335Q、T104R、H335N、S344P、S285C、P166S、L216P、F320I、W140G、V338E、G206V、V66F、L228P、F364V、F195S、S43R、Q233P、W140C、Y279H、V188G、R61W、R298I、A146D、D283H、L63W、I303S、M42R、C224Y、S285T、V209E、F320L、F364I、V174L、L357Q、R298S、I307S、V103E、C175Y、E317D、R298T、A106T、I341N、L216Q、N180Y、E317G、T139P、V172G、D163V、A112E、S285A、A324D、E366V、I282F、H158Y、F181C、Y279D、A359P、F191V、N347Y、I281F、S275T、K274I、P12T、Y279F、I303M、G92V、P292L、P166A、R213S、V293A、M177K、V186G、R298K、D163Y、N328I、I134L、V210E、V60G、R298G、V202G、Y279S、D353A、G206E、V21E、I134T、N89S、A264D、C80R、T277P、D100Y、F49S、Q233R、F320Y、F162V、G197S、G262V、Y230S、I134N、A15D、K94I、H368R、Y70H、I134S、I307T、D353Y、A358D、E114K、V290G、V160M、V160G、F118S、Q286P、S204P、V124G、T56R、H158R、M329I、N347D、E161D、F288S、F162I、I248S、K333N、N180H、L182Q、R61L、F10V、T104I、A15P、F288V、K33I、W257S、K274E、M267K、N302D、I281N、F191L、G79A、N347T、D100G、I211T、F305L、F83C、E366K、I303T、L182P、C291S、V186E、V293L、M329V、L316S、I281S、F162S、D163H、D100V、F288I、M267R、A348G、D283V、A99G、S204F、P67R、R5G、P225L、Y230H、L17M、V124F、F162C、V102L、I134F、I278F、G77E、V21G、V160A、V209G、F288L、F360S、C224R、F181L、Y230C、N89H、S189F、I282M、F191C、D314A、L75V、F10C、S178C、S20L、T56P、P67A、R306G、N69I、I258T、I134M、K190E、E57K、S179T、P126A、F305Y、H158L、H335Y、I341L、F305V、E317K、T277R、F83I、P87S、P244L、F83V、F49C、D187G、D283G、L316V、P4L、D100A、K190N、A15V、K356R、A220D、E280K、S108T、Y32D、G263R、V354G、S84T、M361L、I307F、V250F、P225H、F74S、E268V、E114A、E366A、S299T、N284I、A234P、Y101H、D353G、V168D、G218W、P67T、A106P、C152G、P18Q、I194S、C98F、D68Y、A99S、L90F、V202A、N54I、V103M、V290L、P67Q、Y151C、F49L、N58K、Y230N、L327P、V338G、G203C、V102A、T242A、R61Q、F49V、V354A、P135L、K190T、I194F、A109V、H125D、D163A、S46P、Y101F、S337P、K51Q、L59V、N297T、V124A、I307M、A266P、E280V、G159A、C224F、V149A、K116R、N154H、G263D、I52M、Y240D、Y346H、E352V、E229D、F181V、V86F、G197A、D187V、A106G、N54T、F162L、F181I、K333Q、D136A、P67S、D136G、G339E、D68A、W257C、G262A、K274R、S84I、I248N、G159C、K71N、M42V、V349D、I282L、N328Y、L228R、Q76E、V212G、S84C、R222G、E326Q、V35F、S43I、V7G、S183T、Q76H、K190R、V66L、S130R、Q276E、F305I、L40I、G287R、T104S、A343G、D167V、G235V、A220V、V254D、G77V、V157G、R213H、L239V、S238P、D187H、N302S、S275C、G339A、L217M、A196T、A220P、T208I、D100E、S247C、A264G、E62V、E161Q、V290D、S183C、Q6H、D167Y、H19L、I165L、S299G、S20P、Y32F、C98W、C175S、A153T、I258S、E161G、P137R、N284Y、L216M、D187A、A343S、Y279N、K51R、I123F、V21A、S130T、G36C、G308D、V188F、P126R、D215V、K33R、V207F、D100H、S183Y、N328S、I91V、K127R、L217R、I91F、N143K、N201K、V21L、N297D、G262R、G312V、P135T、N69D、N55I、V60A、N69Y、V250D、I165R、K300