Compositions and methods for the treatment of autosomal dominant polycystic kidney disease

Abstract

The present disclosure generally relates to compositions and methods for the treatment of autosomal dominant polycystic kidney disease type 2 (ADPKD2). In particular, the present disclosure provides nucleic acid molecules comprising a polynucleotide encoding a polycystin‑2 (PC2) protein, as well as vectors and recombinant AAV particles comprising such nucleic acid molecules, and their use in the treatment of ADPKD2.

Description

Attorney Docket No. TORQ-012 / 02WO 339010-2071COMPOSITIONS AND METHODS FOR THE TREATMENT OF AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASECROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and the priority from U. S. Provisional Application No. 63 / 771,155, filed on March 13, 2025, the disclosure of which is incorporated by reference in its entirety.FIELD OF THE DISCLOSURE

[0002] The present disclosure generally relates to compositions and methods for the treatment of autosomal dominant polycystic kidney disease (ADPKD), particularly ADPKD type 2 (ADPKD2) caused by mutations in the PKD2 gene.INCORPORATION OF THE SEQUENCE LISTING

[0003] The contents of the electronic sequence listing (TORQ_012_02WO_SeqList_ST26.xml; Size: 592,417 bytes; and Date of Creation: March 11, 2026) are herein incorporated by reference in its entirety.BACKGROUND

[0004] Autosomal dominant polycystic kidney disease (ADPKD) is a genetic disorder characterized by the formation of fluid-filled cysts in the kidneys. These cysts can grow over time and may lead to kidney enlargement and loss of kidney function.

[0005] The PKD2 gene encodes polycystin-2 (PC2), a multifunctional, transmembrane protein that plays important roles in cellular signaling, cell-cell interactions, and tissue homeostasis. Patients with / XDPKD type 2 (ADPKD2) have mutations in the PKD2 gene and make up around 15% of the total ADPKD patient population. Mutations in the PKD2 gene can cause the protein level of PC2 to drop below a “cystic threshold” in cells within the proximal tubule, distal tubule, and collecting duct regions of the kidney nephron, thus driving ADPKD2 pathogenesis. Gallagher et al., Adv Chronic Kidney Dis., 17(2): 118-30 (2010); Lakhia el al., Nat Commun. 13(1):4765 (2022).Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0006] Conventional treatment for ADPKD focuses on managing symptoms, controlling complications, and slowing the progression of kidney damage. Such methods are limited in efficacy because they do not directly target the underlying genetic cause of the disease.SUMMARY

[0007] The present disclosure relates to compositions and methods for the treatment of autosomal dominant polycystic kidney disease (ADPKD). In some embodiments, the present disclosure provides compositions and methods for the treatment of ADPKD type 2 (ADPKD2), which is caused by mutations in the PKD2 gene. In some embodiments, the present disclosure provides nucleic acid molecules comprising a polynucleotide encoding a polycystin-2 (PC2) for the treatment of ADPKD2

[0008] In some embodiments, the present disclosure provides a nucleic acid molecule comprising a polynucleotide encoding a PC2 protein, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 3- 5 and 7-9. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 5 or SEQ ID NO: 9.

[0009] In some embodiments, the present disclosure provides a vector comprising the nucleic acid molecule described herein. In some embodiments, the vector is a recombinant adeno-associated virus (rAAV) vector.

[0010] In some embodiments, the present disclosure provides an rAAV particle comprising: (a) the nucleic acid molecule or the vector described herein; and (b) a capsid of AAV1, AAV2, AAV2i8, AAV3, AAV3-B, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh8R, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAV-DJ, AAV LK03, AAV KP1, AAV.cc47, AAV.cc84, AAVrh74, AAV44-9, or a variant thereof. In some embodiments, the capsid protein is an AAV.kl3 capsid protein. In some embodiments, the capsid protein is an AAV.k20 capsid protein.

[0011] In some embodiments, the present disclosure provides a pharmaceutical composition, comprising any of the nucleic acid molecules, vectors, or rAAV particles disclosed herein, and a pharmaceutically acceptable excipient.7Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0012] In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of any of the nucleic acid molecules, vectors, rAAV particles, or pharmaceutical compositions disclosed herein. In some embodiments, the disease or disorder is ADPKD2.BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Fig. 1 shows a schematic of PKD2 cassette designs. These cassettes represent a combination of four different promoter elements (chicken beta actin [CBA], human phosphoglycerate kinase [hPGK], glucose-6-phosphatase [G6Pase], and mouse Pkd2 [mPKD2]) and four different PKD2 coding sequences (CDSs) (full-length, wild-type PKD2 CDS [PKD2-FL], a codon-optimized PKD2 CDS [PKD2-FL; CO], a CpG-depleted PKD2 CDS [PKD2-FL; ACpG], and a codon-optimized, CpG-depleted PKD2 CDS [PKD2-FL; CO; ACpG], Each cassette was synthesized with and without a C-terminal linker and a 3X hemagglutinin (HA) tag and contains a bovine growth hormone polyad enylati on signal (bGHpA). Each cassette is flanked by inverted terminal repeats (ITRs).

[0014] Figs. 2A-2C show that Ffa / 2-knockout mouse inner medullary collecting duct cells (IMCD3 cells) transfected with plasmids containing HA-tagged PKD2 cassettes from Fig. 1 expressed varying levels of PKD2 mRNA and PC2 protein. Fig. 2A shows the expression level of PKD2 mRNA following transfection of PAz / 2-knockout IMCD3 cells with either HA-tagged PKD2 or untagged PKD2 cassettes of Fig. 1. Fig. 2B shows the expression level of PKD2 mRNA normalized to the expression level of GAPDH mRNA following transfection of Av / 2-knockout IMCD3 cells with either HA-tagged PKD2 or untagged PKD2 cassettes of Fig. 1. In Fig. 2A and Fig. 2B, RNA expression was measured by reverse transcription followed by digital PCR (RT-dPCR; copies / ng). N:::3 for all samples. Fig. 2C shows the expression level of PC2 protein relative to the CBA-PKD2-FL cassette following transfection of F z / 2-knockout IMCD3 cells with the HA-tagged PKD2 cassettes of Fig. 1. PC2 expression was measured via the HA tag signal using capillary electrophoresis-based immunodetection. N:::3 for all samples.

[0015] Figs. 3A and 3B show that relative PC2 protein expression is affected by PKD2 coding sequences in a species / cell-dependent manner following transfection of plasmids containing HA- tagged PKD2 cassettes containing a CBA promoter. Fig. 3A shows the expression level of PC2 protein in HEK293T cells following transfection with HA-tagged PKD2 cassetes containing aAttorney Docket No. TORQ-012 / 02WO 339010-2071CBA promoter. Fig.3B shows the expression level of PC2 protein in FAz / 2-knockout IMCD3 cells following transfection with HA-tagged PKD2 cassettes containing a CBA promoter. In Fig. 3A and Fig. 3B, PC2 expression for all cassettes was measured via the HA tag signal using capillary electrophoresis-based immunodetection and plotted relative to the PKD2-FL cassette. NT, no treatment control; HA-GFP, plasmid encoding N-temiinal HA-tagged EGFP as transfection control. N=3 for all samples. Statistical analysis via one-way ANOVA with Dunnett’s multiple comparisons to PKD2-FL expression. ** p < 0.01, **** p < 0.0001.

[0016] Figs. 4A-4E show that FAzZ2-knockout IMCD3 cells transduced with AAV-packaged PKD2 cassettes at a multiplicity7of infection (MOI) of 5E6, 1E6, or 5E5 expressed varying levels of PKD2 mRNA and PC2 protein. Fig.4A shows the expression level of PKD2 mRNA following transduction with AAV-packaged PKD2 cassettes. Fig. 4B shows the expression level of PKD2 mRNA normalized to the expression level of GAPDH mRNA following transduction with AAV-packaged PKD2 cassettes. In Fig. 4A and Fig. 4B, RNA expression was measured by reverse transcription followed by digital PCR (RT-dPCR; copies / ng). N=3 for all samples. Figs. 4C and 4D show that transduced PAz / 2-knockout IMCD3 cells expressed varying levels of the PC2 protein. PC2 protein expression was measured via the HA tag signal using capillary electrophoresis-based immunodetection. N:::3 for all samples. Fig. 4C shows the fold change in PC2 protein levels in A / XV-packaged PKD2 cassettes transduced at an MOI of 5E6, 1E6, or 5e5 relative to the CBA-PKD2-FL cassette transduced at an MOI of 5e6. Fig. 4D is a representative image of PC2 protein expression from the treatment groups shown in Fig. 4C. Fig. 4E shows that, where PC2 protein expression was detectable, PKD2 mRNA expression strongly correlated with PC2 protein expression (R2= 0.8132) in 7W2-knockout IMCD3 cells transduced with AAV-packaged PKD2 cassettes from Fig. 4A and Fig.4C.

[0017] Figs.5A-5B show PC2 protein expression in Pfc / 2-knockout 1MCD3 cells transduced with AAV-packaged PKD2 cassettes at an MOI of 5E5, 1E6, or 5E6. Fig. 5A shows the percentage of PC2-expressing cells following transduction with AAV-packaged PKD2 cassettes. Fig. 5B shows the fold change in PC2 protein levels following transduction with AAV-packaged PKD2 cassettes relative to a no treatment control (NT). PC2 protein was measured via the HA tag signal using single cell immunocytochemistry. N=3 for all samples.

[0018] Fig.6A shows a schematic of PKD2 cassettes designed to modulate PC2 expression by: i) decreasing liver expression through addition of liver-specific miR-22 binding sites (left panel); ii)Attorney Docket No. TORQ-012 / 02WO 339010-2071using promoters with kidney-specific activity (middle panel); or iii) using truncated PKD2 promoters (right panel). PAS, polyadenylation signal. Figs. 6B and 6C show that PKD2 mRNA was expressed from the PKD2 cassettes described in Fig. 6A following transfection in mouse Pfc / 2-knockout IMCD3 cells (Fig. 6B) and human PKD2-knockout HEK293 cells (Fig. 6C). In Fig. 6B and Fig. 6C, the dotted lines represent mRNA copies from reference CBA (PTR089) and hPGK (PTR101) cassettes. Fig. 6D shows that PC2 protein was detectable in a subset of PKD2 cassettes described in Fig. 6A following transfection in PAD2-knockout HEK293 cells. ND, not detectable. The dotted lines represent PC2 signal from reference CBA (PTR089) and hPGK (PTR101) cassettes. Fig. 6E shows a positive correlation between PC2 protein and PKD2 mRNA in FKD -knockout HEK293 cells transfected with the PKD2 cassettes from Fig. 6A. The solid line represents a simple linear regression fit. Fig. 6F is a table listing the PKD2 cassettes in which PC2 protein was either detectable (dark font) or not detectable (light font) in PXD2-knockout HEK293 cells following transfection.

[0019] Fig. 7 shows a schematic of hPGK-dnven PKD2 cassettes with additional 5’ and 3’ untranslated region (UTR) elements, as listed in the tables. Not shown is a cassete incorporating five miR-122 binding sites downstream of the PKD2-FL; CO; ACpG coding sequence (PTR327). PAS, polyadenylation signal. Fig. 7B shows PKD2 mRNA in PKD2-knockout HEK293 cells transfected with the PKD2 cassettes shown m Fig. 7A. Fig. 7C shows PC2 protein in PKD2-knockout HEK293 cells transfected with the PKD2 cassettes shown in Fig. 7A. In Fig. 7B and Fig.7C, the dotted line represents mRN / X copies ( Fig. 7B) or PC2 signal (Fig. 7C) from a reference hPGK PKD2 cassette (PTR101 ). Kruskal- Wallis with Dunn’s multiple comparison test using nontransformed values was performed on the RNA dataset to compare the mean of each cassette to the mean of PTR101 (Fig. 7B), and one-way ANOVA with Tukey’s multiple comparison test using square root-transformed values was performed on the protein dataset to compare the mean of each cassette to the mean of PTR101 (Fig. 7C). For Fig. 7B, no significant differences were found for any of the comparisons. For Fig. 7C, PTR285 resulted in a significant increase in PC2 expression compared to PTR101 (****, p <0.0001). Fig. 7D shows average PC2 protein levels per average PKD2 mRNA levels in transfected PKD2-knockout HEK293 cells for three replicates based on the average data from Fig. 7B and Fig. 7C. Fig. 7E shows a positive correlation between PC2 protein and PKD2 mRNA in FAD2-knockout HEK293 cells transfected with the PKD2 cassettes from Fig. 7A. The solid line represents a simple linear regression fit.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0020] Fig.8 A shows a schematic of the study d esign for evaluation of AAV.k20-packaged PKD2 cassettes in naive C57B1 / 6 mice. Mice were dosed via tail vein injection at six weeks of age with vectors containing either a CpG-depleted or codon-optimized and CpG-depleted PKD2 transgene driven by either a CBA or hPGK promoter. Each vector was tested at doses of 1E14 and 3E14 vg / kg. The CBA-driven codon-optimized and CpG-depleted PKD2 vector was additionally tested at 8.6E14 vg / kg. A control group of mice was injected with formulation buffer (vehicle). Mice were euthanized 28 days post-injection at approximately 10-weeks of age. Figs. 8B and 8C show that AAV.k20 vector genomes and FAD2 mRNA were detected in kidney via dPCR or RT-dPCR, respectively, for all AAV,k20-packaged PKD2 cassettes evaluated. Copy numbers were normalized per 1000 ng gDNA input for vector genome analysis (Fig. 8B) or per 100 ng cDNA input for mRNA analysis (Fig. 8C), Mouse kidneys were sectioned, free-floating paraffin-embedded, and immunohistochemistry (IHC) stained for HA-tagged PC2 protein. Stained slides were imaged using a slide scanner, and images were analyzed via HALO quantitative image analysis software. Fig. 8D show's exemplary IHC images with PC2-HA protein expression from AAV076 in mouse kidney cortex and medullar tissue (arrows) and Fig. 8E quantifies the percentage of PC2-HA+ cells in the kidney following delivery of the AAV.k20-packaged PKD2 cassettes described in Fig. 8A. Mouse kidneys were sectioned, free-floating paraffin-embedded, and immunofluorescence stained for PC2-H / X, OAT-1, ECAD, and AQP2 using an automatic Stainer. Stained slides were imaged using a slide scanner, and images were analyzed via HALO quantitative image analysis software. Fig. 8F shows PC2-HA colocalization with cell type-specific markers for proximal tubule cells (OAT1+), distal tubule cells (ECAD+OAT1-), and collecting duct cells (AQP2-r). The arrow points to PC2-HA colocalization with the specific cell type of interest. Fig. 8G quantifies the percentage of HA+ cells in proximal tubules (PTs), distal tubules (DTs), and collecting ducts (CDs) following delivery of the AAV.k20-packaged PKD2 cassettes described in Fig. 8A.

[0021] Fig. 9A shows a schematic of the study design for evaluation of the AAV.k20-packaged PKD2 cassettes containing either the CBA promoter (AAV076) or hPGK promoter (AAV128) driving a codon-optimized and CpG-depleted PKD2 transgene in naive domestic farm pigs. Pigs were dosed unilaterally via retrograde ureteral administration (RUA) at 3E14 vg / kidney. Pigs were dosed at approximately three months of age and sacrificed 28 days following RUA. A control group of pigs was treated with formulation buffer. Figs. 9B-9C show’ that AAV.k20 vectorAttorney Docket No. TORQ-012 / 02WO 339010-2071genomes and PKD2 mRNA were detected m kidney via dPCR or RT-dPCR, respectively, for the AAV.k20-packaged PKD2 cassettes evaluated (AAV076 and AAV128). Copy numbers were normalized per 1000 ng gDNA input for vector genome analysis (Fig. 9B) or per 100 ng cDNA input for mRNA (Fig. 9C). The whole kidney is represented by 23 biopsies across the inner medulla, outer medulla, and cortex. The dotted lines represent the lower limits of quantification for vector biodistribution (Fig. 9B) and RNA expression (Fig. 9C) and are 250 copies / 1000 ng gDNA and 25 copies / 100 ng cDNA, respectively. Fig. 9D shows exemplary IHC images with PC2-HA expression (brown staining) in pig kidney cranial and caudal poles following delivery of AAV076. Fig. 9E shows a lack of PC2-HA expression in pigs administered formulation buffer (negative control). Fig. 9F shows the percentage of HA+ tissue area in whole sections of the pig kidney. Fig. 9G shows PC2-HA expression in proximal tubule (PT) cells, distal tubule (DT) cells, and collecting duct (CD) cells identified via cell morphology. In Figs. 9D-9G, pig kidneys were sectioned, free-floating paraffin-embedded, and immunohistochemistry-stained for HA-tagged PC2 protein. Stained slides were imaged using a slide scanner, and images were analyzed via ImageJ (Fig. 9F).

[0022] An A / XV.k20-packaged PKD2 cassette containing the CBA promoter driving a codon- optimized and CpG-depleted PKD2 transgene (AAV076) was evaluated in naive domestic farm pigs via bilateral RUA. The pigs were dosed with AAV076 at either 1E14, 3E14, or 1E15 vg / kidney at approximately three months of age and sacrificed 28 days following RUA. Figs. 10A and 10B show that? XAV.k20 vector genomes and PKD2 mRNA were detectable via dPCR or RT-dPCR, respectively, in the kidneys of pigs administered with AAV076 via RUA. Copy numbers were normalized per 1000 ng gDNA input for vector genome analysis (Fig. 10A) or per 100 ng cDNA input for mRNA (Fig. 10B). The whole kidney is represented by 23 biopsies across the inner medulla, outer medulla, and cortex. The dotted lines represent the lower limits of quantification for vector biodistribution and RNA expression and are 5000 copies / 1000 ng gDNA and 600 copies / 100 ng cDNA, respectively.

[0023] Figs. 10C-10F show that AAV.k20 vector genomes and PKD2 mRN A were detectable via dPCR or RT-dPCR, respectively, for most doses evaluated m the liver, heart, bladder, and ureter. Copy numbers were normalized per 1000 ng gDNA input for vector genome analysis (left panels of Figs. 10C-10F) or per 100 ng cDNA input for mRNA (right panels of Figs. 10C-10F). The whole liver is represented by four biopsies across left and right lateral and medial lobes (Fig. 10C)Attorney Docket No. TORQ-012 / 02WO 339010-2071and the whole heart is represented by four biopsies across the left and right atrium and left and right ventricle (Fig. 10D). The whole bladder and whole ureter are represented by two biopsies each (Figs. 10E-10F). In Fig. IOC, the dotted lines represent the lower limits of quantification for vector biodistribution and RNA expression and are 5000 copies / 1000 ng gDNA and 600 copies / 100 ng cDNA, respectively.

[0024] Fig. 10G shows exemplary IHC images with PC2-HA expression m kidney cranial and caudal poles of pigs administered AAV076 and a lack of PC2-HA expression in the kidney cranial poles of pigs administered formulation buffer. Fig. 10H shows the percentage of HA+ tissue area in whole sections of the pig kidney. The 1E15 vg / kidney dose resulted in significantly higher %HA positive area I section compared to other groups (one-way ANOVA w / Tukey’s multiple comparisons comparing all means, ** p < 0,01, *** p < 0.001). Pig kidneys were sectioned, treefloating paraffin-embedded, and IHC-stained for HA-tagged PC2 protein. Stained slides were imaged using a slide scanner, and images were analyzed via Image!

[0025] Fig. 11A shows a schematic of the study design for evaluation of the biological activity of AAV076 in an ADPKD2 disease mouse model. In this model, Pkd2 knockout in the kidney and the onset of cystic disease can be induced upon doxycycline administration. Mice contained a Pkd2a°Yj' genefwith exons 2-4 flanked by LoxP sites on one allele and a null allele with a targeted mutation in exon 1 leading to loss of expression, a Pax8rtTAallele, and a Tet()Creallele. This genotype is generated by crossing the Pkd2'" mouse strain with the Pkd2fiox / flox, Pax8rtTA, TetOcremouse strain. Both strains were supplied by Yale University. Upon administration of doxycycline, Cre recombinase was expressed specifically in renal epithelial cells, leading to knockout of endogenous Pkd2 expression. Mice were dosed with AAV076 at post-natal day 1 (Pl) via temporal facial vein injection at a dose of IE 14 vg / kg. Mice were then administered doxycycline at 50 mg / kg body weight for three consecutive days from Pl 1 -13, and mice were euthanized at P28 for analyses. No disease induction control mice (-Dox) were not treated with doxycycline or AAV. Disease-only control mice (+Dox) were treated with doxycycline but not AAV. Fig. 11B shows the kidney weight to body weight (KW / BW) ratio, which was the sum weight of both kidneys normalized to the total body weight. Fig. 11C shows AAV076 vector genomes in the kidneys of untreated (-Dox) mice or mice treated with doxycycline only (+Dox) or doxycycline and AAV076 (+Dox, AAV076). AAV076 vector genomes were determined by dPCRand copy numbers were normalized per 1000 ng gDNA input. Fig. 11D shows PKD2 mRNA expression in the kidneys of untreated (-Dox) miceAttorney Docket No. TORQ-012 / 02WO 339010-2071or mice treated with doxycycline only (TDox) or doxycycline and AAV076 (TDox, AAV076). PKD2 mRNA expression was determined by RT-dPCR and normalized per 100 ng cDNA input. In Figs. 11B-11D, statistical significance was determined via a one-way ANOVA with Holm-Sidak’s multiple comparisons test between the means of the TDox and AAV076 TDox treatment groups using loglO-transformed values (* p < 0.05, **** p < 0.0001).DETAILED DESCRIPTION

[0026] The present disclosure provides nucleic acid molecules designed to supplement the expression of polycystin-2 (PC2) in autosomal dominant polycystic kidney disease (ADPKD) subjects with mutations in the PKD2 gene. Expressing such nucleic acid molecules in cells with mutations in the PKD2 gene restores PC2 protein expression to above the cystic threshold, thus preventing the cells from entering a hyperproliferative state and slowing the rate of new cyst formation. Contrary to conventional methods of treating ADPKD, methods provided herein address the underlying genetic cause of ADPKD.Definitions

[0027] Unless otherwise defined herein, technical and scientific terms used in the present description have the meanings that are commonly understood by those of ordinary skill in the art. For purposes of interpreting this specification, the following description of terms will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa unless the content clearly dictates otherwise. In the event that any description of a term set forth conflicts with any document incorporated herein by reference, the description of the term set forth below shall control.

[0028] The terms “a”, “an”, and “the”, as used herein, include plural references unless the context clearly dictates otherwise. As such, the terms “a,” “an,” “one or more,” and “at least one” are used interchangeably herein. In addition, reference to “an element” by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there is one and only one of the elements.

[0029] Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the device or the method being employed to determine the value, or the variation that exists among the samples being measured. Unless otherwise stated or otherwiseAttorney Docket No. TORQ-012 / 02WO 339010-2071evident from the context, the term “about” means within 10% above or below the reported numerical value (except where such number would exceed 100% of a possible value or go below 0%). When used in conjunction with a range or series of values, the term “about” applies to the endpoints of the range or each of the values enumerated in the senes, unless otherwise indicated. As used in this application, the terms “about” and “approximately” are used as equivalents.

[0030] The term “and / or”, as used herein, refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”).

[0031] The term “between”, as used in a phrase as such “between A and B” or “between A-B” refers to a range including both A and B.

[0032] The terms “comprise” and its grammatical equivalents, as used herein, specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.

[0033] The terms “including”, “includes”, “included”, and other forms, as used herein, are not limiting.

[0034] The term “polynucleotide” or “nucleic acid”, as used herein, refers to deoxyribonucleic acid (DNA), ribonucleic acid (RNA) and DNA / RNA hybrids. Polynucleotides may be single¬ stranded or double-stranded and either recombinant, synthetic, or isolated. Polynucleotides include, but are not limited to: pre-messenger RNA (pre-mRNA), messenger RNA (rnRNA), RNA, genomic DNA (gDNA), PCR amplified DNA, complementary DNA (cDN A), synthetic DNA, or recombinant DNA. Polynucleotides can comprise modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or R A polymerase or by a synthetic reaction. Unless specified otherwise, the left-hand end of any single-stranded polynucleotide sequence disclosed herein is the 5’ end; the left-hand direction of double-stranded polynucleotide sequences is referred to as the 5’ direction. The direction of 5’ to 3’ addition of nascent RNA transcripts is referred to as the transcription direction.

[0035] The term “coding sequence” or a polynucleotide which “encodes” a polypeptide, as used herein, is a nucleic acid molecule which is transcribed (in the case of DNA) and translated (in the case of rnRNA) into a polypeptide when placed under the control of appropriate regulatory' sequences. The boundaries of the coding sequence are determined by a start codon at the 5’ (amino)Attorney Docket No. TORQ-012 / 02WO 339010-2071terminus and a translation stop codon at the 3’ (carboxy) terminus. A transcription termination sequence may be located 3’ to the coding sequence. A Kozak sequence may be located 5’ to the coding sequence.

[0036] The term “operatively linked” and similar phrases (e.g., operably linked, genetically fused), as used herein, refer to the operational linkage of nucleic acid sequences or amino acid sequences placed in functional relationships with each other. For example, a promoter operatively linked to a polynucleotide encoding a polypeptide result in the transcription of the polynucleotide and ultimately the expression of the polypeptide. As another example, an operatively linked peptide is one in which the functional domains are placed with appropriate distance from each other to impart the intended function of each domain.

[0037] As used herein, the term “promoter” refers to a DNA regulatory region capable of binding RNA polymerase and initiating transcription of a downstream coding or non-coding sequence.

[0038] The terms “polypeptide” and “peptide” and “protein”, as used herein, refer to polymers of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification. Also included within the definition are, for example, polypeptides containing one or more analogs of an ammo acid, including but not limited to, unnatural ammo acids, as well as other modifications known in the art.

[0039] The term “amino acid”, as used herein, refers to any naturally occurring amino acid or modified forms thereof, non-naturally occurring amino acids, and synthetic ammo acids. In some embodiments, the ammo acid is modified by post-translational modification (e.g., acetylation, amidation, formylation, hydroxylation, methylation, phosphorylation, or sulfatation). Examples of modified amino acids include, but are not limited to, 2-amidoadipic acid, 3-ammoadipic acid, betaalanine, beta-aminoproprionic acid, 2-aminobutyric acid, 4-aminobutyric acid, piperidinic acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, t-butylalanine, citrulline, cyclohexylalanine, 2,4-diaminobutyric acid, desmosine, 2,21-diaminopimelic acid, 2,3-diaminoproprionic acid, N-etliylglycine, N-ethylasparagme, homoarginine, homocysteine, homoserine, hydroxylysine, allo-hydroxylycme, 3- hydroxyroline, 4-hydroxyproline, isodesmosine, allo-isoleucine, methionine sulfoxide, N-Attorney Docket No. TORQ-012 / 02WO 339010-2071methylglycine, sarcosine, 6-N-methyllycine, N-methylvaline, 2-naphthylalanine, norvaline, norleucine, ornithine, 4-chlorophenylalanine, 2-fluorophenylalanine, 3 -fluorophenylalanine, 4-fluorophenylalanine, phenylglycine, and beta-2-thienylalanine. In some embodiments, the amino acid is a non-naturally occurring ammo acid or “unnatural” amino acid as described by Wang et al., Annu Rev Biophys Biomol Struct, 35:225-49 (2006).

[0040] The term “synthetic”, as applied to a nucleic acid, a polypeptide, a cell, or an organism, refers to a nucleic acid, polypeptide, cell, or organism that cannot be directly isolated from a source in nature. In some embodiments, the synthetic nucleic acid, polypeptide, cell, or organism is substantially similar as compared to the corresponding natural -occurring one. In some embodiments, the synthetic nucleic acid, polypeptide, cell, or organism is altered or changed as compared to the corresponding natural-occurring one. In some embodiments, the synthetic nucleic acid, polypeptide, cell, or organism is produced by functionally linking or combining different fragments of sourcing nucleic acids, polypeptides, cells, or organisms together. For instance, a synthetic polypeptide can comprise different sourcing polypeptides functionally linked together.

[0041] The term “vector”, as used herein, refers to a molecule or moiety which transports, transduces, or transfects a nucleic acid molecule of interest into a host cell or tissue. Vectors applicable for use include, for example, plasmids, phage vectors, viral vectors, episomes, and artificial chromosomes. A vector may include sequences that direct autonomous replication in a cell, or may include sequences sufficient to allow integration into host cell DNA. Additionally, the vectors can include one or more selectable marker genes and appropriate expression control sequences. Selectable marker genes that can be included, for example, provide resistance to antibiotics or toxins, complement auxotrophic deficiencies, or supply critical nutrients not in the culture media. Expression control sequences can include constitutive and inducible promoters, transcription enhancers, transcription terminators, and the like, which are well known in the art. The introduction of nucleic acid molecules into a host cell can be confirmed using methods well known in the art. Such methods include, for example, nucleic acid analysis such as Northern blots or polymerase chain reaction (PCR) amplification of mRNA, immunoblottmg for expression of gene products, or other suitable analytical methods to test the expression of an introduced nucleic acid sequence or its corresponding gene product. It is understood by those skilled in the art that the nucleic acid molecules are expressed in a sufficient amount to produce a desired product andAttorney Docket No. TORQ-012 / 02WO 339010-2071it is further understood that expression levels can be optimized to obtain sufficient expression using methods well known in the art.

[0042] As used herein, the term “sequence identity” refers to the extent to which two optimally aligned polynucleotides or polypeptide sequences are invariant throughout a window of alignment of residues, e.g., nucleotides or amino acids. An “identity fraction” for aligned segments of a test sequence and a reference sequence is the number of identical residues which are shared by the two aligned sequences divided by the total number of residues in the reference sequence segment, i.e., the entire reference sequence or a smaller defined part of the reference sequence. “Percent identity” is the identity fraction times 100. Comparison of sequences to determine percent identity can be accomplished by a number of well-known methods, including for example by using mathematical algorithms, such as, for exampl e, those in the BLAST suite of sequence analysis programs. Unless noted otherwise, the term “sequence identity” in the claims refers to sequence identity as calculated by Clustal Omega® using default parameters.

[0043] The term “flanked”, as used herein, with respect to a sequence that is flanked by other elements, indicates the presence of one or more the flanking elements upstream and / or downstream, i.e., 5’ and / or 3 relative to the sequence. The term “flanked” is not intended to indicate that the sequences are necessarily contiguous. For example, there may be intervening sequences between the nucleic acid encoding the gene of interest and a flanking element. A nucleic acid molecule that is “flanked” by two other elements indicates that one element is located 5’ to the sequence and the other is located 3’ to the sequence; however, there may be intervening sequences therebetween.

[0044] The term “inverted terminal repeat” or “HR” sequence, as used herein, refers to relatively short sequences found at the termini of viral genomes which are in opposite orientation. An “AAV inverted terminal repeat (ITR)” sequence is well known in the art, and is usually an approximately 145 -nucleotide sequence that is present at both termini of the native single-stranded AAV genome. The outermost 125 nucleotides of the ITR can be present in either of two alternative orientations, leading to heterogeneity between different AAV genomes and between the two ends of a single AAV genome. The outermost 125 nucleotides also contains several shorter regions of selfcomplementarity (designated A, A’, B, B’, C, C’ and D regions), allowing intra-strand base-pairing to occur within this portion of the ITR.

[0045] The term “recombinant” means a genetic entity distinct from that generally found in nature. As applied to a polynucleotide or gene, this means that the polynucleotide is the product of variousAttorney Docket No. TORQ-012 / 02WO 339010-2071combinations of cloning, restriction and / or ligation steps, and other procedures that result in the production of a construct that is distinct from a polynucleotide found in nature.

[0046] The term “recombinant AAV vector (rAAV vector)”, as used herein, refers to a polynucleotide vector comprising a nucleic acid sequence from an AAV and one or more heterologous sequences (i.e., nucleic acid sequence not of AAV origin). In some embodiments, the one or more heterologous sequences are flanked by at least one, In some embodiments two, AAV inverted terminal repeat sequences (ITRs). In some embodiments, such rAAV vectors can be replicated and packaged into infectious viral capsid particles, e.g., when present in a host cell that has been infected with a suitable helper virus (or that is expressing suitable helper functions) and that is expressing AAV rep and cap gene products (i.e., A AV Rep and Cap proteins). An rAAV vector may be incorporated into a larger polynucleotide (e.g,, in a chromosome or in another vector such as a plasmid used for cloning or transfection), and can be “rescued” by replication and encapsidation in the presence of AAV packaging functions and suitable helper functions. An rAAV vector can be in any of a number of forms, including, but not limited to, plasmids, linear artificial chromosomes, complexed with lipids, encapsulated within liposomes, and encapsulated in a viral capsid particle, particularly an AAV particle. / Xn rAAV vector can be packaged into an AAV capsid to generate a “recombinant adeno-associated viral capsid particle (rAAV particle).”

[0047] The term “capsid”, as used herein, refers to a capsid protein of a virus, such as an adeno- associated virus, wherein the capsid encapsulates a nucleic acid molecule or viral genome. In some embodiments, the capsid is a variant capsid. The term “variant capsid” refers to a capsid protein that has been modified (e.g., one or more ammo acid substitutions) compared to a parental capsid protein. In some embodiments, the variant capsid protein comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the native or parental capsid protein. The term “AAV capsid” or “AAV capsid protein” or “AAV cap”, as used herein, refers to a protein encoded by an AAV capsid (cap) gene (e.g., VPI, VP2, and VP3) or a variant thereof. For example, the term includes but not limited to a capsid protein derived from any AAV serotype such as AAV1, AAV2, AAV2i8, AAV3, AAV3-B, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh8R, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAV-DJ, AAV-2 / 1, AAV 2 / 6, AAV 2 / 7, AAV 2 / 8, AAV 2 / 9, AAV LK03, AAVrhlO, AAVrh74, AAV44-9, or a variant thereof. In someAttorney Docket No. TORQ-012 / 02WO 339010-2071embodiments, the capsid is an AAV9 capsid variant, such as AAV.kl3 or AAV.k20. The term also includes a capsid protein expressed by or derived from a recombinant AAV such as a chimeric AAV. The term “AAV capsid particle” or “AAV particle”, as used herein, includes at least one AAV capsid protein (e.g., a VP1 protein, a VP2 protein, a VP3 protein, or variant thereof) and optionally encapsulates a nucleic acid from an AAV genome or a nucleic acid derived from an AAV genome. The term “serotype” used with respect to vector or virus capsid is defined by a distinct immunological profile based on the capsid protein sequences and capsid structure.

[0048] The term “transduced”, as used herein, refers to a process by which a transgene is introduced into a host cell from a virus particle.

[0049] The term “tropism”, as used herein, refers to preferential erury of a virus into a certain cell or tissue, optionally followed by expression of nucleic acid sequences carried by the viral genome in the cell or tissue.

[0050] As used herein, the term “subject” refers to any subject, e.g., a human or a non-human mammal, for whom diagnosis, prognosis, or therapy is desired. The term “subject” may mean a human or non-human mammal affected, likely to be affected, or suspected to be affected with a disease. The terms “subject” and “patient” are used interchangeably herein. In some embodiments, a subject is a mammal. A mammal includes primates, such as humans, monkeys, chimpanzee, and apes, and non-primates such as domestic animals, including laboratory animals (such as rabbits and rodents, e.g., guinea pig, rat, or mouse) and household pets and farm animals (e.g., cats, dogs, swine, cattle, sheep, goats, horses, rabbits), and non-domestic animals, such as wildlife, birds, reptile, fish, or the like. As used herein, the term “a subject in need thereof” includes subjects that could or would benefit from the methods described herein. Subjects in need of treatment include, without limitation, those already with the disease, disorder, or condition, those prone to having the disease, disorder, or condition, those in which the disease, disorder, or condition is suspected, as well as those in which the disease, disorder, or condition is to be prevented, ameliorated, or reversed.

[0051] The term “pharmaceutical composition” or “therapeutic composition”, as used herein, refers to a composition capable of being administered to a subject for the treatment of a particular disease or disorder, such as ADPKD.

[0052] The term “pharmaceutically acceptable excipient, carrier or diluent”, as used herein, refers to any substance formulated alongside the active ingredient of a pharmaceutical composition thatAttorney Docket No. TORQ-012 / 02WO 339010-2071allows the active ingredient to retain biological activity and is non-reactive with the subject’s immune system. Such a substance can be included for the purpose of long-term stabilization, bulking up solid formulations that contain potent active ingredients in small amounts, or to confer a therapeutic enhancement on the active ingredient in the final dosage form, such as facilitating absorption, reducing viscosity, or enhancing solubility. The selection of appropriate substance can depend upon the route of administration and the dosage form, as well as the active ingredient and other factors. Compositions having such substances can be formulated by well-known conventional methods (see, e.g., Remington, The Science and Practice of Pharmacy, 23rd edition, A. Adejare, ed., Academic Press, 2020).

[0053] The term “administer”, “administration”, or “administering”, as used herein refers to the act of injecting or otherwise physically delivering a substance (e.g., a pharmaceutical composition provided herein) to a subject (e.g., human), such as by oral, mucosal, topical, intradermal, parenteral, intravenous, retrograde ureteral, intravitreal, intraarticular, subretmal, intramuscular, intrathecal delivery and / or any other method of physical delivery described herein or known in the art. The delivery can be systemic or to a specific tissue. In some embodiments, the pharmaceutical composition is administered by intravenous infusion. In some embodiments, the pharmaceutical composition is administered by retrograde ureteral infusion. In some embodiments, the pharmaceutical composition is administered by using an intravenous catheter (e.g., by inserting an IV catheter into a suitable peripheral vein for intravenous infusion of the pharmaceutical composition).

[0054] As used herein, the terms “prevent,” “preventing,” “prevention” and grammatical variants thereof refer to an approach for preventing the development of, or altering the pathology of, a condition, disease, or disorder. Accordingly, “prevention” may refer to prophylactic or preventive measures. In some embodiments, beneficial or desired clinical results include, but are not limited to, prevention or slowing of symptoms, progression or development of a disease, whether detectable or undetectable. A subject (e.g., a human) in need of prevention may thus be a subject not yet afflicted with the disease or disorder in question. The term “prevention” includes slowing the onset of disease relative to the absence of treatment and is not necessarily meant to imply permanent prevention of the relevant disease, disorder or condition. Thus “preventing” or “prevention” of a condition may in certain contexts refer to reducing the risk of developing the condition or preventing or delaying the development of symptoms associated with the condition.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0055] In some embodiments, the term “treating” refers to the treatment of a disease in a mammal, e.g., in a human, including (a) inhibiting the disease, i.e., arresting disease development or preventing disease progression; (b) relieving the disease, i.e., causing regression of the disease state or relieving one or more symptoms of the disease; and (c) curing the disease, i.e., remission of one or more disease symptoms. In some embodiments, treatment results in an improvement or remediation of the symptoms of the disease. In some embodiments, treatment refers to a short¬ term (e.g., temporary and / or acute) and / or a long-term (e.g., sustained) improvement or remediation in one or more disease symptoms. In some embodiments, the improvement is an observable or measurable improvement. In some embodiments, the improvement is an improvement in the general feeling of well-being of the subject,

[0056] As used herein, an “effective amount” or “therapeutically effective amount” is an amount or dose of a composition (e.g,, a therapeutic composition, compound, or agent) that produces at least one desired therapeutic effect m a subject, such as preventing or treating a target condition or beneficially alleviating a symptom associated with the condition. The most desirable therapeutically effective amount is an amount that will produce a desired efficacy of a particular treatment selected by one of skill in the art for a given subject in need thereof. This amount will vary depending upon a variety of factors understood by the skilled worker, including but not limited to the characteristics of the therapeutic composition (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (including age, sex, disease type, disease stage, general physical condition, responsiveness to a given dosage, and type of medication), the nature of the pharmaceutically acceptable carrier or carriers in the formulation, and the route of administration. One skilled in the clinical and pharmacological arts will be able to determine a therapeutically effective amount through routine experimentation, namely by monitoring a subject’s response to administration of a composition and adjusting the dosage accordingly (see e.g., Remington: The Science and Practice of Pharmacy (Gennaro A, ed., Mack Publishing Co., Easton, PA, U. S., 19th ed., 1995)).

[0057] General methods in molecular and cellular biochemistry can be found in such standard textbooks as Molecular Cloning: A Laboratory Manual, 3rd Ed. (Sambrook et al., HaRBor Laboratory Press 2001 ); Short Protocols in Molecular Biology, 4th Ed. (Ausubel et al. eds., John Wiley & Sons 1999); Protein Methods (Bollag et l., John Wiley & Sons 1996); Nonviral Vectors for Gene Therapy (Wagner et al. eds., Academic Press 1999); Viral Vectors (Kaplift & LoewyAttorney Docket No. TORQ-012 / 02WO 339010-2071eds., Academic Press 1995); Immunology Methods Manual (I. Lefkovits ed., Academic Press 1997); and Cell and Tissue Culture: Laboratory’ Procedures in Biotechnology (Doyle & Griffiths, John Wiley & Sons 1998), the disclosures of which are incorporated herein by reference.Nucleic Acid Molecules

[0058] The present disclosure provides nucleic acid molecules comprising a polynucleotide encoding a polycystin-2 (PC2) protein.