T、N201I、K323N、I53N、S20T、Q276L、K190Q、L128V、S204C、D187N、F83L、D215N、D353N、H19Q、G119A、E57D、G334E、A265T、Y32H、I73S、S108A、V35I、G81S、V186M、I165K、S331T、L23I、E317Q、V39L、T139S、P126L、H19D、P18A、V212F、G81C、H158Q、L173P、A348V、L316F、N180I、E57V、G235E、P304A、D145V、E24D、A112T、K323R、Q296P、S43G、I351V、N69K、G334R、R222P、T120S、E28A、E366G、V168I、V72A、D353H、I341S、L93S、L142H、T139N、D31E、P137T、F181Y、N309D、V103A、L75P、A85V、H125P、R222L、D163E、D215H、K333E、G129W、N309I、Q369L、G170A、R306C、S247I、V103L、P18R、N347H、E57G、I25M、P244S、W257G、L325H、I273S、I123V、K300E、I53S、Q82L、L327H、L270R、G36V、A266V、V21M、I282V、K300M、S271G、S189P、A50G、C224S、V124L、I52L、T277A、E62Q、K323Q、D313Y、A171S、N328K、I52T、I165M、K27Q、K16R、E24G、F288Y、L255Q、A220T、F195C、T132R、Q286R、G92A、V188A、Q369R、H125L、E256Q、T56A、E268K、P137Q、N54D、L59I、E57A、S147A、K94E、K51M、S237N、V60L、L253M、N236H、G235A、I211F、K16Q、A65V、D283A、D163G、G287A、P225A、K127N、K33E、F364Y、L59H、V39F、A109G、A171V、V227D、T277I、L217Q 、M177T、H19P、F164S、G263C、E223V、L216V、D48N、L217V、E229Q、P295L、E11 3V、F10L、F164I、I303V、S38G、K323E、N58T、N297H、M267T、R5T、V186A、N58I、 K27T、A146P、G308A、K133I、P244T、G252V、C224G、G92S、G81V、V202L、V168L 、V290I、T104A、P185A、Y240H、E28Q、L345M、G36R、A198V、S272T、K333I、I341 M、G263S、E280G、K274T、I165T、D2A、Q296H、A109D、E114V、Q355H、K318E、V2 54I、D322V、A153S、C98Y、D215Y、I73N、N241T、N141H、K363N、V35L、T120P、I2 81V、P225S、V212I、A109S、D68G、K274Q、L221P、D215G、L30F、E24V、L64V、D100N、M177V、L128F、G92C、V160L、A198G、E28D、P87T、S147C、E268A、R5M、V290 A、A85T、P225T、S247G、R319G、H19R、L365W、I307V、L142V、E223K、L128R、S183A、T132I、W257R、G312R、P135A、D313A、Q276K、P87A、S204A、S38C、K362R、V 207L、V250I、S331Y、T139A、A3D、A65D、E294G、H368P、A146G、S237I、T139I、F164Y、N54Y、K184N、R61G、V102M、V60M、I307L、I52N、K363R、D187E、E317A、V 149L、E62A、S130C、A265S、D145E、V202I、N122H、Q82E、L64Q、M267L、L59F、S 130G、I248F、D145A、K16E、S237G、D167E、Y150H、Q286L、N297Y、A95E、K300N、S247T、K33Q、I123T、K269E、M251T、A153G、Y230F、P135R、F164V、A171P、D145Y、N328T、L270V、I25V、I121L、S189C、A266S、V35G、C98S、M301T、V212L、V7M、D167N、I121M、T350I、P295T、A358V、K190M、P126S、Q276H、G287E、A3S、D215E、K127T、N58Y、F164C、Q296E、F118Y、L142I、L40V、G218A、G96V、M267V、N284D、L88I、D167H、V8D、P185L、P244A、T277S、A310V、R5K、L330M、F164L、L217P、A343P、N141S、N58H、D31V、G312E、L270F、S237C、E268G、A324S、F74V、D314Y、D2V、L370I、N55S、I25L、E366D、L345P、A265P、L17V、A315P、K333R、K363E、V254L、Q369P、V86A、T120I、V254A、D48V、Y70F、E268Q、N261T、N309T、P244Q、K269M、G92D、C152S、N141Y、S238Y、G218R、N9S、V232L、I258L、N284S、E256D、S84N、Q369H、A146S、K231E、V254G、H368D、L253W、E256V、G262E、R306S、Q233K、I278M、E294V、V174A、N261I、S43C、K71R、S138C、L325P、D322G、D2G、A358T、A106V、K269N、V232E、E114G、F360I、P295A、L253V、V210G、V124I、I123M、V207G、I278V、K318I、F246S、E113D、A3T、K27M、S46L、L327R、G218E、L88S、A65G、Q286E、Q296K、K184R、P18S、Q6R、F360Y、D48A、A146V、I273L、L332V、N141D、V168A、V232G、I53V、L59R、T277K、N328D、Q233H、S272C、I52S、L75Q、S130N、L327F、N141K、D322N、S138I、D314H、Y151H、L142R、K274N、G96A、L40F、A234S、K184E、N58S、L182V、E294K、M301K、P12H、L327V、K362T、I91L、G81A、 I73T、I351S、K362Q、L40H、L330V、Q26L、N309Y、V212A、A266D、G308S、F360C 、A111G、D68H、S247N、A50D、S331P、K363Q、E326A、K362N、K33N、E229V、Y249D、N261K、K27N、G170R、K311N、A220S、P4A、I278L、A112V、I273N、T132P、L221 F、S237T、D214H、L142P、G218V、G36D、I341F、E268D、L142F、G308R、A315S、D 31N、A264P、R306L、S47A、G34V、I211V、I258N、E223Q、N284H、A50S、P295S、Q6 E、A358G、L321H、D214A、A205S、V188I、Y240F、I273M、F118L、D313E、D314G、 G36A、I273V、N122T、R319I、A65S、C291G、G34R、F219Y、S271R、Q276R、M42L、G 339R、E57Q、V168G、P18L、K169N、D322E、A310P、E113A、V66I、K259I、S331A、N260H、L29S、I123L、I52F、S337T、M367R、L90V、K231Q、F219V、K127Q、D214G 、P126H、A324T、I194T、Y249S、Q76P、I37S、D145N、G77A、D314E、E294Q、K127E、D313G、S138N、A324G、D48H、Y240C、D68N、G129A、I281L、V202F、L365S、D32 2A、E280D、I273F、K110R、K27E、E161A、N201S、F219L、V193A、L239W、K363T、K269T、I248V、N143D、D31G、Q355L、E280Q、A196V、L93W、W257L、K323T、Q82H、 V250L、S272Y、G252D、D313H、P304R、L270H、Q6P、A264S、I211M、L128I、N309 K、V86I、N122D、N55Y、M367V、H19N、V172E、H368N、K184T、K16T、D136N、R97K、A266T、R336T、N260I、N260Y、D2N、S272F、I351L、M329L、L321V、A343V、G129E、N54K、K110T、P12A、N284T、I73V、V250G、N236S、A3G、K133R、V354F、T132S、I341V、G203A、V72L、F219I、V354L、V250A、I37M、A359V、K94R、L9 3F、I73F、V193L、V349G、S138G、F74C、S344C、K94Q、K318Q、K94T、P243A、L239M、K259T、M42I、L221I、K356Q、V8F、V7E、I351M、L255M、V72M、D136E、R336S、L22S、N236D、K169Q、A146T、A15T、L325V、I37T、Y240N、I73M、L29 V、K231N、L63M、K300R、Q369K、D2Y、Y249F、K110E、H12 5R、E24A、S226A、C245G、I53F、T350P、P144T、G334A、R9 7M、R5S、N236K、P4T、T242S、Q26R、R97T、N302I、Y148F 、T56I、E24K、K169E、D68E、Y249C、L17Q、E229A、L63V、P 4R、I37N、C80G、L357M、A310S、N55T、P144Q、A109T、A1 5S、A85G、D353E、D48G、N180D、K116Q、I121T、S46A、L25 3S、D31Y、V157L、K116T、F49Y、K311Q、V157M、P144R、L221R、D31A、A95P、K110M、I73L、K356T、A198S、C80W、N、 26. The kit of claim 25, comprising the following mutations: 55H, A106D, I121V, K117N, I37L, K117M, K117R, V174I, K117Q, P4H, N55K, K116E, K71Q, and I37F; or b) a list of only the mutations in a) that are also listed in Table 3.