[0059] PC2 is encoded by the PKD2 gene (NCBI Gene ID: 5311; UmProtlD: Q13563). PC2 is a multifunctional, multi-pass transmembrane protein that plays important roles in cellular signaling, cell-cell interactions, and tissue homeostasis. PC2 functions as a calcium permeable cation channel and is involved in calcium transport and calcium signaling in renal epithelial cells. PC2 interacts with polycystin- 1 (PCI), and they may be partners in a common signaling cascade involved in tubular morphogenesis. Mutations in the PKD2 gene are associated with autosomal dominant polycystic kidney disease type 2 (ADPKD2).

[0060] In some embodiments, the PC2 protein comprises or consists of an amino acid sequence of SEQ ID NO: 1. In some embodiments, the PC2 protein comprises an amino acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99,5% sequence identity to an amino acid sequence of SEQ ID NO: 1, In some embodiments, the PC2 protein comprises an amino acid sequence of SEQ ID NO: 1 with one or more mutations therein. For example, in some embodiments, the PC2 protein comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 1. In some embodiments, the PC2 protein comprises an amino acid sequence of SEQ ID NO: 1 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0061] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence selected from any one of SEQ ID NOs: 2-11. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence selected from any one of SEQ ID NOs: 2-11. In some embodiments, the polynucleotide comprises a nucleic acid sequenceAttorney Docket No. TORQ-012 / 02WO 339010-2071selected from any one of SEQ ID NOs: 2-11 with one or more mutations therein. For example, in some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in any one of SEQ ID NOs: 2-11. In some embodiments, the polynucleotide comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 2-11 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0062] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence of SEQ ID NO: 2. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 2, In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 2 with one or more mutations therein. For example, in some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 2. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 2 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0063] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence of SEQ ID NO: 3. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 3. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 3 with one or more mutations therein. For example, m some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 3. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 3 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0064] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acidAttorney Docket No. TORQ-012 / 02WO 339010-2071sequence of SEQ ID NO: 4. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 4. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 4 with one or more mutations therein. For example, m some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 4. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 4 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0065] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence of SEQ ID NO: 5. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 5. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 5 with one or more mutations therein. For example, in some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 5. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 5 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0066] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence of SEQ ID NO: 6. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 6. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 6 with one or more mutations therein. For example, in some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9,Attorney Docket No. TORQ-012 / 02WO 339010-207110, or more mutations in SEQ ID NO: 6. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 6 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0067] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence of SEQ ID NO: 7. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 7, In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 7 with one or more mutations therein. For example, in some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 7. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 7 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations,

[0068] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence of SEQ ID NO: 8. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 8. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 8 with one or more mutations therein. For example, m some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 8. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 8 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0069] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises or consists of a nucleic acid sequence of SEQ ID NO: 9. In some embodiments, the polynucleotide comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, atAttorney Docket No. TORQ-012 / 02WO 339010-2071least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 9. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 9 with one or more mutations therein. For example, in some embodiments, the polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 9. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 9 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0070] The nucleic acid molecules encoding the PC2 protein disclosed herein may be combined with other DNA sequences, such as promoters and / or enhancers, untranslated regions (UTRs), signal sequences, Kozak sequences, polyadenylation signals, additional restriction enzyme sites, multiple cloning sites, internal ribosomal entry sites (IRES), recombinase recognition sites (e.g,, ITR, LoxP, FRT, and Att sites), termination codons, transcriptional termination signals, and polynucleotides encoding self-cleaving polypeptides, epitope tags, as disclosed elsewhere or as known in the art.

[0071] In some embodiments, the nucleic acid molecule encoding the PC2 protein is operably linked to a promoter. Varieties of promoters have been explored for gene expression in mammalian cells, and any of the promoters known in the art may be used in the present disclosure. Promoters may be roughly categorized as constitutive promoters or regulated promoters, such as inducible promoters.

[0072] In some embodiments, the nucleic acid molecule encoding the PC2 protein is operably linked to a constitutive promoter. Constitutive promoters allow heterologous genes (also referred to as transgenes) to be expressed constitutively in the host cells.

[0073] In some embodiments, the nucleic acid molecule encoding the PC2 protein is operably linked to an inducible promoter. Inducible promoters belong to the category of regulated promoters. The inducible promoter can be induced by one or more conditions, such as a physical condition, microenvironment of the engineered immune effector cell, or the physiological state of the engineered immune effector cell, an inducer (i.e., an inducing agent), or a combination thereof. In some embodiments, the inducing condition does not induce the expression of endogenous genes in the engineered mammalian cell, and / or in the subject that receives the pharmaceutical composition. In some embodiments, the inducing condition is selected from the group consistingAttorney Docket No. TORQ-012 / 02WO 339010-2071of: inducer, irradiation (such as ionizing radiation, light), temperature (such as heat), redox state, tumor environment, and the activation state of the engineered mammalian cell.

[0074] A person skilled in the art may recognize that a target cell may require a specific promoter including but not limited to a promoter that is species specific, inducible, tissue-specific, or cell cycle-specific Parr et al., Nat. Med. 3: 1145-9 (1997); the contents of which are herein incorporated by reference in its entirety. In some embodiments, the nucleic acid molecule comprises a promoter that is deemed to be efficient to drive the expression of the nucleic acid molecules described herein. Promoters that induce expression in most tissues include, for example, but are not limited to, human elongation factor la-subunit (EFla), immediate-early cytomegalovirus (CMV), the RSV LTR, the MoMLV LTR, the phosphoglycerate kinase- 1 (PGK) promoter, a simian virus 40 (SV 40) promoter and a CK6 promoter, a transthyretin promoter (TTR), a TK promoter, a tetracycline responsive promoter (TRE), an HBV promoter, an hAAT promoter, a LSP promoter, chimeric liver-specific promoters (LSPs), the telomerase (hTERT) promoter, chicken p-actm (CBA) and its derivative CAG and mmiCBA, the P glucuronidase (GUSB), or ubiquitin C (UBC). Tissuespecific expression elements can be used to restrict expression to certain cell types (e.g., kidney cells). The above-mentioned promoters can be combined with other synthetic short regulatory elements to generate novel synthetic promoters with center homology in DNA sequences.

[0075] In some embodiments, the nucleic acid molecule comprises a promoter. In some embodiments, the promoter is a chicken beta actin (CBA) promoter. In some embodiments, the promoter is a human phosphoglycerate kinase (hPGK) promoter. In some embodiments, the promoter is a human glucose-6-phosphatase (hG6Pase) promoter. In some embodiments, the promoter is a mouse polycystic kidney disease 2 (mPKD2) promoter.

[0076] In some embodiments, the nucleic acid molecule comprises a ubiquitous promoter. In some embodiments, the promoter is a chicken beta actin (CBA) promoter. In some embodiments, the promoter is a human phosphoglycerate kinase (hPGK) promoter. In some embodiments, the promoter is a human elongation factor la-subunit (EFla) promoter.

[0077] In some embodiments, the nucleic acid molecule comprises a promoter that is active in kidney tissue or kidney cells. In some embodiments, the promoter is a mouse E-cadherin promoter (mECAD). In some embodiments, the promoter is a mouse kidney-specific cadherin (mKSPC) promoter. In some embodiments, the promoter is a mouse paired box gene 8 (mPax8) promoter. In some embodiments, the promoter is a human with-no-lysine kinase 1 promoter with an upstreamAttorney Docket No. TORQ-012 / 02WO 339010-2071renal enhancer element (hWNKl-RP). In some embodiments, the promoter is a human phosphoglycerate kinase promoter with an upstream renal enhancer element (RP-hPGK).

[0078] In some embodiments, the nucleic acid molecule comprises a truncated PKD2 promoter (i.e., a mini-PKD2 promoter). In some embodiments, the truncated PKD2 promoter is a human truncated PKD2 promoter. In some embodiments, the truncated PKD2 promoter is a pig truncated PKD2 promoter. In some embodiments, the truncated PKD2 promoter is a mouse truncated PKD2 promoter.

[0079] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a promoter. In some embodiments, the promoter comprises or consists of a nucleic acid sequence selected from any one of SEQ ID NOs: 12-15 and 21-32, In some embodiments, the promoter comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence selected from any one of SEQ ID NOs: 12-15 and 21 -32. In some embodiments, the promoter comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 12-15 and 21-32 with one or more mutations therein. For example, in some embodiments, the promoter comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in any one of SEQ ID NOs: 12-15 and 21-32. In some embodiments, the promoter comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 12-15 and 21-32 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0080] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a promoter. In some embodiments, the promoter comprises or consists of a nucleic acid sequence of SEQ ID NO: 12. In some embodiments, the promoter comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 12. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 12 with one or more mutations therein. For example, in some embodiments, the promoter comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 12. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 12 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0081] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a promoter. In some embodiments, the promoter comprises or consists of a nucleic acid sequence of SEQ ID NO: 13. In some embodiments, the promoter comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 13. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 13 with one or more mutations therein. For example, in some embodiments, the promoter comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 13. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 13 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0082] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a promoter. In some embodiments, the promoter comprises or consists of a nucleic acid sequence of SEQ ID NO: 21, In some embodiments, the promoter comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 21. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 21 with one or more mutations therein. For example, in some embodiments, the promoter comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 21. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 21 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0083] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a promoter. In some embodiments, the promoter comprises or consists of a nucleic acid sequence of SEQ ID NO: 25. In some embodiments, the promoter comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 25. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 25 with one or more mutations therein. For example, in some embodiments, the promoter comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQAttorney Docket No. TORQ-012 / 02WO 339010-2071ID NO: 25. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 25 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0084] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a promoter. In some embodiments, the promoter comprises or consists of a nucleic acid sequence of SEQ ID NO: 28. In some embodiments, the promoter comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 28, In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 28 with one or more mutations therein. For example, in some embodiments, the promoter comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 28. In some embodiments, the promoter comprises a nucleic acid sequence of SEQ ID NO: 28 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0085] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a promoter. In some embodiments, the promoter comprises a renal enhancer element. In some embodiments, the renal enhancer element comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 161. In some embodiments, the renal enhancer element comprises a nucleic acid sequence of SEQ ID NO: 161 with one or more mutations therein. For example, in some embodiments, the renal enhancer element comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 161. In some embodiments, the renal enhancer element comprises a nucleic acid sequence of SEQ ID NO: 161 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0086] In some embodiments, the polynucleotide encoding the PC2 protein is operably linked to a polyadenylation (poly A) signal. The term “poly A signal” or “poly A sequence” as used herein denotes a DNA sequence which directs both the termination and polyadenylation of the nascent RNA transcript by RNA polymerase II. Poly A signals can promote mRNA stability by addition of a poly A tail to the 3 ’ end of the coding sequence and thus, contribute to increased translational efficiency. In some embodiments, the poly A sequence is an ideal poly A sequence (e.g., AATAAA,Attorney Docket No. TORQ-012 / 02WO 339010-2071ATT AAA, AGTAAA). In some embodiments, the poly A sequence is an SV40 poly A sequence, a bovine growth hormone poly A sequence (bGHpA), a rabbit P-globin poly A sequence (rPgpA), variants thereof, or another suitable heterologous or endogenous poly A sequence known in the art. In some embodiments, the polyad enylati on signal is a bovine growth hormone poly A (bGHpA) signal.

[0087] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a terminator. In some embodiments, the terminator comprises or consists of a nucleic acid sequence of SEQ ID NO: 18. In some embodiments, the terminator comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 18. In some embodiments, the terminator comprises a nucleic acid sequence of SEQ ID NO: 18 with one or more mutations therein. For example, in some embodiments, the terminator comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 18, In some embodiments, the terminator comprises a nucleic acid sequence of SEQ ID NO: 18 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0088] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a terminator. In some embodiments, the terminator comprises or consists of a nucleic acid sequence of SEQ ID NO: 33. In some embodiments, the terminator comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 33. In some embodiments, the terminator comprises a nucleic acid sequence of SEQ ID NO: 33 with one or more mutations therein. For example, in some embodiments, the terminator comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 33. In some embodiments, the terminator comprises a nucleic acid sequence of SEQ ID NO: 33 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0089] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises one or more untranslated regions. In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 5’ UTR. In some embodiments, the nucleic acid molecule encoding PC2 proteinAttorney Docket No. TORQ-012 / 02WO 339010-2071comprises a 3’ UTR. In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 5’ UTR and a 3’ UTR.

[0090] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 5’ UTR. In some embodiments, the 5’ UTR comprises or consists of a nucleic acid sequence selected from any one of SEQ ID NOs: 34-49. In some embodiments, the 5’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence selected from any one of SEQ ID NOs: 34-49, In some embodiments, the 5’ UTR comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 34-49 with one or more mutations therein. For example, in some embodiments, the 5’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in any one of SEQ ID NOs: 34-49. In some embodiments, the 5’ UTR comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 34-49 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0091] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 5’ UTR. In some embodiments, the 5’ UTR comprises or consists of a nucleic acid sequence of SEQ ID NO: 35. In some embodiments, the 5’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 35. In some embodiments, the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 35 with one or more mutations therein. For example, in some embodiments, the 5’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 35. In some embodiments, the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 35 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0092] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 3’ U TR. In some embodiments, the 3’ UTR comprises or consists of a nucleic acid sequence selected from any one of SEQ ID NOs: 50-61. In some embodiments, the 3’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5%Attorney Docket No. TORQ-012 / 02WO 339010-2071sequence identity to a nucleic acid sequence selected from any one of SEQ ID NOs: 50-61. In some embodiments, the 3’ UTR comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 50-61 with one or more mutations therein. For example, in some embodiments, the 3’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in any one of SEQ ID NOs: 50-61. In some embodiments, the 3’ UTR comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 50-61 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0093] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 3’ UTR. In some embodiments, the 3’ UTR comprises or consists of a nucleic acid sequence of SEQ ID NO: 58. In some embodiments, the 3’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to a nucleic acid sequence of SEQ ID NO: 58. In some embodiments, the 3’ UTR comprises a nucleic acid sequence of SEQ ID NO: 58 with one or more mutations therein. For example, in some embodiments, the 3’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 58. In some embodiments, the 3’ UTR comprises a nucleic acid sequence of SEQ ID NO: 58 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0094] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 5’ UTR and a 3’ UTR. In some embodiments, the 5’ UTR comprises or consists of a nucleic acid sequence selected from any one of SEQ ID NOs: 34-49; and the 3’ UTR comprises or consists of a nucleic acid sequence selected from any one of SEQ ID NOs: 50-61. In some embodiments, the 5’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or 100% sequence identity to a nucleic acid sequence selected from any one of SEQ ID NOs: 34-49; and the 3’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or 100% sequence identity to a nucleic acid sequence selected from any one of SEQ ID NOs: 50-61. In some embodiments, the 5’ UTR comprises a nucleic acid sequence selected from any one of SEQ IDAttorney Docket No. TORQ-012 / 02WO 339010-2071NOs: 34-49 with one or more mutations therein; and the 3 ’ UTR comprises a nucleic acid sequence selected from any one of SEQ ID NOs: 50-61 with one or more mutations therein. For example, in some embodiments, the 5’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in any one of SEQ ID NOs: 34-49; and the 3 ’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in any one of SEQ ID NOs: 50-61. In some embodiments, the 5’ UTR comprises a nucleic acid sequence selected from any one of SEQ ID NO: 34-49 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations; and the 3’ UTR comprises a nucleic acid sequence selected from any one of SEQ ID NO: 50-61 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0095] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 5’ UTR and a 3’ UTR. In some embodiments, the 5’ UTR comprises or consists of a nucleic acid sequence of SEQ ID NO: 44; and the 3’ UTR comprises or consists of a nucleic acid sequence of SEQ ID NO: 50. In some embodiments, the 5’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or 100% sequence identity to a nucleic acid sequence of SEQ ID NO: 44; and the 3’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or 100% sequence identity to a nucleic acid sequence of SEQ ID NO: 50. In some embodiments, the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 44 with one or more mutations therein; and the 3’ UTR comprises a nucleic acid of SEQ ID NO: 50 with one or more mutations therein. For example, in some embodiments, the 5’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 44; and the 3’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 50. In some embodiments, the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 44 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations; and the 3’ UTR comprises a nucleic acid sequence of SEQ ID NO: 50 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0096] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a 5’ UTR and a 3’ UTR. In some embodiments, the 5’ UTR comprises or consists of a nucleic acid sequence of SEQ ID NO: 37; and the 3’ UTR comprises or consists of a nucleic acid sequence ofAttorney Docket No. TORQ-012 / 02WO 339010-2071SEQ ID NO: 61. In some embodiments, the 5’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or 100% sequence identity to a nucleic acid sequence of SEQ ID NO: 37; and the 3’ UTR comprises a nucleic acid sequence with at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or 100% sequence identity to a nucleic acid sequence of SEQ ID NO: 61. In some embodiments, the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 37 with one or more mutations therein; and the 3’ UTR comprises a nucleic acid of SEQ ID NO: 61 with one or more mutations therein. For example, in some embodiments, the 5’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 37; and the 3’ UTR comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations in SEQ ID NO: 61, In some embodiments, the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 37 with 1 -5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations; and the 3’ UTR comprises a nucleic acid sequence of SEQ ID NO: 61 with 1-5, 5-10, 11-15, 15-20, 20-25, 25-30, or more than 30 mutations.

[0097] In some embodiments, the nucleic acid molecule encoding PC2 protein comprises a translation initiation sequence (e.g., a Kozak sequence). In some embodiments, the translation initiation sequence comprises or consists of a nucleic acid sequence of SEQ ID NO: 20. In some embodiments, the translation initiation sequence comprises a nucleic acid sequence with at least about 70%, at least about 75%, at least about 85%, or at least about 92% sequence identity to a nucleic acid sequence of SEQ ID NO: 20. In some embodiments, the translation initiation sequence comprises a nucleic acid sequence of SEQ ID NO: 20 with one or more mutations therein. For example, in some embodiments, the translation initiation sequence comprises 1, 2, 3, 4, 5, 6, or more mutations in SEQ ID NO: 20.

[0098] In some embodiments, the polynucleotide encoding the PC2 protein is operably linked to a sequence encoding a signal peptide for nuclear localization. In some embodiments, the signal peptide is an IgK signal peptide. In some embodiments, the signal peptide is a CDS signal peptide.

[0099] In some embodiments, the nucleic acid molecule of the present disclosure contains substitutions, additions, or deletions that alter the properties or activities of the encodedAttorney Docket No. TORQ-012 / 02WO 339010-2071polypeptide (i.e., PC2 protein). In some embodiments, the nucleic acid molecule of the present disclosure contains silent substitutions, additions, or deletions that does not alter the properties or activities of the encoded polypeptide (i.e., PC2 protein).

[0100] In some embodiments, a nucleic acid sequence variant is produced to modulate or alter expression (or expression levels) of the encoded polypeptide. In some embodiments, a nucleic acid sequence variant is produced to increase expression of the encoded polypeptide. In some embodiments, a nucleic acid sequence variant is produced to decrease expression of the encoded polypeptide. In some embodiments, a nucleic acid sequence variant has increased expression of the encoded polypeptide as compared to a parental nucleic acid sequence. In some embodiments, a nucleic acid sequence variant has decreased expression of the encoded polypeptide as compared to a parental nucleic acid sequence.

[0101] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide encoding the PC2 protein is codon-optimized. As used herein, the term “codon-optimized” refers to substituting codons in the polynucleotide encoding the PC2 protein in order to increase the expression, stability and / or activity of the PC2 protein. Factors that influence codon optimization include, but are not limited to one or more of: (i) variation of codon biases between two or more organisms or genes or synthetically constructed bias tables, (ii) variation in the degree of codon bias within an organism, gene, or set of genes, (iii) systematic variation of codons including context, (iv) variation of codons according to their decoding tRNAs, (v) variation of codons according to GC %, either overall or in one position of the triplet, (vi) variation in degree of similarity to a reference sequence for example a naturally occurring sequence, (vii) variation in the codon frequency cutoff, (viii) structural properties of mRNAs transcribed from the DNA sequence, (ix) prior knowledge about the function of the DNA sequences upon which design of the codon substitution set is to be based, (x) systematic variation of codon sets for each amino acid, (xi) isolated removal of spurious translation initiation sites, (xii) elimination of fortuitous polyadenylation sites otherwise leading to truncated RNA transcripts, (xiii) elimination of fortuitous predicted splice sites, and / or (xiv) disruption of CpG dinucleotides. In some embodiments, the polynucleotide encoding the PC2 protein is codon-optimized for expression in a human subject. In some embodiments, the polynucleotide encoding the PC2 protein is codon-optimized for expression in a mouse subject. In some embodiments, the polynucleotide encoding the PC2 protein is codon-optimized forAttorney Docket No. TORQ-012 / 02WO 339010-2071expression in a human subject and a mouse subject. In some embodiments, the polynucleotide encoding the PC2 protein is codon-optimized for expression in the kidney of a human subject. In some embodiments, the polynucleotide encoding the PC2 protein is codon-optimized for expression in the kidney of a mouse subject. In some embodiments, the polynucleotide encoding the PC2 protein is codon-optimized for expression in the kidney of a human subject and a mouse subject.

[0102] It will be appreciated by those of ordinary skill in the art that, as a result of the degeneracy of the genetic code, there are many nucleic acid sequences that encode a polypeptide (e.g., PC2 protein). Some of these nucleic acid sequences bear minimal homology to the nucleic acid sequences of the native gene. Nonetheless, nucleic acid sequences that vary due to differences in codon usage are specifically contemplated in particular embodiments, for example, nucleic acid sequences that are optimized for human and / or primate codon selection. Further, alleles of the genes comprising the nucleic acid sequences provided herein may also be used. Alleles are endogenous genes that are altered as a result of one or more mutations, such as deletions, additions and / or substitutions of nucleotides,

[0103] The nucleic acid molecules disclosed herein can be prepared, isolated, purified, manipulated, and / or expressed using any of a variety of well-established techniques known and available in the art.

[0104] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any one of SEQ ID NOs: 65-107 and 156-160. In some embodiments, the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 65-107 and 156-160 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 65-107 and 156-160. In some embodiments, the polynucleotide consists of a nucleic acid sequence of any one of SEQ ID NOs: 65-107 and 156-160.

[0105] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that isAttorney Docket No. TORQ-012 / 02WO 339010-2071at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 66. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 66 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 66. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 66.

[0106] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 158. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 158 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 158. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 158.

[0107] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 67. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 67 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 67. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 67.

[0108] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identicalAttorney Docket No. TORQ-012 / 02WO 339010-2071to SEQ ID NO: 159. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 159 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 159. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 159.

[0109] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any one of SEQ ID NOs: 108-155. In some embodiments, the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 108-155 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 108-155. In some embodiments, the polynucleotide consists of a nucleic acid sequence of any one of SEQ ID NOs: 108-155.

[0110] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 109. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 109 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 109. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 109.

[0111] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 153. In some embodiments, the polynucleotide comprises a nucleic acid sequenceAttorney Docket No. TORQ-012 / 02WO 339010-2071of SEQ ID NO: 153 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 153. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 153.

[0112] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 110, In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 110 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 110. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 110.

[0113] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a PC2 protein. In some embodiments, the polynucleotide comprises a nucleic acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 154. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 154 with one or more mutations, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations. In some embodiments, the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 154. In some embodiments, the polynucleotide consists of a nucleic acid sequence of SEQ ID NO: 154.

[0114] Exemplary recombinant nucleic acid molecules, and their corresponding elements, are provided in Table 1 below.Attorney Docket No. TORQ-012 / 02WO 339010-2071Table 1. Exemplary Cassettes of the Present DisclosureCassette SEQ ID NOs of ComponentsSEQPlasmid 5’ 5’ 3’ Term3’ID ProName Kozak CDSID ITR moter UTR UTR inator ITR NO PTR089 65 5ITR145-CBA-PKD2-3XHA-bGHpA-3ITR145 16 12 N / A 20 6 N / A 18 17 PTR087 66 5ITR145-CBA-PKD2coAcpg-3XHA-bGHpA-3ITR145 16 12 N / A 20 9 N / A 18 175ITR145-hPGK-PKD2coAcpg-3XHA-bGHpA-3ITR145 67 16 13 N / A 20 9 N / A 18 17 PTR1015ITR145 -hPGK-PKD2co Acpg-3XHA-2xmiRl 22 v 1 - PTR329 68 16 13 N / A 20 9 N / A 18 17 bGHpA-3ITR1455ITR145 -CBA-PKD2co Acpg-3XHA-2xmiRl 22v 1 -bGHpA- PTR330 69 16 12 N / A 20 9 N / A 18 173ITR1455ITR145-hEFla-PKD2coAcpg-3XHA-bGHpA-3ITR145 PTR331 70 16 21 N / A 20 9 N / A 18 17 5ITR145-mECAD-PKD2coAcpg-3XHA-bGHpA-3ITR145 PTR332 71 16 22 N / A 20 9 N / A 18 17 5ITR 145 -m mi -hWNKl -RP-PKD2co Acpg-3XHA-bGHpA- PTR333 72 16 23 N / A 20 9 N / A 18 173ITR1455ITR145-mKSPC-PKD2coAcpg-3XHA-rBGHpA-3ITR145 PTR334 73 16 24 N / A 20 9 N / A 33 17 5ITR145-RP-hPGK-PKD2coAcpg-3XHA-rBGHpA- 74 16 25 N / A 20 9 N / A 33 17 PTR3363ITR1455ITR145-mPAX8-PKD2coAcpg-3XHA-bGHpA-3ITR145 PTR338 75 16 26 N / A 20 9 N / A 18 17 5ITR145-hPKD2-PKD2coAcpg-3XHA-bGHpA-3ITR145 PTR339 76 16 27 N / A 20 9 N / A 18 17 5ITR145-hPKD2-G4ml-PKD2coAcpg-3XHA-bGHpA- PTR340 77 16 28 N / A 20 9 N / A 18 173 ITR 1455ITR145-pPKD2-PKD2coAcpg-3XHA-bGHpA-3ITR145 PTR341 78 16 29 N / A 20 9 N / A 18 17 5ITR145-pPKD2-G4m 1 -PKD2coAcpg-3XHA-bGHpA- PTR342 79 16 30 N / A 20 9 N / A 18 173ITRI455ITR145-mPKD2-483-PKD2coAcpg-3XHA-bGHpA- PTR343 80 16 31 N / A 20 9 N / A 18 173 ITR 1455 ITR145 -mPKD2-389-PKD2co Acpg-3XHA-bGHpA- 20PTR344 81 16 32 N / A 9 N / A 18 173 ITR 1455ITR145-hPGK-5utr9765-PKD2coAcpg-3XHA-bGHpA- PTR284 82 16 13 34 20 9 N / A 18 1731TR145Attorney Docket No. TORQ-012 / 02WO 339010-2071Cassette SEQ ID NOs of ComponentsSEQPlasmid 5’ Pro5’ 3’ Term3’ID Name Kozak CDSID ITR moter UTR UTR inator ITR NO5ITR145-hPGK-5utrll674-PKD2coAcpg-3XHA-bGHpA- PTR285 83 16 13 35 20 9 N / A 18 173ITR1455ITR145-hPGK-5utrl0023-PKD2coAcpg-3XHA-bGHpA- PTR286 84 16 13 36 20 9 N / A 18 173ITR1455ITR145-hPGK-5utrhHBB-PKD2coAcpg-3XHA- PTR290 85 16 13 37 20 9 50 18 173utrhHBB-bGHpA-3ITR1455ITR145-hPGK-5utrPoV-pA-scrUTR-PKD2coAcpg- PTR291 86 16 13 38 20 9 50 18 173XHA-3utrhHBB-bGHpA-3ITR1455ITR145-hPGK-5utrTOP-PKD2coAcpg-3XHA-3utrhHBB- PTR292 87 16 13 39 20 9 50 18 17 bGHpA-3ITR1455ITR145-hPGK-5utrTMV-PKD2coAcpg-3XHA-3utrhHBB- PTR293 88 16 13 40 20 9 50 18 17 bGHpA-3ITR1455ITR145-hPGK-5utrTEV-PKD2coAcpg-3XHA-3utrhHBB- PTR294 89 16 13 41 20 9 50 18 17 bGHpA-3ITR1455ITR145-hPGK-5utrscrUTR-PKD2coAcpg-3XHA- PTR295 90 16 13 42 20 9 50 18 173utrhHBB-bGHpA-3ITR1455ITR145-hPGK-5utrRpS25-PKD2coAcpg-3XHA- PTR296 91 16 13 43 20 9 50 18 173utrhHBB-bGHpA-3ITR1455ITR 145 -hPGK-5utrRpL38 -PKD2co Acpg-3XH A- PTR297 92 16 13 44 20 9 50 18 173utrhHBB-bGHpA-3ITR1455ITR145-hPGK-5utrmActb-PKD2coAcpg-3XHA- PTR298 93 16 13 45 20 9 50 18 173utrhHBB-bGHpA-3ITR1455ITR145-hPGK-5utrC3-PKD2coAcpg-3XHA-3utrbHBB- PTR299 94 16 13 46 20 9 50 18 17 bGHpA-3ITRI455ITR145-hPGK-5utrhACTB-PKD2coAcpg-3XHA- PTR300 95 16 13 47 20 9 50 18 173utrhHBB-bGHpA-3ITRl 455ITR145 -hPGK-5 utrhHBB-PKD2co Acpg-3XHA- PTR301 96 16 13 37 20 9 53 18 173utrCYBA-l,5x-bGHpA-3ITR1455ITR145 -hPGK-5 utrhHBB-PKD2co Acpg-3XHA- PTR302 97 16 13 37 20 9 54 18 173utrCYBA-bGHp A-3ITR 145Attorney Docket No. TORQ-012 / 02WO 339010-2071Cassette SEQ ID NOs of ComponentsSEQPlasmid 5’ Pro5’ 3’ Term3’ID Name Kozak CDSID ITR moter UTR UTR inator ITR NO5ITR145-hPGK-5utrhIIBB-PKD2coAcpg-3XHA- PTR303 98 16 13 37 20 9 55 18 173utrhHBAl-bGHpA-3ITR1455ITR145-hPGK-5utrhIIBB-PKD2coAcpg-3XHA- PTR304 99 16 13 37 20 9 56 18 173utrhHBBx2-bGHpA-3ITR1455ITR145-hPGK-PKD2coAcpg-3XHA-3utrHuR-BS- PTR305 100 16 13 N / A 20 9 57 18 17 bGHpA-3ITR145PTR306 BB003 5ITR145-hPGK-PKD2coAcpg-3XHA- PTR306 101 16 13 N / A 20 9 58 18 173utrHuR-BSx3-bGHpA-3ITRl 455ITR145-bPGK-PKD2coAcpg-3XHA-3utrCD47partial3'-A- PTR307 102 16 13 N / A 20 9 59 18 17 bGHpA-3ITR1455ITR145-bPGK-PKD2coAcpg-3XHA-3utrCD47partial3'-B- PTR308 103 16 13 N / A 20 9 60 18 17 bGHpA-3ITR1455ITR145-hPGK-5utrhHBB-PKD2coAcpg-3XHA- PTR309 104 16 13 37 20 9 61 18 173utrDEN2-bGHpA-3ITR1455ITR145-hPGK-5utrmRpll8a-P4-mActb-PKD2coAcpg- PTR310 105 16 13 48 20 9 50 18 173XHA-3utrhHBB-bGHpA-3ITR1455ITR145-hPGK-5utrP4-mActb-PKD2coAcpg-3XHA- PTR311 106 16 13 49 20 9 55 18 173utrhHBAl-bGHpA-3ITR1455ITR145-hPGK-PKD2coAcpg-3XHA-5xmiR122vl- PTR327 107 16 13 N / A 20 9 52 18 17 bGHpA-3ITR145PTR095 156 5ITR145-hPGK-PKD2Acpg-3XHA-bGHpA-3ITR145 16 13 N / A 20 8 N / A 18 17 PTR099 157 5ITR145-CBA-PKD2Acpg-3XHA-bGHpA-3ITR145 16 12 N / A 20 8 N / A 18 17 PTR108 158 5ITR145-CBA-PKD2-bGHpA-3ITR145 16 12 N / A 20 2 N / A 18 17 PTR109 159 5ITR145-hPGK-PKD2coAcpg-bGHpA-3ITR145 16 13 N / A 20 5 N / A 18 175ITR145-CBA-PKD2coAcpg-bGHpA-3ITR145PTR134 160 16 12 N / A 20 5 N / A 18 17 N / A = Not ApplicableAttorney Docket No. TORQ-012 / 02WO 339010-2071Non- Viral and Viral Vectors

[0115] In some embodiments, a vector comprises the nucleic acid molecule encoding the PC2 protein described herein. In some embodiments, the vector is double-stranded. In some embodiments, the vector is single-stranded. In some embodiments, the vector is self- complementary. In some embodiments, the vector is a non-viral vector. In some embodiments, the vector is a viral vector.

[0116] In some embodiments, the vector is a viral vector. A number of viral based systems have been developed for gene transfer into mammalian cells. Examples of viral vectors include, but are not limited to, adenoviral vectors, adeno-associated virus vectors, lentiviral vectors, retroviral vectors, herpes simplex viral vectors, y-retroviral vectors, arenavirus vectors, alphavirus vectors, baculovirus vectors, vaccina virus vectors, parvovirus vectors, and derivatives thereof. Viral vector technology is well known in the art and is described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and in other virology and molecular biology manuals. In some embodiments, the viral vector is a recombinant adeno-associated virus (rAAV) vector.

[0117] In some embodiments, the AAV vector is a chimeric AAV vector, a single-stranded AAV vector, or a self-complementary AAV vector. In some embodiments, the viral vector is a specific AAV serotype. In some embodiments, the viral vector is a human AAV serotype.

[0118] In some embodiments, the AAV vector is a self-complementary AAV (scAAV) vector. Self-complementary AAV vectors contain complementary sequences that spontaneously anneal to form a double-stranded DNA genome when entering a target cell.

[0119] In some embodiments, the viral vector is a pseudotyped AAV vector. A pseudotyped AAV vector, as used herein, refers to a genetically engineered vector derived from AAV that has been modified by incorporating the capsid from a different AAV serotype or virus. In some embodiments, a pseudotyped AAV vector comprises an AAV backbone derived from one serotype and an AAV capsid derived from another serotype. Specific examples of such pseudotyped AAV vectors, include, but are not limited to, vectors comprising an AAV2-derived genome with an AAV1-derived capsid; an AAV2-derived genome with an AAV4-derived capsid; an AAV2-derived genome with an AAV5-derived capsid; an AAV2-derived genome with an AAV8-derived capsid; or an AAV2-derived genome with an AAV9-derived capsid. In some embodiments, the capsid is derived from another type of virus, such as a parvovirus.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0120] In some embodiments, the viral vector is a chimeric viral vector (e.g., a chimeric AAV vector). As used herein, the term “chimeric viral vector” refers to a genetically engineered vector derived from a virus that has been modified by incorporating genetic material from other viruses or organisms. For example, in some embodiments, an AAV vector incorporates elements from different AAV serotypes or other viruses to overcome the limitations of the original AAV vector, such as pre-existing immunity in the host population. In some embodiments, the chimeric viral vector comprises viral portions from two or more viruses. In some embodiments, the chimeric viral vector is genetically engineered to increase transduction efficiency, selectivity, or a combination thereof. In some embodiments, the chimeric viral vector comprises capsid proteins from two or more serotypes. Chimeric viral vectors are further described in International Patent Appl. No. WO 2000 / 028004.

[0121] The rAAV vector can be derived from any AAV serotypes or variants thereof. In some embodiments, the rAAV vector is derived from AAV1 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV2 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV2i8 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV3 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV3-B or a variant thereof. In some embodiments, the rAAV vector is derived from AAV4 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV 5 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV6 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV7 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV8 or a variant thereof. In some embodiments, the rAAV vector is derived from AAVrh8 or a variant thereof. In some embodiments, the rAAV vector is derived from AAVrh8R or a variant thereof. In some embodiments, the rAAV vector is derived from AAV9 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV 10 or a variant thereof. In some embodiments, the rAAV vector is derived from AAVrhlO or a variant thereof. In some embodiments, the rAAV vector is derived from AAV 11 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV12 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV13 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV-DJ or a variant thereof. In some embodiments, the rAAV vector is derived from AAV LK03 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV KP1 or a variant thereof. In some embodiments, the rAAV vector is derivedAttorney Docket No. TORQ-012 / 02WO 339010-2071from AAV.cc47 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV.cc84 or a variant thereof. In some embodiments, the rAAV vector is derived from AAVrh74 or a variant thereof. In some embodiments, the rAAV vector is derived from AAV44-9 or a variant thereof. In some embodiments, the rAAV vector is derived from avian AAV, bovine AAV, canine AAV, equine AAV, ovine AAV, and any combination, derivative, or variant thereof. See also, Fields et al., Virology, Vol. 2, 4th edition; and Gao et al., (2004) J Virology 78:6381-6388; and Moris et al., Virology 33:375-383, each of which are herein incorporated by reference in their entirety.

[0122] In some embodiments, the rA AV vectors provided herein comprise a modification, such as an insertion, deletion, chemical alteration, or synthetic modification, relative to a wild-type AAV vector.

[0123] In some embodiments, the rA AV vectors described herein comprise one or more AAV backbone elements. As used herein, an “AAV backbone element” refers to AAV genomic elements required for the bioactivity of AAV vectors. A AV backbone elements include, but are not limited to, packaging elements for the A AV vector to be assembled into an AAV particle, viral replication elements, reporter elements, and elements for expression of the circular RNA-encoding sequences described herein.

[0124] In some embodiments, the rAAV vectors described herein comprise one or more inverted terminal repeats (ITRs). An ITR is a specific DNA sequence found in adeno-associated viruses that is crucial for viral replication, packaging, and stability of the AAV genome. In some embodiments, the rAAV vector comprises a 5’ inverted terminal repeat (ITR) and a 3’ ITR. In some embodiments, the 5 ’ ITR flanks the 5 ’ end of the nucleic acid molecule and the 3 ’ I TR flanks the 3 ’ end of the nucleic acid molecule.

[0125] In some embodiments, the 5’ and / or 3’ ITRs used in the rAAV vectors described herein have a wild- type nucleotide sequence. In some embodiments, the 5’ and / or 3’ ITRs used in the vectors described herein are not wild-type, but instead, comprise, e.g., an insertion, deletion or substitution of one or more nucleotides. AAV ITRs provided herein may be derived from any AAV serotype, including, but not limited to, AAV1, AAV2, AAV2i8, AAV3, AAV3-B, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh8R, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAV-DJ, AAV LK03, AAV KP1, AAV.cc47, AAV.84, AAVrh74, AAV44-9, or a variant thereof.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0126] In some embodiments, the 5’ and 3’ ITRs which flank the nucleic acid molecule provided herein are identical and derived from the same AAV serotype. In some embodiments, the 5’ and 3’ ITRs which flank the nucleic acid molecule provided herein are different and / or derived from different AAV serotypes.

[0127] In some embodiments, the AAV vectors described herein comprise a Flip and / or Flop ITR. The term “Flip ITR” or “Flop ITR,” as used herein, refers to the orientation of the ITR determined by the arrangement of its palindromic sequences. A single ITR comprises palindromic arms (A-A’, B-B’, and C-C’) that give the ITR its characteristic T-shape. The arrangement of the B-B’ and C-C’ palindromic sequences determines the orientation of the ITR, which can be either in the “Flip’ or “Flop” orientation. In some embodiments, the AAV vector comprises a 5’ Flip ITR and a 3’ Flop ITR. In some embodiments, the AAV vector comprises a 5’ Flop ITR and a 3’ Flip ITR, In some embodiments, the AAV vector comprises a 5’ Flip ITR and a 3’ Flip ITR. In some embodiments, the AAV vector comprises a 5’ Flop ITR and a 3’ Flop ITR. In some embodiments, the 5’ and 3’ Flip and / or Flop ITRs are derived from the same AAV serotype. In some embodiments, the 5’ and 3’ Flip and / or Flop ITRs are derived from a different AAV serotype.