27. 27. The kit of claim 25 or claim 26, wherein the genetic disorder is PSATD.

28. 27. The kit of claim 25 or claim 26, wherein the genetic disorder is NLS2.

29. 1. A method for identifying a subject having a genetic disorder, comprising: detecting the presence of at least one mutation in a protein that differs in said subject with a genetic disorder compared to a subject without said disorder, wherein the presence of said at least one mutation indicates the presence of a genetic disorder in said subject. A method comprising:

30. 1. A method for ameliorating, alleviating, or treating a genetic disorder, comprising: detecting the presence of at least one mutation in a protein that differs in a subject with a genetic disorder compared to a subject without said disorder, wherein the presence of said at least one mutation indicates the presence of a genetic disorder in said subject; and Administering to said subject a protein or composition according to any one of claims 1 to 13, wherein said genetic disorder or symptoms thereof are reduced after administration. A method comprising:

31. the mutations are C245R, T242M, G206R, N89Y, G119E, G13R, L40R, I25K, G78V, L63S, G78D, Y148N, G79R, S147Y, I25R, P12L, G203R, I121K, G78A, V39D, H44Y, C152F, T156M, S179L, E155Q, S84R, G11R, D176H, G79W, A99D, G78R, C152R, G119R, D 176A, G105V, K200N, Y151D, D176Y, S108P, G197D, Q82R, R45S, G78C, A111P, V 86D, N154Y, G13S, Q199P, Y148H, R342Q, G159R, K200Q, Y148D, K200T, E155V , A115D, L182R, A111D, Y101D, G79E, K200R, G197R, C152Y, D176N, E155K, A19 8D, K200M, R97G, V188D, P14T, P12R, N89K, S38R, Y101N, P87R, G13C, L255P, T104P, N154D, A109P, L90P, Q82P, N302Y, P14L, P14H, Q199K, T156P, T156R, C 224W, R61P, S178P, C152W, A112P, T242P, L357R, L90R, H335D, N302K, W107R , L345R, Y289D, S108L, Y148S, Y101S, K200E, L255R, V174D, G78S, M251R, N20 1Y, S147F, W107C, G203D, T242R, V102E, S299R, Q199L, G203V, T242K, S344Y , R213P, D176E, S299I, R342W, Y289S, V174F, W107L, S147P, L332W, S344F, M3 61R, Y70D, R342P, R306P, I278N, Y150N, P243R, C291F, R342L, V212D, G105A , T156A, H158P, W107S, Y148C, M361K, R45M, V157E, C175W, Y289N, R342G, V19 3E, Y346N, G340R, A99P, H44L, E155G, V202D, S38I, I194N, Y70N, S178F, V10 