[0128] In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 80% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 85% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 90% identical to SEQ ID NO: 16, In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 91% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 92% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 93% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 94% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 95% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 96% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is at least about 98% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence that is atAttorney Docket No. TORQ-012 / 02WO 339010-2071least about 99% identical to SEQ ID NO: 16. In some embodiments, the 5’ ITR comprises a nucleic acid sequence of SEQ ID NO: 16.

[0129] In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 80% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 85% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 90% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 91% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 92% identical to SEQ ID NO: 17. In some embodiments, the 3 ’ ITR comprises a nucleic acid sequence that is at least about 93% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 94% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 95% identical to SEQ ID NO: 17, In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 96% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 17, In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 98% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises a nucleic acid sequence of SEQ ID NO: 17.

[0130] In some embodiments, the rAAV vector comprising the nucleic acid molecule flanked by AAV ITRs can be constructed by directly inserting the nucleic acid molecule into an AAV genome, e.g., into an excised AAV open reading frames, and certain portions of the AAV genome can optionally be deleted, as described in, e.g., WO 1993 / 003769; Kotin (1994) Human Gene Therapy 5: 793-801; Shelling and Smith (1994) Gene Therapy 1: 165-169; and Zhou et al. (1994) J. Exp. Med. 179: 1867-1875.

[0131] In some embodiments, AAV ITRs are excised from an AAV genome or from an AAV vector containing such ITRs, and then are fused to 5’ and 3’ of the nucleic acid molecule that is present in another vector using standard ligation techniques.Recombinant Viral Particles

[0132] In some embodiments, the viral vector genomes described herein are packaged into viral particles, which are used to deliver the nucleic acid molecule encoding a PC2 protein to target cells.Attorney Docket No. TORQ-012 / 02WO 339010-2071Viral particles of the present invention can be produced using any method known in the art, e.g., expression from a baculovirus (Brown et al. (1994) Virology 198:477-488). In some embodiments, the viral vector genome is an AAV vector genome packaged into an AAV particle.

[0133] In some embodiments, methods of producing AAV particles herein comprise packaging a nucleic acid molecule encoding a PC2 protein into an AAV vector. In some embodiments, methods of producing the AAV particle comprises: (a) contacting a cell with at least one nucleic acid molecule encoding: (i) a PC2 protein; (ii) a replication gene; and (hi) a capsid gene that encodes an AAV capsid protein; (b) expressing the AAV capsid protein in the cell; (c) assembling an AAV particle; and (d) packaging the nucleic acid encoding the PC2 protein into the AAV particle.

[0134] In some embodiments, the present disclosure provides a method of expressing a PC2 protein in a cell by introducing the AAV vector or AAV capsid or particle comprising a nucleic acid molecule encoding the PC2 protein into the cell under conditions wherein the nucleic acid molecule is transcribed and translated to produce the polycystin-2 protein. In some embodiments, the cell is a mammalian cell. In some embodiments, the cell is a human cell. In some embodiments, the cell is a myoblast. In some embodiments, the cell is a kidney cell. In some embodiments, the cell is a kidney cell derived from or localized in the thin ascending limb, connecting tubule, collecting duct, thin descending limb, distal tubule, proximal tubule, Loop of Henle, glomerulus, or thick ascending limb. In some embodiments, the kidney cell is an endothelial cell, a mesangial cell, a glomerular endothelial cell, a parietal cell, a proximal tubule cell, a descending limb cell, a thin and / or thick ascending limb cell, a convoluted tubule cell, a connecting tubule cell, a fibroblast, an immune cell (e.g., a natural killer cell, a T cell, a B cell, a macrophage, or a monocyte), an intercalated cell, a parietal epithelial cell, a podocyte, a principal cell, a Schwann cell, a vascular smooth muscle cell, or a pericyte.

[0135] In general, an AAV capsid comprises three proteins, VP1, VP2 and VP3, each of which is encoded by splice variants of the AAV cap gene. VP2 and VP3 are truncated version of VP1, and thus have sequences that are also comprised by VP1. Generally, the amino acid sequence of VP1 defines the serotype of the capsid. Thus, for example, if the VP1 capsid protein encodes for an AAV2 VP1 protein, AAV will be of the AAV2 serotype, whereas if the VP1 capsid protein encodes an AAV8 VP1 protein, the AAV will be of the AAV8 serotype.

[0136] In some embodiments, the AAV capsid protein is a VP1 capsid protein. In some embodiments, the AAV capsid protein is a VP2 capsid protein. In some embodiments, the AAVAttorney Docket No. TORQ-012 / 02WO 339010-2071capsid protein is a VP3 capsid protein. In some embodiments, the rAAV particle comprises a VP1 capsid protein, a VP2 capsid protein, and / or a VP3 capsid protein. In some embodiments, the rAAV particle comprises a VP1 capsid protein, a VP2 capsid protein, and / or a VP3 capsid protein, wherein the capsid proteins of the rAAV particle are of the same serotype. In some embodiments, the rAAV particle comprises a VP1 capsid protein, a VP2 capsid protein, and a VP3 capsid protein, wherein the capsid proteins of the AAV particle are of the same serotype.

[0137] In some embodiments, an AAV capsid protein (e.g., VP1, VP2 and / or VP3) in the present rAAV particle is not a naturally occurring capsid protein. In some embodiments, an AAV capsid protein (e.g., VP1, VP2 and / or VP3) is derived from a naturally occurring capsid protein.

[0138] In some embodiments, the capsid protein is a capsid protein of A AVI or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV2 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV2i8 or a variant thereof, In some embodiments, the capsid protein is a capsid protein of AAV3 or a variant thereof, In some embodiments, the capsid protein is a capsid protein of A AV3-B or a variant thereof, In some embodiments, the capsid protein is a capsid protein of AAV4 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV5 or a variant thereof. In some embodiments. the capsid protein is a capsid protein of AAV6 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV7 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV8 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAVrh8 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAVrh8R or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV9 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV10 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAVrhlO or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV11 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV 12 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV13 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV-DJ or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV LK03 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV KP1 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV.cc47 or a variant thereof. In someAttorney Docket No. TORQ-012 / 02WO 339010-2071embodiments, the capsid protein is a capsid protein of AAV.cc84 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAVrh74 or a variant thereof. In some embodiments, the capsid protein is a capsid protein of AAV44-9 or a variant thereof. In some embodiments, the capsid is an AAV9 capsid variant, such as AAV.klS or AAV.k20, as described in PCT Publication No. WO / 2024 / 206226, which is incorporated herein by reference in its entirety.

[0139] The capsid protein can be derived from the same serotype as the ITRs or a variant thereof. The capsid protein can also be of a different serotype than the ITR.

[0140] In some embodiments, the AAV vectors described herein comprise a modified capsid to alter the tropism of the vector. In some embodiments, the modified capsid comprises proteins or peptides of non-viral origin to alter the tropism of the vector. For example, in some embodiments, the capsid comprises a ligand of a particular receptor, or a receptor of a particular ligand, to target the vector to the cell type(s) expressing said receptor or ligand, respectively. In some embodiments, the modified capsid is altered to target the vector to a particular tissue.

[0141] In some embodiments, the AAV capsid is a variant of a parental wild-type AAV capsid, wherein the AAV capsid improves transfer and / or expression of the recombinant nucleic acid molecule described herein in one or more region(s) or part(s) of the kidney compared to the parental wild-type AAV capsid.

[0142] Exemplary AAV capsids with improved transfer and / or expression in kidney cells and kidney-related cells, as well as methods of using AA V particles comprising such AAV capsids to efficiently deliver a cargo to one or more region(s) or part(s) of kidney is disclosed in PCT Publication No. WO / 2024 / 206226.

[0143] In some embodiments, the AAV capsid comprises one or more mutations, e.g., one or more amino acid substitutions, amino acid deletions, or heterologous peptide insertions, compared to a parental wild-type AAV capsid, such as a naturally-occurring capsid protein from which it was derived. In some embodiments, the ammo acid sequence of the AAV capsid (e.g., VP1, VP2, and / or VP3) is identical to the ammo acid sequence of the parental wild-type AAV capsid except for 1, 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 or 30 amino acid residues, e.g., except for 1, 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 or 30 amino acid residue substitutions. In some embodiments, the AAV capsid is an AAV.kl3 capsid. In some embodiments, the AAV capsid is an AAV.k20 capsid. Exemplary' AAV capsids with improved transfer and / or expression in kidneyAttorney Docket No. TORQ-012 / 02WO 339010-2071cells and kidney-related cells, as well as methods of using AAV particles comprising such AAV capsids to efficiently deliver a cargo to one or more regions or parts of kidney is disclosed in PCT Publication No. WO / 2024 / 206226.

[0144] In some embodiments, the rAAV comprises a vector comprising a transgene and an AAV capsid. In some embodiments, the AAV capsid is an AAV.klS capsid. In some embodiments, the AAV capsid comprises an amino acid sequence of SEQ ID NO: 63 with 1, 2, 3, 4, or more amino acid substitutions at positions 452-458, wherein positions 452-458 of the AAV capsid are numbered with reference to SEQ ID NO: 62. In some embodiments, the AAV capsid comprises the amino acid sequence of SEQ ID NO: 63 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62. In some embodiments, the A AV capsid protein consists of the amino acid sequence of SEQ ID NO: 63 at positions 452-458, wherein positions 452-458 of the A AV capsid protein are numbered with reference to SEQ ID NO: 62,

[0145] In some embodiments, the rAAV comprises a vector comprising a transgene and an AAV capsid. In some embodiments, the AAV capsid is an AAV.k20 capsid. In some embodiments, the AAV capsid comprises an amino acid sequence of SEQ ID NO: 64 with 1, 2, 3, 4, or more amino acid substitutions at positions 452-458, wherein positions 452-458 of the AAV capsid are numbered with reference to SEQ ID NO: 62. In some embodiments, the AAV capsid comprises the amino acid sequence of SEQ ID NO: 64 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62. In some embodiments, the AAV capsid protein consists of the amino acid sequence of SEQ ID NO: 64 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

[0146] Exemplary ammo acid sequences for AAV9 capsid variants are provided in Table 2 below. For wild-type AAV9, bolded residues indicate position 452-458 m the AAV9 VPl subunit.Attorney Docket No. TORQ-012 / 02WO 339010-2071Table 2. AAV CapsidsDescription Amino Acid Sequence SEQ ID NO Wild-type MAADGYLPDWLEDNLSEGIREWWALKPGAI’QPKANQ 62AAV9 QHQDNARGLVLPGYKYLGPGNGLDKGEPVNAADAAA LEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDT SFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRP VEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVP DPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADG VGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLY KQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSP RDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVK TIANNLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPAD VFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTG NNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYI.. YY LSKTINGSGQNQQTLKFSVAGPSNMAVQGRNYIPGPS YRQQR VS TT VTQNNNSEF AWPGAS S W ALNGRNSLMN PGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVDAD KVNnTNEEEIKTTNPVAIES YGQ VATNHQS AQ AQ AQT GWVQNQGIIJ’GMVWQDRDVYLQGPIWAKIPHTDGNF HPSPLMGGFGMKHPPPQIIJKNTPVPADPPTAFNKDKL NSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNY YKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL AAV.kl3 MVEGRMG 63AAV.k20 VSGRREG 64

[0147] In some embodiments, the AAV capsid transduces one or more region(s) or part(s) of a kidney. In some embodiments, the AAV capsid has tropism for kidney cells. In some embodiments, the AAV capsid has tropism for one or more region(s) or part(s) of a kidney. In some embodiments, the region or part of the kidney is adrenal glands. In some embodiments, the region or part of the kidney is the cortex. In some embodiments, the region or part of the kidney is the medulla. In some embodiments, the region or part of kidney is the renal column. In some embodiments, the region or part of the kidney is the renal pyramid. In some embodiments, the region or part of the kidney is the renal pelvis. In some embodiments, the region or part of the kidney is the major calyx. In some embodiments, the region or part of kidney is the minor calyx. In some embodiments, the region or part of the kidney is the papillae. In some embodiments, the region or part of the kidney is the ureter. In some embodiments, the region or part of kidney is the proximal tubule (e.g, proximal convoluted tubule) in the cortex. In some embodiments, the region or part of kidney is the distal tubule (e.g., a distal convoluted tubule). In some embodiments, the region or part of kidney is the collecting tubule. In some embodiments, the region or part of kidney is the Loop ofAttorney Docket No. TORQ-012 / 02WO 339010-2071Henle. In some embodiments, the region or part of the kidney is the thick ascending limb. In some embodiments, the region or part of kidney is the glomerulus. In some embodiments, the AAV capsid is an AAV.kl3 capsid. In some embodiments, the AAV capsid is an AAV.k20 capsid.

[0148] In some embodiments, the AAV capsid transduces kidney cells. In some embodiments, the kidney cell is a mesangial cell. In some embodiments, the kidney cell is a podocyte. In some embodiments, the kidney cell is a glomerular endothelial cell. In some embodiments, the kidney cell is a parietal cell. In some embodiments, the kidney cell is a proximal tubule cell. In some embodiments, the kidney cell is a descending limb cell. In some embodiments, the kidney cell is a thin ascending limb cell and / or a thick ascending limb cell. In some embodiments, the kidney cell is a distal convoluted tubule cell. In some embodiments, the kidney cell is a connecting tubule cell. In some embodiments, the kidney cell is a collecting duct cell. In some embodiments, the kidney cell is a principal cell. In some embodiments, the kidney cell is a intercalated cell. In some embodiments, the kidney cell is a vascular smooth muscle cell. In some embodiments, the kidney cell is an endothelial cell. In some embodiments, the kidney cell is a fibroblast. In some embodiments, the kidney cell is an immune cell. In some embodiments, the immune cell is a natural killer cell, a T cell, a B cell, a macrophage, and / or a monocyte. In some embodiments, the AAV capsid is an AAV.kl3 capsid. In some embodiments, the AAV capsid is an AAV.k20 capsid.

[0149] In some embodiments, the AAV capsid variant transduces one or more kidney cells, kidney-derived cell types, and / or kidney-related cell types more efficiently than that of the wild¬ type AAV capsid. In some embodiments, the AAV capsid variant increases or improves transduction efficiency in the kidney by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, or more compared to the wild-type AAV capsid. In some embodiments, the kidney cell is a proximal tubule cell, a distal tubule cell, and / or a collecting duct cell.

[0150] In some embodiments, the AAV.kl3 capsid transduces one or more kidney cells, kidney-derived cell types, and / or kidney-related cell types more efficiently than that of the wild-type AAV9 capsid. In some embodiments, the AAV.kl3 capsid increases or improves transduction efficiency m the kidney by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least aboutAttorney Docket No. TORQ-012 / 02WO 339010-207145%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, or more, compared to the wild-type AAV9 capsid. In some embodiments, the kidney cell is a proximal tubule cell, a distal tubule cell, and / or a collecting duct cell.

[0151] In some embodiments, the AAV.k20 capsid transduces one or more kidney cells, kidney- derived cell types, and / or kidney-related cell types more efficiently than that of the wild-type AAV9 capsid. In some embodiments, the AAV.k20 capsid increases or improves transduction efficiency in the kidney by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, or more, compared to the wild-type AAV9 capsid. In some embodiments, the kidney cell is a proximal tubule cell, a distal tubule cell, and / or a collecting duct cell.

[0152] The rAAV particles described herein may be produced using any suitable method known in the art. For example, a host cell (e.g., a mammalian cell) may be engineered to stably express the necessary components for AAV particle production. This can be achieved by integrating a plasmid (or multiple plasmids) comprising AAV rep and cap genes, and a selectable marker, such as an antibiotic (e.g., neomycin or ampicillin) resistance gene into the genome of the cell. The cell can be, e.g., an insect or mammalian cell which can then be co-infected with a helper virus (e.g., adenovirus or baculovirus providing the helper functions) and the rAAV vector comprising the 5’ and 3 ’ AAV HR. The use of a selectable marker allows for large-scale production of the rAAV. As another non-limiting example, adenovirus or baculovirus rather than plasmids can be used to introduce rep and cap genes into packaging cells. As yet another non-limiting example, both the viral vector containing the 5’ and 3’ AAV ITRs and the rep and cap genes can be stably integrated into the DNA of producer cells, and the helper functions can be provided by a wild-type adenovirus to produce the rAAV.

[0153] In some embodiments, host cells containing the rAAV vectors described above is rendered capable of providing AAV helper functions to replicate and encapsulate the polynucleotide encoding the protein of interest provided herein flanked by the AAV ITRs to produce rAAV particles. AAV helper functions are generally AAV-derived coding sequences which can be expressed to provide AAV gene products that, in turn, function in trans for productive AAVAttorney Docket No. TORQ-012 / 02WO 339010-2071replication. AAV helper functions are used herein to complement necessary AAV functions that are missing from the rAAV vectors. In some embodiments, AAV helper functions include one, or both of the major AAV ORFs, namely the rep and cap coding regions, or functional homologues thereof.

[0154] AAV helper functions can be introduced into the host cell by transfecting the host cell with an AAV helper construct either prior to, or concurrently with, the transfection of the rAAV vector. For example, AAV helper constructs can be used to provide at least transient expression of AAV rep and / or cap genes to complement missing AAV functions that are necessary' for productive AAV infection. Typically, AAV helper constructs lack AAV ITRs and can neither replicate nor package themselves. The AAV helper constructs can be in the form of, e.g., a plasmid, phage, transposon, cosmid, virus, or virion.

[0155] A helper virus for AAV refers to a virus that allows A AV to be replicated and packaged by a host cell. A helper virus provides helper functions that allow for the replication of A AV. A number of such helper viruses have been identified, including adenoviruses, herpesviruses and poxviruses such as vaccinia. The adenoviruses encompass a number of different subgroups, although Adenovirus type 5 of subgroup C (Ad5) is most commonly used. Numerous adenoviruses of human, non-human mammalian and avian origin are known and are available from depositories such as the ATCC. Viruses of the herpes family, which are also available from depositories such as ATCC, include, for example, herpes simplex viruses (HSV), Epstein-Barr viruses (EBV), cytomegaloviruses (CMV) and pseudorabies viruses (PRV). Examples of adenovirus helper functions for the replication of AAV include El A functions, E1B functions, E2A functions, V A functions and E4orf6 functions.

[0156] A preparation of AAV is said to be substantially free of helper virus if the ratio of infectious AAV particles to infectious helper virus particles is at least about 102: 1; at least about 104: 1, at least about 106: 1; or at least about 108: 1. Preparations can also be free of equivalent amounts of helper virus proteins (i.e., proteins as would be present as a result of such a level of helper virus if the helper virus particle impurities noted above were present in disrupted form). Viral and / or cellular protein contamination can generally be observed as the presence of Coomassie staining bands on SDS gels (e.g., the appearance of bands other than those corresponding to the AAV capsid proteins VP1, VP2 and VP3).Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0157] In some embodiments, the AAV replication and capsid genes are provided by any method known in the art. In some embodiments, the AAV replication and capsid genes are on a single vector. In some embodiments, the AAV replication and capsid genes are on separate vectors. In some embodiments, the vector(s) comprising the AAV replication and / or capsid genes are viral (e.g., AAV, herpesvirus, EBV) or non-viral vectors. In some embodiments, the vector(s) comprising the AAV replication and / or capsid genes are AAV vectors. In some embodiments, the vector(s) comprising the AAV replication and / or capsid genes are chimeric AAV vectors (e.g., inserted into the El A or E3 regions of a deleted adenovirus vector).

[0158] In some embodiments, the host cell is also capable of providing or is provided with non AAV-derived functions or “accessory functions” to produce rAAV particles. Accessory functions are non AAV-derived viral and / or cellular functions upon w'hich AAV is dependent for its replication, such as non AAV proteins and RNAs that are required in AA V replication, including those involved in activation of AAV gene transcription, stage specific AAV mRNA splicing, AAV DNA replication, synthesis of Cap expression products and AAV capsid assembly. In some embodiments, viral-based accessory functions can be derived from a known helper virus.

[0159] A variety of host cells can be used to produce rAAV particles described herein. Suitable host cells for producing AAV particles from the nucleic acid molecules and AAV vectors provided herein include microorganisms, yeast cells, insect cells, and mammalian cells. Typically, such cells can be, or have been, used as recipients of a heterologous nucleic acid molecule and can grow in, e.g., suspension culture and a bioreactor.

[0160] In some embodiments, the cell is a mammalian host cell, for example, a HEK293, HEK293-T, A549, WEHI, 10T1 / 2, BHK, MDCK, COS1, COS7, BSC 1, BSC 40, BMT 10, VERO, W138, HeLa, 293, Jurkat, 2V6.11, Saos, C2C12, L, HIT 080, HepG2, primary fibroblast, hepatocyte, and myoblast cells.