2G, S178Y, R45G, V72G, N89I, Y101C, Y289H, R45K, N241I, R45T, Y70S, Y346S,I91S、V103G、F10S、G334W、H335L、R213G、I278S、C175R、V149E、Y151N、N89D、I282N、N302H、C291R、Q199E、S179P、G340D、I307N、A115T、I91T、P243H、V72E、S183P、V149G、L332S、S285Y、Q76R、D353V、F305S、A198P、A99T、A324P、H44N、P244R、S272P、M329R、E155A、H44P、G11V、Y346C、L64P、V174G、G119V、C175F、G105E、N154I、T156S、S275P、L270P、A111V、G340C、F181S、S247R、H335P、E155D、H44Q、F191S、I303N、T104K、G119W、Q276P、P14A、L59P、N122I、L17R、A115V、P14S、G11W、S299C、L22W、F364L、R45W、N241Y、Y70C、L75R、N201H、G334V、I121R、P87H、Y230D、V124D、L216R、D187Y、P87L、S179A、G170V、Y289C、A358P、V293M、S178T、G287V、P67L、A205D、W140L、N154T、P292R、V293E、G339V、S275Y、M361T、Y150C、V60E、A343D、W140R、S204Y、F364S、V193G、F83Y、V86G、H335R、A99V、P292Q、Y151S、L321P、R213L、I211S、N89T、E161K、F320C、N180K、S147T、F10I、G263V、P292A、C245Y、I282S、P166L、F364C、F288C、G340S、P166T、I282T、P292S、I303F、E161V、I278T、A111T、G197V、A153P、P166H、C245F、R213C、L64R、M361V、H335Q、T104R、H335N、S344P、S285C、P166S、L216P、F320I、W140G、V338E、G206V、V66F、L228P、F364V、F195S、S43R、Q233P、W140C、Y279H、V188G、R61W、R298I、A146D、D283H、L63W、I303S、M42R、C224Y、S285T、V209E、F320L、F364I、V174L、L357Q、R298S、I307S、V103E、C175Y、E317D、R298T、A106T、I341N、L216Q、N180Y、E317G、T139P、V172G、D163V、A112E、S285A、A324D、E366V、I282F、H158Y、F181C、Y279D、A359P、F191V、N347Y、I281F、S275T、K274I、P12T、Y279F、I303M、G92V、P292L、P166A、R213S、V293A、M177K、V186G、R298K、D163Y、N328I、I134L、V210E、V60G、R298G、V202G、Y279S、D353A、G206E、V21E、I134T、N89S、A264D、C80R、T277P、D100Y、F49S、Q233R、F320Y、F162V、G197S、G262V、Y230S、I134N、A15D、K94I、H368R、Y70H、I134S、I307T、D353Y、A358D、E114K、V290G、V160M、V160G、F118S、Q286P、S204P、V124G、T56R、H158R、M329I、N347D、E161D、F288S、F162I、I248S、K333N、N180H、L182Q、R61L、F10V、T104I、A15P、F288V、K33I、W257S、K274E、M267K、N302D、I281N、F191L、G79A、N347T、D100G、I211T、F305L、F83C、E366K、I303T、L182P、C291S、V186E、V293L、M329V、L316S、I281S、F162S、D163H、D100V、F288I、M267R、A348G、D283V、A99G、S204F、P67R、R5G、P225L、Y230H、L17M、V124F、F162C、V102L、I134F、I278F、G77E、V21G、V160A、V209G、F288L、F360S、C224R、F181L、Y230C、N89H、S189F、I282M、F191C、D314A、L75V、F10C、S178C、S20L、T56P、P67A、R306G、N69I、I258T、I134M、K190E、E57K、S179T、P126A、F305Y、H158L、H335Y、I341L、F305V、E317K、T277R、F83I、P87S、P244L、F83V、F49C、D187G、D283G、L316V、P4L、D100A、K190N、A15V、K356R、A220D、E280K、S108T、Y32D、G263R、V354G、S84T、M361L、I307F、V250F、P225H、F74S、E268V、E114A、E366A、S299T、N284I、A234P、Y101H、D353G、V168D、G218W、P67T、A106P、C152G、P18Q、I194S、C98F、D68Y、A99S、L90F、V202A、N54I、V103M、V290L、P67Q、Y151C、F49L、N58K、Y230N、L327P、V338G、G203C、V102A、T242A、R61Q、F49V、V354A、P135L、K190T、I194F、A109V、H125D、D163A、S46P、Y101F、S337P、K51Q、L59V、N297T、V124A、I307M、A266P、E280V、G159A、C224F、V149A、K116R、N154H、G263D、I52M、Y240D、Y346H、E352V、E229D、F181V、V86F、G197A、D187V、A106G、N54T、F162L、F181I、K333Q、D136A、P67S、D136G、G339E、D68A、W257C、G262A、K274R、S84I、I248N、G159C、K71N、M42V、V349D、I282L、N328Y、L228R、Q76E、V212G、S84C、R222G、E326Q、V35F、S43I、V7G、S183T、Q76H、K190R、V66L、S130R、Q276E、F305I、L40I、G287R、T104S、A343G、D167V、G235V、A220V、V254D、G77V、V157G、R213H、L239V、S238P、D187H、N302S、S275C、G339A、L217M、A196T、A220P、T208I、D100E、S247C、A264G、E62V、E161Q、V290D、S183C、Q6H、D167Y、H19L、I165L、S299G、S20P、Y32F、C98W、C175S、A153T、I258S、E161G、P137R、N284Y、L216M、D187A、A343S、Y279N、K51R、I123F、V21A、S130T、G36C、G308D、V188F、P126R、D215V、K33R、V207F、D100H、S183Y、N328S、I91V、K127R、L217R、I91F、N143K、N201K、V21L、N297D、G262R、G312V、P135T、N69D、N55I、V60A、N69Y、V250D、I165R、K300T、N201I、K323N、I53N、S20T、Q276L、K190Q、L128V、S204C、D187N、F83L、D215N、D353N、H19Q、G119A、E57D、G334E、A265T、Y32H、I73S、S108A、V35I、G81S、V186M、I165K、S331T、L23I、E317Q、V39L、T139S、P126L、H19D、P18A、V212F、G81C、H158Q、L173P、A348V、L316F、N180I、E57V、G235E、P304A、D145V、E24D、A112T、K323R、Q296P、S43G、I351V、N69K、G334R、R222P、T120S、E28A、E366G、V168I、V72A、D353H、I341S、L93S、L142H、T139N、D31E、P137T、F181Y、N309D、V103A、L75P、A85V、H125P、R222L、D163E、D215H、K333E、G129W、N309I、Q369L、G170A、R306C、S247I、V103L、P18R、N347H、E57G、I25M、P244S、W257G、L325H、I273S、I123V、K300E、I53S、Q82L、L327H、L270R、G36V、A266V、V21M、I282V、K300M、S271G、S189P、A50G、C224S、V124L、I52L、T277A、E62Q、K323Q、D313Y、A171S、N328K、I52T、I165M、K27Q、K16R、E24G、F288Y、L255Q、A220T、F195C、T132R、Q286R、G92A、V188A、Q369R、H125L、E256Q、T56A、E268K、P137Q、N54D、L59I、E57A、S147A、K94E、K51M、S237N、V60L、L253M、N236H、G235A、I211F、K16Q、A65V、D283A、D163G、G287A、P225A、K127N、K33E、F364Y、L59H、V39F、A109G、A171V、V227D、T277I、L217Q、M177T 、H19P、F164S、G263C、E223V、L216V、D48N、L217V、E229Q、P295L、E113V、F10 L、F164I、I303V、S38G、K323E、N58T、N297H、M267T、R5T、V186A、N58I、K27T、A 146P、G308A、K133I、P244T、G252V、C224G、G92S、G81V、V202L、V168L、V290I 、T104A、P185A、Y240H、E28Q、L345M、G36R、A198V、S272T、K333I、I341M、G263 S、E280G、K274T、I165T、D2A、Q296H、A109D、E114V、Q355H、K318E、V254I、D3 22V、A153S、C98Y、D215Y、I73N、N241T、N141H、K363N、V35L、T120P、I281V、P2 25S、V212I、A109S、D68G、K274Q、L221P、D215G、L30F、E24V、L64V、D100N、M177V、L128F、G92C、V160L、A198G、E28D、P87T、S147C、E268A、R5M、V290A、A85T 、P225T、S247G、R319G、H19R、L365W、I307V、L142V、E223K、L128R、S183A、T1 32I、W257R、G312R、P135A、D313A、Q276K、P87A、S204A、S38C、K362R、V207L、V 250I、S331Y、T139A、A3D、A65D、E294G、H368P、A146G、S237I、T139I、F164Y、N54Y、K184N、R61G、V102M、V60M、I307L、I52N、K363R、D187E、E317A、V149L、E 