[0161] In some embodiments, the cell is an insect cell, for example an Sf9, SF21, SF900+, or a drosophila cell lines, mosquito cell lines, e.g., Aedes albopictus derived cell lines, domestic silkworm cell lines, e.g., Bombyxmori cell lines, Tnchoplusia ni cell lines such as High Five cells or Lepidoptera cell lines such as Ascalapha odorata cell lines. In some embodiments, insect cells are cells from the insect species which are susceptible to baculovirus infection, including High Five, Sf9, Se301, SeIZD2109, SeUCRl, Sf900+, Sf21, BTI-TN-5B1-4, MG-1, Tn368, HzAml, BM-N, Ha2302, Hz2E5 and Ao38. For example, large scale production of recombinant AAV inAttorney Docket No. TORQ-012 / 02WO 339010-2071cells, including Sf9 insect cells, has been described by Kotin RM. Hum Mol Genet. 20 (Rl): R2-R6 (2011) doi: 10.1093 / hmg / ddrl41. Methodology for molecular engineering and expression of polypeptides in insect cells is described, for example, in Summers and Smith. A Manual of Methods for Baculovirus Vectors and Insect Culture Procedures, Texas Agricultural Experimental Station Bull. No. 7555, College Station, Tex. (1986); King, L. A. andR. D. Possee, The baculovirus expression system, Chapman and Hall, United Kingdom (1992); O’Reilly, D. R., L. K. Miller, V. A. Luckow, Baculovirus Expression Vectors: A Laboratory Manual, New York (1992); W. H. Freeman and Richardson, C D., Baculovirus Expression Protocols, Methods in Molecular Biology, volume 39 (1995),

[0162] In some embodiments, as a result of the infection of the host cell with a helper virus and / or an accessory function vector, a recombinant AAV particle is produced, and the produced rAAV particle is infectious, replication-defective virus, and includes an AAV protein shell that encapsulates a heterologous nucleotide sequence of interest flanked on both sides by AAV ITRs.

[0163] rAAV particles can be purified from the host cell using a purification method known in the art, such as chromatography, CsCl gradients, and other methods as described, for example, in U. S, Pat. Nos. 6,989,264 and 8,137,948 and WO 2010 / 148143. In some embodiments, residual helper virus can be inactivated using known methods, e.g., by heating.Pharmaceutical Compositions

[0164] In some embodiments, the present disclosure provides a pharmaceutical composition comprising the nucleic acid molecule, the vector, or the rAAV particle described herein. In some embodiments, the pharmaceutical composition is used in the prevention and / or treatment of a disease, disorder, or condition in a subject.

[0165] In some embodiments, the nucleic acid molecule, the vector, or the rAAV particle described herein are formulated as one or more pharmaceutical compositions. In some embodiments, the pharmaceutical composition comprises the nucleic acid molecule, the vector, or the rAAV particle described herein and a pharmaceutically acceptable carrier or excipient.

[0166] In some embodiments, viral particles comprising the nucleic acid molecules described herein are formulated as one or more pharmaceutical compositions. In some embodiments, the pharmaceutical composition comprises a viral particle comprising a viral vector or genome encoding a PC2 protein described herein and a pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition comprises an AAV particle comprising anAttorney Docket No. TORQ-012 / 02WO 339010-2071AAV vector or genome encoding a PC2 protein described herein and a pharmaceutically acceptable carrier or excipient.

[0167] A pharmaceutically acceptable carrier or excipient includes, without limitation, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, surfactant, and / or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans and / or domestic animals. Exemplary' pharmaceutically acceptable carriers include, but are not limited to, sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; tragacanth; malt; gelatin; talc; cocoa butter; waxes; animal and vegetable fats; paraffins; silicones; bentonites; silicic acid; zinc oxide; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; algimc acid; pyrogen- free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and any other compatible substances employed in pharmaceutical formulations. Except insofar as any conventional media and / or agent is incompatible with the agents of the present disclosure, its use in therapeutic compositions is contemplated. Supplementary active ingredients also can be incorporated into the compositions.

[0168] Pharmaceutically acceptable salt includes both acid and base addition salts. Pharmacally-acceptable salts include the acid addition salts (formed with the free ammo groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, algimc acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor- 10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonicAttorney Docket No. TORQ-012 / 02WO 339010-2071acid, mucic acid, naphthalene- 1,5-disulfonic acid, naphthal ene-2-sulfonic acid, 1 -hydroxyl-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, panioic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, ptoluenesulfomc acid, trifluoroacetic acid, undecylenic acid, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary' amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. In some embodiments, organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

[0169] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0170] Examples of pharmaceutically-acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha¬ tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0171] The pharmaceutical compositions provided herein can be provided as a controlled release or sustained release system. In some embodiments, a pump may be used to achieve controlled or sustained release (see, e.g., Sefton, Crit. Ref. Biomed. Eng. 14: 201-40 (1987); Buchwald et al., Surgery 88: 507-16 (1980); and Saudek et al., N. Engl. J. Med. 321: 569-74 (1989)). In some embodiments, polymeric materials can be used to achieve controlled or sustained release of aAttorney Docket No. TORQ-012 / 02WO 339010-2071prophylactic or therapeutic agent or a composition provided herein (see, e.g., Medical Applications of Controlled Release (Langer and Wise eds., 1974); Controlled Drug Bioavailability, Drug Product Design and Performance (Smolen and Ball eds., 1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23: 61-126 (1983); Levy et al., Science 228: 190-92 (1985); During et al., Ann. Neurol. 25: 351-56 (1989); Howard et al., J. Neurosurg. 71: 105-12 (1989); U. S. Pat Nos. 5,679,377; 5,916,597; 5,912,015; 5,989,463; and 5,128,326; PCT Publication Nos. WO 99 / 15154 and WO 99 / 20253). Examples of polymers used in sustained release formulations include, but are not limited to, poly (2-hydroxy ethyl methacrylate), poly (methyl methacrylate), poly (acrylic acid), poly (ethylene-co-vinyl acetate), poly (methacrylic acid), polyglycolides (PLG), polyanhydrides, poly (N-vinyl pyrrolidone), poly (vinyl alcohol), polyacrylamide, poly (ethylene glycol), polylactides (PL, A), poly (lactide-co-gly coll des) (PL, GA), and poly orthoesters. In some embodiments, the polymer used in a sustained release formulation is inert, free of leachable impurities, stable on storage, sterile, and biodegradable. In some embodiments, a controlled or sustained release system can be placed in proximity of a particular target tissue, for example, the kidney, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, Medical Applications of Controlled Release Vol. 2, 115-38 (1984)). Controlled release systems are discussed, for example, by Langer, Science 249: 1527-33 (1990). Any technique known to one of skill in the art can be used to produce sustained release formulations comprising one or more agents as described herein (see, e.g., U. S. Pat. No. 4,526,938, PCT publication Nos. WO 91 / 05548 and WO 96 / 20698, Ning et al., Radiotherapy & Oncology 39: 179-89 (1996); Song et al., PDA J. of Pharma. Sci. & Tech. 50: 372-97 (1995); Cleek et al., Pro. Int’l. Symp. Control. Rel. Bioact. Mater. 24: 853-54 (1997); and Lam et al.. Proc. IntT. Symp. Control Rel. Bioact. Mater. 24: 759-60 (1997)).10172] The active ingredients may also be entrapped in microcapsules prepared, for example, by coascervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacrylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington’s Pharmaceutical Sciences 18th edition.

[0173] In order for the pharmaceutical compositions to be used for in vivo administration, they are preferably sterile. The pharmaceutical composition may be rendered sterile by filtration throughAttorney Docket No. TORQ-012 / 02WO 339010-2071sterile filtration membranes. The pharmaceutical compositions provided herein generally can be placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0174] In some embodiments, the pharmaceutical compositions described herein are formulated in one of the following dosage forms: an intravenous dosage form, an intramuscular dosage form, an intraperitoneal dosage form, a subcutaneous dosage form, an oral dosage form, an intranasal dosage form, a suppository dosage form, an intradermal dosage form, a parenteral dosage form, a retro-ureteral dosage form, an intrathecal dosage form, a direct dosage form or a topical dosage form. In some embodiments, the pharmaceutical compositions described herein are formulated in an intravenous dosage form. In some embodiments, the pharmaceutical compositions described herein are formulated in a subcutaneous dosage form. In some embodiments, the pharmaceutical compositions described herein are formulated in an intramuscular dosage form. In some embodiments, the pharmaceutical compositions described herein are formulated in a retro-ureteral dosage form. In some embodiments, the pharmaceutical compositions described herein are formulated m an intravenous dosage form.

[0175] Further guidance regarding formulations that are suitable for various types of administration can be found in Remington’s Pharmaceutical Sciences, Mace Publishing Company, Philadelphia, Pa., 17th ed. (1985). For a brief review of methods for drug delivery, see, Langer, Science 249:1527-1533 (1990).Methods and Uses

[0176] In some embodiments, the nucleic acid molecule, the vector, or the rAAV particle described herein, or pharmaceutical composition thereof, are used in the prevention and / or treatment of a disease, disorder, or condition in a subject. In some embodiments, the disease or disorder is autosomal dominant polycystic kidney disease (ADPKD). In some embodiments, the disease or disorder is ADPKD type 2 (ADPKD2).

[0177] In some embodiments, the subject is human. In some embodiments, the subject is nonhuman primates (e.g., monkeys, baboons, and chimpanzees), mice, rats, bovines, horses, household cats, tigers and other large cats, dogs, pigs, rabbits, goats, deer, sheep, ferrets, gerbils, guinea pigs, hamsters, bats, and birds (e.g., chickens, turkeys, and ducks). In some embodiments, the subject may be a neonate, a juvenile, or an adult.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0178] In some embodiments, treating refers to the treatment of a disease in a mammal, e.g., in a human, including (a) inhibiting the disease, i.e., arresting disease development or preventing disease progression; (b) relieving the disease, i.e., causing regression of the disease state or relieving one or more symptoms of the disease; and (c) curing the disease, i.e., remission of one or more disease symptoms. In some embodiments, treatment results in an improvement or remediation of the symptoms of the disease. In some embodiments, treatment may refer to a short¬ term (e.g, temporary and / or acute) and / or a long-term (e.g., sustained) improvement or remediation in one or more disease symptoms. In some embodiments, the improvement is an observable or measurable improvement. In some embodiments, the improvement is an improvement m the general feeling of well-being of the subject. In some embodiments, the method inhibits or ameliorates renal cyst development,

[0179] In some embodiments, the nucleic acid molecule, the vector or rAAV particle described herein, or pharmaceutical composition thereof, is used in the prevention and / or treatment of a kidney disease. In some embodiments, the kidney disease is chronic kidney disease. In some embodiments, the kidney disease is end-stage renal disease. In some embodiments, the kidney disease is a genetic kidney disease. In some embodiments, the kidney disease is ADPKD. In some embodiments, the kidney disease is ADPKD1. In some embodiments, the kidney disease is ADPKD2 In some embodiments, the kidney disease is autosomal recessive PKD (ARPKD).

[0180] In some embodiments, the present disclosure provides a method of expressing PC2 protein in a subject, comprising administering an effective amount of the nucleic acid molecules of the present disclosure, the vectors or rAAV particles of the present disclosure, or pharmaceutical compositions of the present disclosure, wherein the effective amount is an amount that reduces at least one symptom of disease or condition in the subject. In some embodiments, the method inhibits or ameliorates renal cyst development in the subject. In some embodiments, the method reduces flank pain or back pain in the subject. In some embodiments, the method reduces abdominal enlargement in the subject. In some embodiments, the method reduces frequency of urinary tract infections in the subject. In some embodiments, the method reduces the number of kidney stones in the subject. In some embodiments, the method reduces the likelihood of kidney failure in the subject.

[0181] In some embodiments, the recombinant nucleic acid molecule encoding PC2 protein described herein, or pharmaceutical composition thereof, is delivered to a subject m need thereofAttorney Docket No. TORQ-012 / 02WO 339010-2071by methods known in the art. Such methods include, but are not limited to, transfection (e.g., lipid-mediated cationic polymers, calcium phosphate, dendrimers), electroporation or other methods of disrupting a membrane (e.g., nuclear transfection), viral delivery (e.g., lentivirus, retrovirus, adenovirus, AAV), microinjection, particle bombardment (“gene gun”), cell extrusion, light transfection, protoplast fusion, puncture infection, magnetic transfection, exosome-mediated transfer, lipid nanoparticle-mediated transfer, or any combination thereof. Administration of the recombinant nucleic acid molecule encoding PC2 protein described herein, or pharmaceutical composition thereof, can occur by infusion (e.g., continuous or bolus), injection, irrigation, inhalation, consumption, electro-osmosis, hemodialysis, iontophoresis, and other methods known in the art.

[0182] In some embodiments, the pharmaceutical compositions described herein are administered to subjects by one or more administration routes. Possible administration routes include, for example, intravenous, intramuscular, subcutaneous, intraperitoneal, oral, intranasal, intrasynovial, parenteral, intrapulmonary, transdermal, intrathecal, topical, retroureteral, and intralesional routes. In some embodiments, the administration route is selected from the group consisting of: intravenous administration, intramuscular administration, subcutaneous administration, intraperitoneal administration, retroureteral administration, and oral administration. In some embodiments, the pharmaceutical composition is administered intravenously to a subject (e.g., by intravenous infusion). In some embodiments, the pharmaceutical composition is administered by retrograde ureteral infusion. In some embodiments, the pharmaceutical composition is administered through the renal vein. In some embodiments, the pharmaceutical composition is administered through the renal artery. In some embodiments, the pharmaceutical composition is administered using a catheter. In some embodiments, the pharmaceutical composition is administered subcutaneously to a subject. In some embodiments, the pharmaceutical composition is administered retro-ureterally to a subject. In some embodiments, the pharmaceutical composition is administered intrathecally to a subject. In some embodiments, the pharmaceutical composition is administered intramuscularly to a subject. In some embodiments, administration route is direct, local, or systemic.

[0183] For prevention and treatment purposes, the pharmaceutical compositions described herein can be administered to a subject in a single bolus delivery, via continuous delivery (e.g., continuousAttorney Docket No. TORQ-012 / 02WO 339010-2071transdermal delivery') over an extended time period, or in a repeated administration protocol (e.g., on an hourly, daily, weekly, monthly, or yearly basis).

[0184] The effective amount of the pharmaceutical compositions administered to a particular subject will depend on a variety' of factors, several of which will differ from patient to patient including the disorder being treated and the severity of the disorder; activity of the specific agent(s) employed; the age, body weight, general health, sex and diet of the patient; the timing of administration, route of administration; the duration of the treatment; drugs used in combination; the judgment of the prescribing physician; and like factors known in the medical arts. Dosage amount and interval can be adjusted individually to provide plasma levels of the compound(s) which are sufficient to maintain therapeutic or prophylactic effect. In cases of local administration or selective uptake, such as local topical administration, the effective local concentration of active compound(s) cannot be related to plasma concentration. Skilled artisans will be able to optimize effective local dosages without undue experimentation.

[0185] In some embodiments, the vector or viral particle provided herein is administered at the concentration in vector genomes per milliliter (vg / mL) in the range of about 102vg / mL to about 1015vg / mL. In some embodiments, the concentration is in the range of 102vg / mL to about 1012vg / mL. In some embodiments, the concentration is in the range of 102vg / mL to about 1010vg / inL. In some embodiments, the concentration is in the range of 102vg / mL to about 105vg / mL. In some embodiments, the concentration is in the range of 105vg / mL to about 1010vg / mL. In some embodiments, the concentration is m the range of 105vg / mL to about 108vg / mL, In some embodiments, the concentration is in the range of 107vg / mL to about 1015vg / mL. In some embodiments, the concentration is in the range of 107vg / mL to about 1012vg / mL. In some embodiments, the concentration is in the range of 107vg / mL to about 1010vg / mL. In some embodiments, the concentration is in the range of 109vg / mL to about 1015vg / mL. In some embodiments, the concentration is in the range of 109vg / mL to about 1012vg / mL. In some embodiments, the concentration is in the range of 109vg / mL to about 1013vg / mL. In some embodiments, the concentration is in the range of 1011vg / mL to about 1013vg / mL. In some embodiments, the concentration is about 105vg / mL. In some embodiments, the concentration is about 106vg / mL. In some embodiments, the concentration is about 107vg / mL. In some embodiments, the concentration is about 108vg / mL. In some embodiments, the concentration is about 109vg / mL. In some embodiments, the concentration is about 1010vg / mL. In someAttorney Docket No. TORQ-012 / 02WO 339010-2071embodiments, the concentration is about 1011vg / mL. In some embodiments, the concentration is about 1012vg / mL. In some embodiments, the concentration is about 1013vg / mL. In some embodiments, the concentration is about 1014vg / mL In some embodiments, the concentration is about 1015vg / mL.

[0186] In some embodiments, the vector or viral particle provided herein is administered in a volume between about 0.1 mL and about 80 mL, for example between about 1 mL and about 80 mL, between about 10 mL and about 80 mL, between about 20 mL and about 80 mL, between about 40 mL and about 80 mL, between about 60 mL and about 80 mL, between about 80 mL and about 100 mL, between about 100 mL and about 200 mL, between about 200 mL and about 250 mL, between about 200 mL and about 300 mL, between about 300 mL and about 400 mL, between about 400 mL and about 500 mL, between about 0.3 mL and about 30 mL, between about 0.5 mL and about 30 mL, between about 1 mL and about 30 mL, between about 5 mL and about 30 mL, between about 0.1 mL and about 20 mL, between about 0.1 mL and about 10 mL, between about 0.1 mL and about 5.0 mL, between about 0.1 mL and about 2.0 mL, between about 0.1 mL and about 1.0 mL, between about 0.1 mL and about 0.8 mL, between about 0.1 mL and about 0.6 mL, between about 0.1 mL and about 0.4 mL, between about 0.1 mL and about 0.2 mL, between about 0.2 mL and about 1.0 mL, between about 0.2 mL and about 0.8 mL, between about 0.2 mL and about 0.6 mL, between about 0.2 mL and about 0.4 mL, between about 0.4 mL and about 1.0 mL, between about 0.4 mL and about 0.8 mL, between about 0.4 mL and about 0.6 mL, between about 0.6 mL and about 1.0 mL, between about 0.6 mL and about 0.8 mL, between about 0.8 mL and about 1.0 mL, or about 0.1 mL, about 0.2 mL, about 0.4 mL, about 0.6 mL, about 0.8 mL, about 1.0 mL, about 5.0 mL, about 10 mL, about 20 mL, about 30 mL, about 40 mL, about 50 mL, about 60 mL, about 70 mL, about 80 mL, about 90 mL, about 100 mL, about 125 mL, about 150 mL, about 175 mL, about 200 mL, about 225 mL, about 250 mL, about 275 mL, about 300 mL, about 325 mL, about 350 mL, about 375 mL, about 400 mL, about 425 mL, about 450 mL, about 475 mL, about 500 mL, or more.

[0187] In some embodiments, the vector or viral particle provided herein is administered to a subject at a dose of IxlO2to IxlO20vector genomes (vg). In some embodiments, the dose is about IxlO2to IxlO18vg. In some embodiments, the dose is about IxlO2to IxlO16vg. In some embodiments, the dose is about IxlO2to IxlO14vg. In some embodiments, the dose is about IxlO2to IxlO12vg. In some embodiments, the dose is about IxlO2to IxlO10vg. In some embodiments,Attorney Docket No. TORQ-012 / 02WO 339010-2071the dose is about IxlO2to IxlO5vg. In some embodiments, the dose is about IxlO5to IxlO12vg. In some embodiments, the dose is about IxlO5to 1x1010vg. In some embodiments, the dose is about IxlO5to IxlO8vg. In some embodiments, the dose is about IxlO7to IxlO18vg. In some embodiments, the dose is about IxlO7to IxlO16vg. In some embodiments, the dose is about 1x107to 1x1014vg. In some embodiments, the dose is about IxlO7to IxlO12vg. In some embodiments, the dose is about 1x107to 1x1010vg. In some embodiments, the dose is about 1x107to 1x108vg. In some embodiments, the dose is about IxlO9to IxlO18vg. In some embodiments, the dose is about IxlO9to IxlO16vg. In some embodiments, the dose is about IxlO9to IxlO14vg. In some embodiments, the dose is about IxlO9to IxlO12vg. In some embodiments, the dose is about IxlO9to IxlO10vg. In some embodiments, the dose is about IxlO11to IxlO18vg. In some embodiments, the dose is about 1x1011to 1x1016vg. In some embodiments, the dose is about 1x1011to 1x1014vg. In some embodiments, the dose is about IxlO11to IxlO12vg. In some embodiments, the dose is about IxlO13to IxlO18vg. In some embodiments, the dose is about IxlO13to IxlO16vg. In some embodiments, the dose is about IxlO13to IxlO14vg. In some embodiments, the dose is about IxlO15to IxlO18vg. In some embodiments, the dose is about IxlO15to IxlO16vg. In some embodiments, the dose is about lxl05vg. In some embodiments, the dose is about lxl06vg. In some embodiments, the dose is about lxl07vg. In some embodiments, the dose is about IxlO8vg. In some embodiments, the dose is about 1x109vg. In some embodiments, the dose is about IxlO10vg. In some embodiments, the dose is about IxlO11vg. In some embodiments, the dose is about lxl012vg. In some embodiments, the dose is about lxl013vg. In some embodiments, the dose is about lxl014vg. In some embodiments, the dose is about lxl015vg. In some embodiments, the dose is about lxl016vg. In some embodiments, the dose is about 1x1017vg. In some embodiments, the dose is about 1x1018vg. In some embodiments, the dose is about 1x1019vg. In some embodiments, the dose is about 1x1020vg. In some embodiments, the dose is the total dose. In some embodiments, the dose is for each administration.