62A、S130C、A265S、D145E、V202I、N122H、Q82E、L64Q、M267L、L59F、S130G、I 248F、D145A、K16E、S237G、D167E、Y150H、Q286L、N297Y、A95E、K300N、S247T、K33Q、I123T、K269E、M251T、A153G、Y230F、P135R、F164V、A171P、D145Y、N328T、L270V、I25V、I121L、S189C、A266S、V35G、C98S、M301T、V212L、V7M、D167N、I121M、T350I、P295T、A358V、K190M、P126S、Q276H、G287E、A3S、D215E、K127T、N58Y、F164C、Q296E、F118Y、L142I、L40V、G218A、G96V、M267V、N284D、L88I、D167H、V8D、P185L、P244A、T277S、A310V、R5K、L330M、F164L、L217P、A343P、N141S、N58H、D31V、G312E、L270F、S237C、E268G、A324S、F74V、D314Y、D2V、L370I、N55S、I25L、E366D、L345P、A265P、L17V、A315P、K333R、K363E、V254L、Q369P、V86A、T120I、V254A、D48V、Y70F、E268Q、N261T、N309T、P244Q、K269M、G92D、C152S、N141Y、S238Y、G218R、N9S、V232L、I258L、N284S、E256D、S84N、Q369H、A146S、K231E、V254G、H368D、L253W、E256V、G262E、R306S、Q233K、I278M、E294V、V174A、N261I、S43C、K71R、S138C、L325P、D322G、D2G、A358T、A106V、K269N、V232E、E114G、F360I、P295A、L253V、V210G、V124I、I123M、V207G、I278V、K318I、F246S、E113D、A3T、K27M、S46L、L327R、G218E、L88S、A65G、Q286E、Q296K、K184R、P18S、Q6R、F360Y、D48A、A146V、I273L、L332V、N141D、V168A、V232G、I53V、L59R、T277K、N328D、Q233H、S272C、I52S、L75Q、S130N、L327F、N141K、D322N、S138I、D314H、Y151H、L142R、K274N、G96A、L40F、A234S、K184E、N58S、L182V、E294K、M301K、P12H、L327V、K362T、I91L、G81A、I73T、I 351S、K362Q、L40H、L330V、Q26L、N309Y、V212A、A266D、G308S、F360C、A111G 、D68H、S247N、A50D、S331P、K363Q、E326A、K362N、K33N、E229V、Y249D、N261K、K27N、G170R、K311N、A220S、P4A、I278L、A112V、I273N、T132P、L221F、S237 T、D214H、L142P、G218V、G36D、I341F、E268D、L142F、G308R、A315S、D31N、A2 64P、R306L、S47A、G34V、I211V、I258N、E223Q、N284H、A50S、P295S、Q6E、A358 G、L321H、D214A、A205S、V188I、Y240F、I273M、F118L、D313E、D314G、G36A、I 273V、N122T、R319I、A65S、C291G、G34R、F219Y、S271R、Q276R、M42L、G339R、E 57Q、V168G、P18L、K169N、D322E、A310P、E113A、V66I、K259I、S331A、N260H、L29S、I123L、I52F、S337T、M367R、L90V、K231Q、F219V、K127Q、D214G、P126H 、A324T、I194T、Y249S、Q76P、I37S、D145N、G77A、D314E、E294Q、K127E、D313G、S138N、A324G、D48H、Y240C、D68N、G129A、I281L、V202F、L365S、D322A、E28 0D、I273F、K110R、K27E、E161A、N201S、F219L、V193A、L239W、K363T、K269T、I248V、N143D、D31G、Q355L、E280Q、A196V、L93W、W257L、K323T、Q82H、V250L、 S272Y、G252D、D313H、P304R、L270H、Q6P、A264S、I211M、L128I、N309K、V86I 、N122D、N55Y、M367V、H19N、V172E、H368N、K184T、K16T、D136N、R97K、A266T、The method of claim 29 or 30, wherein the nucleotide sequence is selected from the group consisting of R336T, N260I, N260Y, D2N, S272F, I351L, M329L, L321V, A343V, G129E, and N54K.