[0188] In some embodiments, the vector or viral particle provided herein is administered to a subject at a dose of IxlO2to IxlO20vector genomes (vg) per kidney. In some embodiments, the dose is about IxlO2to IxlO18vg / kidney. In some embodiments, the dose is about IxlO2to IxlO16vg / kidney. In some embodiments, the dose is about IxlO2to IxlO14vg / kidney. In some embodiments, the dose is about IxlO2to IxlO12vg / kidney. In some embodiments, the dose is about IxlO2to IxlO10vg / kidney. In some embodiments, the dose is about IxlO2to IxlO5vg / kidney. InAttorney Docket No. TORQ-012 / 02WO 339010-2071some embodiments, the dose is about 1x105to 1x1012vg / kidney. In some embodiments, the dose is about IxlO3to IxlO10vg / kidney. In some embodiments, the dose is about IxlO5to IxlO8vg / kidney. In some embodiments, the dose is about IxlO7to IxlO18vg / kidney. In some embodiments, the dose is about IxlO7to IxlO16vg / kidney. In some embodiments, the dose is about IxlO7to IxlO14vg / kidney. In some embodiments, the dose is about IxlO7to IxlO12vg / kidney. In some embodiments, the dose is about 1x107to 1x1010vg / kidney. In some embodiments, the dose is about IxlO7to IxlO8vg / kidney. In some embodiments, the dose is about IxlO9to IxlO18vg / kidney. In some embodiments, the dose is about IxlO9to IxlO16vg / kidney. In some embodiments, the dose is about IxlO9to IxlO14vg / kidney. In some embodiments, the dose is about IxlO9to IxlO12vg / kidney. In some embodiments, the dose is about IxlO9to IxlO10vg / kidney. In some embodiments, the dose is about IxlO11to IxlO18vg / kidney. In some embodiments, the dose is about IxlO11to IxlO16vg / kidney. In some embodiments, the dose is about IxlO11to IxlO14vg / kidney. In some embodiments, the dose is about IxlO11to IxlO12vg / kidney. In some embodiments, the dose is about IxlO1-’ to IxlO18vg / kidney. In some embodiments, the dose is about IxlO13to IxlO16vg / kidney. In some embodiments, the dose is about IxlO13to IxlO14vg / kidney. In some embodiments, the dose is about IxlO15to IxlO18vg / kidney. In some embodiments, the dose is about 1x1015to 1x1016vg / kidney. In some embodiments, the dose is about IxlO5vg / kidney. In some embodiments, the dose is about 1x106vg / kidney. In some embodiments, the dose is about IxlO7vg / kidney. In some embodiments, the dose is about IxlO8vg / kidney. In some embodiments, the dose is about IxlO9vg / kidney. In some embodiments, the dose is about 1x1010vg / kidney. In some embodiments, the dose is about IxlO11vg / kidney. In some embodiments, the dose is about IxlO12vg / kidney. In some embodiments, the dose is about IxlO13vg / kidney. In some embodiments, the dose is about IxlO14vg / kidney. In some embodiments, the dose is about IxlO15vg / kidney. In some embodiments, the dose is about IxlO16vg / kidney. In some embodiments, the dose is about 1x1017vg / kidney. In some embodiments, the dose is about IxlO18vg / kidney. In some embodiments, the dose is about IxlO19vg / kidney. In some embodiments, the dose is about IxlO20vg / kidney. In some embodiments, the dose is the total dose. In some embodiments, the dose is for each administration.

[0189] In some embodiments, the vector or viral particle provided herein is administered to a subject at a dose of IxlO2to IxlO20vector genomes per kilogram (vg / kg) body weight. In some embodiments, the dose is about IxlO2to IxlO18vg / kg body weight. In some embodiments, theAttorney Docket No. TORQ-012 / 02WO 339010-2071dose is about 1x102to 1x1016vg / kg. In some embodiments, the dose is about IxlO2to IxlO14vg / kg body weight. In some embodiments, the dose is about 1x102to 1x1012vg / kg body weight. In some embodiments, the dose is about IxlO2to IxlO10vg / kg body weight. In some embodiments, the dose is about IxlO2to IxlO5vg / kg body weight. In some embodiments, the dose is about IxlO5to IxlO12vg / kg body weight. In some embodiments, the dose is about IxlO5to IxlO10vg / kg body weight. In some embodiments, the dose is about 1x105to 1x108vg / kg body weight. In some embodiments, the dose is about IxlO7to IxlO18vg / kg body weight. In some embodiments, the dose is about 1x10' to IxlO16vg / kg body weight. In some embodiments, the dose is about 1x107to 1x1014vg / kg body weight. In some embodiments, the dose is about IxlO7to IxlO12vg / kg body weight. In some embodiments, the dose is about IxlO7to IxlO10vg / kg body weight. In some embodiments, the dose is about IxlO7to IxlO8vg / kg body weight. In some embodiments, the dose is about IxlO9to IxlO18vg / kg body weight. In some embodiments, the dose is about IxlO9to IxlO16vg / kg body weight. In some embodiments, the dose is about IxlO9to IxlO14vg / kg body weight. In some embodiments, the dose is about IxlO9to IxlO12vg / kg body weight. In some embodiments, the dose is about IxlO9to IxlO10vg / kg body weight. In some embodiments, the dose is about IxlO11to 1x1018vg / kg body weight. In some embodiments, the dose is about IxlO11to IxlO16vg / kg body weight. In some embodiments, the dose is about 1x1011to IxlO14vg / kg body weight. In some embodiments, the dose is about IxlO11to IxlO12vg / kg body weight. In some embodiments, the dose is about IxlO13to IxlO18vg / kg body weight. In some embodiments, the dose is about IxlO13to IxlO16vg / kg body weight. In some embodiments, the dose is about IxlO13to IxlO14vg / kg body weight. In some embodiments, the dose is about IxlO15to IxlO18vg / kg body weight. In some embodiments, the dose is about 1x1015to 1x1016vg / kg body weight. In some embodiments, the dose is about IxlO5vg / kg body weight. In some embodiments, the dose is about IxlO6vg / kg body weight. In some embodiments, the dose is about IxlO7vg / kg body weight. In some embodiments, the dose is about 1x108vg / kg body weight. In some embodiments, the dose is about 1x109vg / kg body weight. In some embodiments, the dose is about IxlO10vg / kg body weight. In some embodiments, the dose is about IxlO11vg / kg body weight. In some embodiments, the dose is about IxlO12vg / kg body weight. In some embodiments, the dose is about IxlO13vg / kg body weight. In some embodiments, the dose is about IxlO14vg / kg body weight. In some embodiments, the dose is about IxlO15vg / kg body weight. In some embodiments, the dose is about 1x1016vg / kg body weight. In some embodiments, the dose is about IxlO17vg / kg body weight. InAttorney Docket No. TORQ-012 / 02WO 339010-2071some embodiments, the dose is about 1x1018vg / kg body weight. In some embodiments, the dose is about 1x1019vg / kg body weight. In some embodiments, the dose is about 1x1020vg / kg body weight. In some embodiments, the dose is the total dose. In some embodiments, the dose is for each administration.

[0190] Dosages may be administered in single or multiple administrations, including, e.g., multiple weekly, bi-weekly, monthly, or yearly administrations. In some embodiments, a single dose of the pharmaceutical composition comprising the circular RNA is administered to a subject in need thereof. In some embodiments, the subject receives two or more doses of the pharmaceutical composition comprising the nucleic acid molecule encoding PC2 protein. In some embodiments, the subject receives two or more doses of the pharmaceutical composition comprising the nucleic acid molecule encoding PC2 protein, wherein consecutive doses are separated by at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, or at least a week. In some embodiments, the subject receives two or more doses of the pharmaceutical composition comprising the nucleic acid molecule encoding PC2 protein, wherein consecutive doses are separated by a period of at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, at least six weeks, at least seven weeks, at least eight weeks, at least one month, at least two months, or at least three months. In some embodiments, two or more doses of the pharmaceutical composition comprising the nucleic acid molecule encoding PC2 protein are administered to a subject in need thereof separated by a period of about one week to about two weeks, about two weeks to about four weeks, about one month to about two months, about two months to about four months, or about one month to about six months. In some embodiments, the duration of the administration of the pharmaceutical composition comprising the nucleic acid molecule encoding PC2 protein is at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least one week, at least two weeks, at least three weeks, at least one month, at least two months, at least three months, at least four months, at least five months, at least six months, at least nine months, at least one year, at least two years, at least three years, at least four years, or at least five years. In some embodiments, administration of the pharmaceutical composition comprising the nucleic acid molecule encoding PC2 protein is on an irregular basis as indicated by monitoring clinical symptoms of the disease in a subject in need thereof. In some embodiments, administration of the pharmaceutical composition comprising the nucleic acid molecule encoding PC2 protein is up to the lifetime of a subject in need thereof. TheAttorney Docket No. TORQ-012 / 02WO 339010-2071exact protocols depend upon the disease or condition, the stage of the disease and parameters of the individual subject being treated.

[0191] In some embodiments, the nucleic acid molecules, vectors, or rAAV particles disclosed herein, or pharmaceutical compositions thereof are administered in combination with additional therapeutic composition(s). In some embodiments, the pharmaceutical compositions disclosed herein and the additional therapeutic composition(s) are administered simultaneously. In some embodiments, the pharmaceutical compositions disclosed herein are administered before the additional therapeutic composition(s). In some embodiments, the pharmaceutical compositions disclosed herein are administered after the additional therapeutic composition(s). The additional therapeutic compositions include, but are not limited to, an inhibitor of a vasopressin receptor (e.g., tolvaptan), angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers (ARBs), pain relievers (e.g., acetaminophen), antibiotics, pasireotide, and anti-miR-17 oligonucleotide RGLS4326.

[0192] In some embodiments, PKD2 mRNA is expressed in a cell or tissue of the subject following administration of the pharmaceutical composition. In some embodiments, the expression of PKD2 mRNA is increased in a cell or tissue of the subject compared to baseline (i.e., before treatment with the pharmaceutical composition comprising the nucleic acid molecules described herein). In some embodiments, the expression of PKD2 mRNA is increased by at least about 2-fold, about 5- fold, about 10-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 60-fold, about 70-fold, about 80-fold, about 90-fold, about 100-fold, about 500-fold, about 1000-fold in a cell or tissue of the subject compared to baseline.

[0193] In some embodiments, PC2 protein is expressed in a cell or tissue of the subject following administration of the pharmaceutical composition. In some embodiments, the expression of PC2 protein is increased in a cell or tissue of the subject compared to baseline (i.e., before treatment with the pharmaceutical composition comprising the nucleic acid molecules described herein). In some embodiments, the expression of PC2 protein is increased by at least about 2-fold, about 5-fold, about 10-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 60-fold, about 70-fold, about 80-fold, about 90-fold, about 100-fold, about 500-fold, about 1000-fold in a cell or tissue of the subject compared to baseline.Attorney Docket No. TORQ-012 / 02WO 339010-2071Kits and Articles of Manufacture

[0194] Also provided herein are kits or articles of manufacture for carrying out methods described herein. At least one component of the kit or article of manufacture is the nucleic acid molecules disclosed in Section 6.2 above, the vectors or rAAV particles disclosed in Section 6.3 above, or the pharmaceutical compositions disclosed in Section 6.4 above.

[0195] In some embodiments, the kit or article of manufacture comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, balloon occlusion ureteral catheter, in-line pressure transducer, stopcock, extension tubing, syringe, syringe infusion pump, and guidewire, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds a composition which is by itself or combined with another composition effective for treating, preventing and / or diagnosing the condition and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper that is pierceable by a hypodermic injection needle).

[0196] In some embodiments, the kit or article of manufacture comprises a cystoscope, a guidewire, a balloon occlusion catheter, an in-line pressure transducer, a pressure wire, a stopcock and extension tubing, syringe, peristaltic pump, and / or syringe pump.

[0197] In some embodiments, the label or package insert indicates that at least one component of the kit or article of manufacture is used for treating the disorder of choice. In some embodiments, the label or package insert comprises instructions for practicing the methods provided herein. The instructions are generally recorded on a suitable recording medium. In some embodiments, the instructions may be printed on a substrate, such as paper or plastic, etc. In some embodiments, the instructions are present as an electronic storage data file present on a suitable computer readable storage medium, e.g., CD-ROM, diskette, flash drive, etc. In some embodiments, the actual instructions are not present in the kit, but means for obtaining the instructions from a remote source, e.g., via the internet, are provided. An example of this embodiment is a kit or article of manufacture that includes a web address where the instructions can be viewed and / or from which the instructions can be downloaded.

[0198] In some embodiments, the kit or article of manufacturer further comprises other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.Attorney Docket No. TORQ-012 / 02WO 339010-2071NUMBERED EMBODIMENTS

[0199] Notwithstanding the appended claims, the disclosure sets forth the following embodiments:

[0200] Embodiment 1. A nucleic acid molecule comprising a polynucleotide encoding a polycystin- 2 (PC2) protein, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 3-5 and 7-9.

[0201] Embodiment 2. A nucleic acid molecule comprising a polynucleotide encoding a polycystin- 2 (PC2) protein, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 5 or SEQ ID NO: 9.

[0202] Embodiment 3. The nucleic acid molecule of embodiment 1 or 2, wherein the PC2 protein comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 1.

[0203] Embodiment 4 The nucleic acid molecule of any one of embodiments 1-3, wherein the PC2 protein comprises an amino acid sequence of SEQ ID NO: 1.

[0204] Embodiment 5. The nucleic acid molecule of any one of embodiments 1-4, wherein the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 3-5 and 7-9.

[0205] Embodiment 6. The nucleic acid molecule of any one of embodiments 1-5, wherein the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 5 or SEQ ID NO: 9.

[0206] Embodiment 7. The nucleic acid molecule of any one of embodiments 1-6, wherein the polynucleotide is operably linked to a promoter.

[0207] Embodiment 8. The nucleic acid molecule of embodiment 7, wherein the promoter is selected from the group consisting of: chicken beta actin (CBA) promoter, a human phosphoglycerate kinase (hPGK) promoter, a human glucose-6-phosphatase (hG6Pase) promoter, a mouse polycystic kidney disease 2 (mPKD2) promoter, a mouse E-cadherin promoter (mECAD), a mouse kidney-specific cadherin (mKSPC) promoter, a mouse paired box gene 8 (mPax8) promoter, a human with-no-lysine kinase 1 promoter with a renal enhancer element (hWNKl-Attorney Docket No. TORQ-012 / 02WO 339010-2071RP), a human phosphoglycerate kinase promoter with renal enhancer element (RP-hPGK), and a human truncated PKD2 promoter, a pig truncated PKD2 promoter, and a mouse truncated PKD2 promoter.

[0208] Embodiment 9. The nucleic acid molecule of embodiment 7 or 8, wherein the promoter is a CBA promoter.

[0209] Embodiment 10. The nucleic acid molecule of embodiment 7 or 8, wherein the promoter is an hPGK promoter.

[0210] Embodiment 11. The nucleic acid molecule of any one of embodiments 7-10, wherein the promoter comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 12-15 and 21-32.

[0211] Embodiment 12. The nucleic acid molecule of any one of embodiments 7-11, wherein the promoter comprises a nucleic acid sequence of any one of SEQ ID NOs: 12-15 and 21-32.

[0212] Embodiment 13. The nucleic acid molecule of any one of embodiments 7-9, 11, and 12, wherein the promoter comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 12.

[0213] Embodiment 14. The nucleic acid molecule of any one of embodiments 7-9, 11, 12, and 13 wherein the promoter comprises a nucleic acid sequence of SEQ ID NO: 12.

[0214] Embodiment 15. The nucleic acid molecule of any one of embodiments 7, 8, and 10-12, wherein the promoter comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 13.

[0215] Embodiment 16. The nucleic acid molecule of any one of embodiments 7, 8, 10-12, and 15, wherein the promoter comprises a nucleic acid sequence of SEQ ID NO: 13.

[0216] Embodiment 17. The nucleic acid molecule of any one of embodiments 1-16, wherein the polynucleotide is operably linked to a polyadenylation signal.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0217] Embodiment 18. The nucleic acid molecule of embodiment 17, wherein the polyadenylation signal is a bovine growth hormone poly A (bGHpA) signal.

[0218] Embodiment 19. The nucleic acid molecule of embodiment 17 or 18, wherein the polyadenylation signal comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 18.

[0219] Embodiment 20. The nucleic acid molecule of any one of embodiments 17-19, wherein the polyadenylation signal comprises a nucleic acid sequence of SEQ ID NO: 18.

[0220] Embodiment 21. The nucleic acid molecule of embodiment 17, wherein the polyadenylation signal is a rabbit [3-globin polyadenylation (rBGHpA) signal.

[0221] Embodiment 22. The nucleic acid molecule of embodiment 17 or 21, wherein the polyadenylation signal comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91 %, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 33.

[0222] Embodiment 23. The nucleic acid molecule of embodiment 17, 21, or 22, wherein the polyadenylation signal comprises a nucleic acid sequence of SEQ ID NO: 33.

[0223] Embodiment 24. The nucleic acid molecule of any one of embodiments 1-23, wherein the polynucleotide is operably linked to a 5’ untranslated region (UTR).

[0224] Embodiment 25. The nucleic acid molecule of embodiment 24, wherein the 5’ U TR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 34-49.

[0225] Embodiment 26. The nucleic acid molecule of embodiment 24 or 25, wherein the 5’ UTR comprises a nucleic acid sequence of any one of SEQ ID NOs: 34-49.

[0226] Embodiment 27. The nucleic acid molecule of any one of embodiments 24-26, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, atAttorney Docket No. TORQ-012 / 02WO 339010-2071least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 35.

[0227] Embodiment 28. The nucleic acid molecule of any one of embodiments 24-27, wherein the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 35.

[0228] Embodiment 29. The nucleic acid molecule of any one of embodiments 1-28, wherein the polynucleotide is operably linked to a 3’ UTR.

[0229] Embodiment 30. The nucleic acid molecule of embodiment 29, wherein the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 50-61.

[0230] Embodiment 31. The nucleic acid molecule of embodiment 29 or 30, wherein the 3’ UTR comprises a nucleic acid sequence of any one of SEQ ID NOs: 50-61.

[0231] Embodiment 32. The nucleic acid molecule of any one of embodiments 29-31, wherein the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 58.

[0232] Embodiment 33. The nucleic acid molecule of any one of embodiments 29-32, wherein the 3’ UTR comprises a nucleic acid sequence of SEQ ID NO: 58.

[0233] Embodiment 34. The nucleic acid molecule of any one of embodiments 1-23, wherein the polynucleotide is operably linked to a 5’ LTTR and a 3’ UTR.

[0234] Embodiment 35. The nucleic acid molecule of embodiment 34, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any one of SEQ ID NOs: 34-49; and the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any one of SEQ ID NOs: 50-61.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0235] Embodiment 36. The nucleic acid molecule of embodiment 34 or 35, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 44; and the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 50.

[0236] Embodiment 37. The nucleic acid molecule of embodiment 34 or 35, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 37; and the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 61.