32. 32. The method of any one of claims 29 to 31, wherein the genetic disorder is PSATD.

33. 31. The method of claim 29 or 30, wherein the one mutation that is indicative of the presence of a genetic disorder in the subject is selected from the mutations in Table 3.

34. 34. The method of claim 33, wherein the engineered protein administered to the subject comprises one or more of the mutations in Table 2.

35. 1. A method for ameliorating, alleviating, or treating a genetic disorder, comprising: receiving diagnostic information comprising the detection of the presence of a DNA variant that results in at least one mutation in a protein that differs in a subject with a genetic disorder compared to a subject without the disorder, wherein the presence of the at least one mutation indicates the presence of the genetic disorder in the subject; and administering a therapeutic agent to the subject, wherein the genetic disorder or symptoms thereof are reduced after administration. A method comprising:

36. 36. The method of claim 35, wherein the therapeutic agent comprises an effective amount of a dietary supplement.

37. 37. The method of claim 36, wherein the dietary supplement comprises L-serine.

38. 38. The method of claim 37, wherein the effective amount comprises 100 to 950 mg / kg / day of L-serine.

39. 38. The method of claim 37, wherein the effective amount comprises 400 to 950 mg / kg / day of L-serine.

40. 38. The method of claim 37, wherein the effective amount comprises 500 mg / kg / day of L-serine.

41. 37. The method of claim 36, wherein the dietary supplement comprises glycine.

42. 42. The method of claim 41, wherein the effective amount comprises 200 mg / kg / day of glycine.

43. 42. The method of claim 41, wherein the effective amount comprises 400 mg / kg / day of glycine.

44. 42. The method of claim 41, wherein the effective amount comprises 200 to 400 mg / kg / day of glycine.

45. 37. The method of claim 36, wherein the dietary supplement comprises L-serine and glycine.

46. 46. ​​The method of claim 45, wherein the effective amounts comprise 500 mg / kg / day of L-serine and 200 mg / kg / day of glycine.

47. 46. ​​The method of claim 45, wherein the effective amounts comprise 300 mg / kg / day of L-serine and 400 mg / kg / day of glycine.

48. The effective amount is 300–500 mg / kg / day of L-serine, and 200–400 mg / kg / day of glycine 46. ​​The method of claim 45, comprising:

49. 49. The method of any one of claims 35 to 48, wherein the genetic disorder is PSATD.

50. 49. The method of any one of claims 35 to 48, wherein the genetic disorder is NLS2.

51. 49. The method of any one of claims 35 to 48, wherein the one mutation indicative of the presence of the genetic disorder in the subject is selected from the mutations in Table 3.

52. 1. A method for ameliorating, alleviating, or treating a disorder, comprising: receiving diagnostic information comprising the detection of the presence of a DNA variant that results in at least one mutation in a protein that differs in a subject with a disorder compared to a subject without said disorder, wherein the presence of said at least one mutation indicates the presence of said disorder in said subject; administering a therapeutic agent to the subject, wherein the disorder or symptoms thereof are reduced after administration. Including, The method, wherein the disorder is associated with reduced activity of serine biosynthesis.

53. 53. The method of claim 52, wherein the disorder comprises ichthyosis.

54. 53. The method of claim 52, wherein the disorder comprises epilepsy.

55. 53. The method of claim 52, wherein the disorder comprises hypertension.

56. 53. The method of claim 52, wherein the disorder comprises retinal degeneration.

57. 53. The method of claim 52, wherein the disorder comprises type 2 macular telangiectasia.

58. 58. The method of any one of claims 52 to 57, wherein the mutant protein comprises an amino acid substitution of at least one amino acid of WT PSAT1.

59. 59. The method of any one of claims 52 to 58, wherein the therapeutic agent comprises an effective amount of a dietary supplement.

60. 60. The method of claim 59, wherein the dietary supplement comprises L-serine.

61. 61. The method of claim 60, wherein the effective amount comprises 100 to 950 mg / kg / day of L-serine.

62. 61. The method of claim 60, wherein the effective amount comprises 400 to 950 mg / kg / day of L-serine.

63. 61. The method of claim 60, wherein the effective amount comprises 500 mg / kg / day of L-serine.

64. 60. The method of claim 59, wherein the dietary supplement comprises glycine.

65. 65. The method of claim 64, wherein the effective amount comprises 200 mg / kg / day of glycine.

66. 65. The method of claim 64, wherein the effective amount comprises 400 mg / kg / day of glycine.

67. 65. The method of claim 64, wherein the effective amount comprises 200 to 400 mg / kg / day of glycine.

68. 60. The method of claim 59, wherein the dietary supplement comprises L-serine and glycine.

69. 69. The method of claim 68, wherein the effective amounts comprise 500 mg / kg / day of L-serine and 200 mg / kg / day of glycine.

70. 69. The method of claim 68, wherein the effective amounts comprise 300 mg / kg / day of L-serine and 400 mg / kg / day of glycine.

71. The effective amount is 300–500 mg / kg / day of L-serine, and 200–400 mg / kg / day of glycine 69. The method of claim 68, comprising:

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