[0237] Embodiment 38 The nucleic acid molecule of any one of embodiments 1-37, wherein the polynucleotide comprises a translation initiation sequence.

[0238] Embodiment 39. The nucleic acid molecule of embodiment 38, wherein the translation initiation sequence comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 20.

[0239] Embodiment 40. The nucleic acid molecule of embodiment 38 or 39, wherein the translation initiation sequence comprises a nucleic acid sequence of SEQ ID NO: 20.

[0240] Embodiment 41. The nucleic acid molecule of any one of embodiments 1-40, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 108-155.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0241] Embodiment 42. The nucleic acid molecule of any one of embodiments 1-41, wherein the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 108-155.

[0242] Embodiment 43. The nucleic acid molecule of any one of embodiments 1 -40, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 109 or SEQ ID NO: 153.

[0243] Embodiment 44. The nucleic acid molecule of any one of embodiments 1-41, wherein the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 109 or SEQ ID NO: 153,

[0244] Embodiment 45. The nucleic acid molecule of any one of embodiments 1 -40, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 110 or SEQ ID NO: 154.

[0245] Embodiment 46. The nucleic acid molecule of any one of embodiments 1-41, wherein the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 110 or SEQ ID NO: 154.

[0246] Embodiment 47. A vector comprising the nucleic acid molecule of any one of embodiments 1-46.

[0247] Embodiment 48. The vector of embodiment 47, wherein the vector is a viral vector.

[0248] Embodiment 49. The vector of embodiment 48, wherein the viral vector is a recombinant adeno-associated virus (rAAV) vector.

[0249] Embodiment 50. The vector of embodiment 49, wherein the rAAV vector comprises a 5’ inverted terminal repeat (ITR) and a 3’ ITR, wherein the 5’ ITR flanks at the 5’ end of the nucleic acid molecule and the 3’ ITR flanks at the 3’ end of the nucleic acid molecule.

[0250] Embodiment 51. The vector of embodiment 50, wherein the 5’ ITR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 16.

[0251] Embodiment 52. The vector of embodiment 50 or 51, wherein the 5’ ITR comprises a nucleic acid sequence of SEQ ID NO: 16.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0252] Embodiment 53. The vector of any one of embodiments 50-52, wherein the 3’ ITR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 17.

[0253] Embodiment 54. The vector of any one of embodiments 50-53, wherein the 3’ ITR comprises a nucleic acid sequence of SEQ ID NO: 17.

[0254] Embodiment 55. The vector of any one of embodiments 47-54, wherein the rAAV vector is derived from AAV1, AAV2, AAV2i8, AAV3, AAV3-B, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrhS, AAVrh8R, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAV-DJ, AAV LK03, AAVKP1, AAV.cc47, AAV.cc84, AAVrh74, AAV44-9, or a variant thereof.

[0255] Embodiment 56. The vector of any one of embodiments 47-55, wherein the rAAV vector comprises a polynucleotide sequence that is at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 65-107 and 156-160.

[0256] Embodiment 57. The vector of any one of embodiments 47-56, wherein the rAAV vector comprises a polynucleotide sequence of any one of SEQ ID NOs: 65-107 and 156-160.

[0257] Embodiment 58. The vector of any one of embodiments 47-57, wherein the rAAV vector comprises a polynucleotide sequence that is at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 66 or SEQ ID NO: 158.

[0258] Embodiment 59. The vector of any one of embodiments 47-58, wherein the rAAV vector comprises a polynucleotide sequence of SEQ ID NO: 66 or SEQ ID NO: 158.

[0259] Embodiment 60. The vector of any one of embodiments 47-57, wherein the rAAV vector comprises a polynucleotide sequence that is at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 67 or SEQ ID NO: 159.

[0260] Embodiment 61. The vector of any one of embodiments 47-57 and 60, wherein the rAAV vector comprises a polynucleotide sequence of SEQ ID NO: 67 or SEQ ID NO: 159.

[0261] Embodiment 62. A rAAV particle comprising:Attorney Docket No. TORQ-012 / 02WO 339010-2071(a) the nucleic acid molecule of any one of embodiments 1-46, or the vector of any one of embodiments 47 to 61; and(b) a capsid of AAV1, AAV2, AAV2i8, AAV3, AAV3-B, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh8R, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAV-DJ, AAV LK03, AAV KP1, AAV.cc47, AAV.cc84, AAVrh74, AAV44-9, or a variant thereof.

[0262] Embodiment 63. The rAAV particle of embodiment 62, wherein the rAAV particle transduces kidney cells.

[0263] Embodiment 64. The rAAV particle of embodiment 62 or 63, wherein the rAAV particle transduces distal tubule cells, proximal tubule cells, and / or collecting duct cells.

[0264] Embodiment 65 The rAAV particle of any one of embodiments 62-64, wherein the capsid protein is an AAV9 capsid variant.

[0265] Embodiment 66. The AAV particle of any one of embodiments 62-65, wherein the capsid protein is an AAV.kl 3 capsid protein.

[0266] Embodiment 67. The AAV particle of embodiment 66, wherein the AAV.kl 3 capsid protein comprises an amino acid sequence of SEQ ID NO: 63 with 1, 2, 3, 4, or more ammo acid substitutions at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

[0267] Embodiment 68 The AAV particle of embodiment 66 or 67, wherein the AAV.kl 3 capsid protein comprises an amino acid sequence of SEQ ID NO: 63 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

[0268] Embodiment 69. The AAV particle of any one of embodiments 66-68, wherein the AAV.kl 3 capsid protein consists of an amino acid sequence of SEQ ID NO: 63 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

[0269] Embodiment 70. The AAV particle of any one of embodiments 66-69, wherein the AAV.kl 3 capsid increases transduction of kidney cells by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, or more, compared to a wild-type AAV9 capsid.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0270] Embodiment 71. The AAV particle of any one of embodiments 62-65, wherein the capsid protein is an AAV.k20 capsid protein.

[0271] Embodiment 72. The AAV particle of embodiment 71, wherein the AAV.k20 capsid protein comprises an amino acid sequence of SEQ ID NO: 64 with 1, 2, 3, 4, or more amino acid substitutions at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

[0272] Embodiment 73. The AAV particle of embodiment 71 or 72, wherein the AAV.k20 capsid protein comprises an amino acid sequence of SEQ ID NO: 64 at positions 452-458, wherein positions 452-458 of the AA V capsid protein are numbered with reference to SEQ ID NO: 62,

[0273] Embodiment 74, The AAV particle of any one of embodiments 71-73, wherein the AAV.k20 capsid protein consists of an amino acid sequence of SEQ ID NO: 64 at positions 452-458, wherein positions 452-458 of the A AV capsid protein are numbered with reference to SEQ ID NO: 62.

[0274] Embodiment 75, The AAV particle of any one of embodiments 71-74, wherein the AAV.k20 capsid increases transduction of kidney cells by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, or more, compared to a wild-type AAV9 capsid.

[0275] Embodiment 76. A pharmaceutical composition, comprising the nucleic acid molecule of any one of embodiments 1-46, the vector of any one of embodiments 47-61, or the rAAV particle of any one of embodiments 62-75, and a pharmaceutically acceptable excipient.

[0276] Embodiment 77. A method of treating a disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of the nucleic acid molecule of any one of embodiments 1-46, the vector of any one of embodiments 47-61, the rAAV particle of any one of embodiments 62-75, or the pharmaceutical composition of embodiment 76.

[0277] Embodiment 78. The method of embodiment 77, wherein the nucleic acid molecule of any one of embodiments 1-46, the vector of any one of embodiments 47-61, the rAAV particle of any one of embodiments 62-75, or the pharmaceutical composition of embodiment 76 is administered via retrograde ureteral infusion.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0278] Embodiment 79. The method of embodiment 77, wherein the nucleic acid molecule of any one of embodiments 1-46, the vector of any one of embodiments 47-61, the rAAV particle of any one of embodiments 62-75, or the pharmaceutical composition of embodiment 76 is administered via the arterial route.

[0279] Embodiment 80. The method of any one of embodiments 77-79, wherein the nucleic acid molecule of any one of embodiments 1-46, the vector of any one of embodiments 47-61, the rAAV particle of any one of embodiments 62-75, or the pharmaceutical composition of embodiment 76 is administered at a dose of about 1E11 vector genomes per kidney (vg / kidney) to about 1E16 vg / kidney.

[0280] Embodiment 81. The method of any one of embodiments 77-79, wherein the nucleic acid molecule of any one of embodiments 1-46, the vector of any one of embodiments 47-61, the rAAV particle of any one of embodiments 62-75, or the pharmaceutical composition of embodiment 76 is administered at a dose of about 1E11 vector genomes per kilogram (vg / kg) body weight to about 1E15 vg / kg body weight

[0281] Embodiment 82. The method of any one of embodiments 77-81, wherein the disease or disorder is autosomal dominant polycystic kidney disease (ADPKD).

[0282] Embodiment 83. The method of any one of embodiments 77-82, wherein the disease or disorder is ADPKD type 2 (ADPKD2)

[0283] Embodiment 84. The method of any one of embodiments 77-83, wherein the method inhibits, ameliorates, or reverses renal cyst development.

[0284] Embodiment 85. The method of any one of embodiments 77-84, wherein the method inhibits, ameliorates, or reverses decline in kidney function.EXAMPLES

[0285] The following is a description of various methods and materials used in the studies. They are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments below were performed and are all of the experiments that may be performed. It is to be understood that exemplary descriptions written in the present tense were not necessarily performed, but rather that the descriptions can be performed to generate the data and the likeAttorney Docket No. TORQ-012 / 02WO 339010-2071associated with the teachings of the present invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, percentages, etc.), but some experimental errors and deviations should be accounted for.Example 1 — Design of PKD2 Cassettes for Expressing PC2

[0286] PKD2 expression cassettes were designed as shown in Fig. 1. Each PKD2 cassette comprised one of four promoters combined with one of four PKD2 sequences encoding polycystin-2 protein (PC2). Promoters included chicken beta actin (CBA), human phosphoglycerate kinase (hPGK), human glucose-6-phosphatase (hG6Pase), and mouse polycystic kidney disease 2 (mPKD2). PKD2 coding sequences included unmodified endogenous full-length PKD2 (PKD2-FL), codon-optimized PKD2 (PKD2-FL; CO), CpG-depleted PKD2 (PKD2-FL; ACpG), and a version of PKD2 that was both codon-optimized and CpG-depleted (PKD2-FL; CO; ACpG). All cassettes were terminated with bovine growth hormone poly A (bGHpA) signal, and the entire promoter-coding sequence-poly A cassette was flanked by wild-type AAV2 inverted terminal repeats (5’ and 3’ ITRs). All PKD2 coding sequences comprised a Kozak sequence at the 5’ end and were synthesized with and without a sequence encoding a C-terminal linker and 3 repeating HA-tags (3X-HA).

[0287] Sequences related to the PKD2 cassettes are listed in Table 3 below.Table 3. Sequences for PKD2 Cassette DesignDescription Amino Acid or Nucleotide Sequence SEQ ID NOHuman PC2 MVNSSRVQPQQPGDAKRPPAPRAPDPGRLMAGCAAVGASLAAPGG 1 LCEQRGLEIEMQRIRQAAARDPPAGAAASPSPPLSSCSRQAWSRDNP GFEAEEEEEEVEGEEGGMVVEMDVEWRPGSRRSAASSAVSSVGARS RGLGGYHGAGHPSGRRRRREDQGPPCPSPVGGGDPLHRHLPLEGQP PRVAWAERLVRGLRGLWGTRLMEESSTNREKYLKSVLRELVTYLLF LIVLCILTYGMMSSNVYYYTRMMSQLFLDTPVSKTEKTNFKTLSSME DFWKFTEGSLLDGLYWKMQPSNQTEADNRSFIFYENLLLGVPRIRQL RVRNGSCSIPQDLRDEIKECYDVYSVSSEDRAPFGPRNGTAWIYTSEK DLNGSSHWGIIATYSGAGYYLDLSRTREETAAQVASLKKNWLDRG TRATFIDFSVYNANINLFCVVRLLVEFPATGGVIPSWQFQPLKLIRYV TTFDFFLAACEIIFCFFIFYYWEEILEIRIHKLHYFRSFWNCLDVVIW LSVVAIGINIYRTSNVEVLLQFLEDQNTFPNFEHLAYWQIQFNNIAAV TVFFVWIKLFKFINFNRTMSQLSTTMSRCAKDLFGFAIMFFIIFLAYA QLAYLVFGTQVDDFSTFQECIFTQFRIILGDINFAEIEEANRVLGPIYFT TFVFFMFFILLNMFLAIINDTYSEVKSDLAQQKAEMELSDLIRKGYHK ALVKLKLKKNTVDDISESLRQGGGKLNFDELRQDLKGKGHTDAEIEAIFTKYDQDGDQELTEHEHQQMRDDLEKEREDLDLDHSSLPRPMSSAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO RSFPRSLDDSEEDDDEDSGHSSRRRGSISSGVSYEEFQVLVRRVDRM EHSIGSIVSKIDAVIVKLEIMERAKLKRREVLGRLLDGVAEDERLGRD SEIHREQMERLVREELERWESDDAASQISHGLGTPVGLNGQPRPRSS RPSSSQSTEGMEGAGGNGSSNVHVHuman PKD2: ATGGTGAACTCCAGTCGCGTGCAGCCTCAGCAGCCCGGGGACGC 2 Full Length CAAGCGGCCGCCCGCGCCCCGCGCGCCGGACCCGGGCCGGCTGA TGGCTGGCTGCGCGGCCGTGGGCGCCAGCCTCGCCGCCCCGGGCG GCCTCTGCGAGCAGCGGGGCCTGGAGATCGAGATGCAGCGCATC CGGCAGGCGGCCGCGCGGGACCCCCCGGCCGGAGCCGCGGCCTC CCCTTCTCCTCCGCTCTCGTCGTGCTCCCGGCAGGCGTGGAGCCG CGATAACCCCGGCTTCGAGGCCGAGGAGGAGGAGGAGGAGGTGG AAGGGGAAGAAGGCGGAAT GGT GGT GGAGAT GGACGTAGAGT G GCGCCCGGGCAGCCGGAGGTCGGCCGCCTCCTCGGCCGTGAGCTC CGTGGGCGCGCGGAGCCGGGGGCTTGGGGGCTACCACGGCGCGG GCCACCCGAGCGGGAGGCGGCGCCGGCGAGAGGACCAGGGCCCG CCGTGCCCCAGCCCAGTCGGCGGCGGGGACCCGCTGCATCGCCAC CTCCCCCTGGAAGGGCAGCCGCCCCGAGTGGCCTGGGCGGAGAG GCTGGTTCGCGGGCTGCGAGGTCTCTGGGGAACAAGACTCATGG AGGAAAGCAGCACTAACCGAGAGAAATACCTTAAAAGTGTTTTA CGGGAACTGGTCACATACCTCCTTTTTCTCATAGTCTTGTGCATCT TGACCTACGGCATGATGAGCTCCAATGTGTACTACTACACCCGGA TGATGTCACAGCTCTTCCTAGACACCCCCGTGTCCAAAACGGAGA AAACTAACTTTAAAACTCTGTCTTCCATGGAAGACTTCTGGAAGT TCACAGAAGGCTCCTTATTGGATGGGCTGTACTGGAAGATGCAGC CCAGCAACCAGACTGAAGCTGACAACCGAAGTTTCATCTTCTATG AGAACCTGCTGTTAGGGGTTCCACGAATACGGCAACTCCGAGTCA GAAATGGATCCTGCTCTATCCCCCAGGACTTGAGAGATGAAATTA AAGAGTGCTATGATGTCTACTCTGTCAGTAGTGAAGATAGGGCTC CCTTTGGGCCCCGAAATGGAACCGCTTGGATCTACACAAGTGAAA AAGACTTGAATGGTAGTAGCCACTGGGGAATCATTGCAACTTATA GTGGAGCTGGCTATTATCTGGATTTGTCAAGAACAAGAGAGGAA ACAGCTGCACAAGTTGCTAGCCTCAAGAAAAATGTCTGGCTGGAC CGAGGAACCAGGGCAACTTTTATTGACTTCTCAGTGTACAACGCC AACATTAACCTGTTCTGTGTGGTCAGGTTATTGGTTGAATTCCCAG CAACAGGTGGTGTGATTCCATCTTGGCAATTTCAGCCTTTAAAGC TGATCCGATATGTCACAACTTTTGATTTCTTCCTGGCAGCCTGTGA GATTATCTTTTGTTTCTTTATCTTTTACTATGTGGTGGAAGAGATA TTGGAAATTCGCATTCACAAACTACACTATTTCAGGAGTTTCTGG AATTGTCTGGATGTTGTGATCGTTGTGCTGTCAGTGGTAGCTATA GGAATTAACATATACAGAACATCAAATGTGGAGGTGCTACTACA GTTTCTGGAAGATCAAAATACTTTCCCCAACTTTGAGCATCTGGC ATATTGGCAGATACAGTTCAACAATATAGCTGCTGTCACAGTATT TTTTGTCTGGATTAAGCTCTTCAAATTCATCAATTTTAACAGGACC ATGAGCCAGCTCTCGACAACCATGTCTCGATGTGCCAAAGACCTG TTTGGCTTTGCTATTATGTTCTTCATTATTTTCCTAGCGTATGCTCA GTTGGCATACCTTGTCTTTGGCACTCAGGTCGATGACTTCAGTACT TTCCAAGAGTGTATCTTCACTCAATTCCGTATCATTTTGGGCGATATCAACTTTGCAGAGATTGAGGAAGCTAATCGAGTTTTGGGACCAAAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO TTTATTTCACTACATTTGTGTTCTTTATGTTCTTCATTCTTTTGAAT ATGTTTTTGGCTATCATCAATGATACTTACTCTGAAGTGAAATCTG ACTTGGCACAGCAGAAAGCTGAAATGGAACTCTCAGATCTTATCA GAAAGGGCTACCATAAAGCTTTGGTCAAACTAAAACTGAAAAAA AATACCGTGGATGACATTTCAGAGAGTCTGCGGCAAGGAGGAGG CAAGTTAAACTTTGACGAACTTCGACAAGATCTCAAAGGGAAGG GCCATACTGATGCAGAGATTGAGGCAATATTCACAAAGTACGAC CAAGATGGAGACCAAGAACTGACCGAACATGAACATCAGCAGAT GAGAGACGACTTGGAGAAAGAGAGGGAGGACCT GGATTT GGAT C ACAGTTCTTTACCACGTCCCATGAGCAGCCGAAGTTTCCCTCGAA GC CT GGATGA CTCTG AGGAGGA I GA CGATGA A G AT AGCGGACAT AGCTCCAGAAGGAGGGGAAGCATTTCTAGTGGCGTTTCTTACGAA GAGTTTCAAGTCCTGGTGAGACGAGTGGACCGGATGGAGCATTCC ATCGGCAGCATAGTGTCCAAGATTGACGCCGTGATCGTGAAGCTA GAGATTAT GGAGC GAGCCAAACT GAAGAGGAGGGAGGT GCTGGG AAGGCTGTTGGATGGGGTGGCCGAGGATGAAAGGCTGGGTCGTG ACAGTGAAATCCATAGGGAACAGATGGAACGGCTAGTACGTGAA GAGTTGGAACGCTGGGAATCCGATGATGCAGCTTCCCAGATCAGT CATGGTTTAGGCACGCCAGTGGGACTAAATGGTCAACCTCGCCCC AGAAGCTCCCGCCCATCTTCCTCCCAATCTACAGAAGGCATGGAA GGTGCAGGTGGAAATGGGAGTTCTAATGTCCACGTATGAHuman PKD2: ATGGTCAACTCTAGCAGAGTACAGCCCCAGCAACCTGGGGATGC 3 Full Length; CAAGCGTCCTCCTGCCCCAAGAGCCCCAGACCCTGGAAGATTAAT Codon- GGCCGGCTGCGCCGCCGTGGGAGCCTCCCTCGCCGCTCCTGGGGG Optimized CCTCTGTGAACAGCGGGGCCTGGAAATTGAGATGCAAAGAATCA GACAGGCGGCCGCCAGGGACCCCCCGGCGGGAGCTGCCGCCTCT CCTTCACCTCCACTGTCTAGCTGCAGCAGACAAGCGTGGTCTCGG GATAATCCAGGCTTTGAGGCAGAAGAAGAGGAAGAAGAAGTGGA AGG AG AGGAGGGCGGG A FGGTGGT AGAAATGG AT GT GGA GT GG AGGCCTGGGAGTAGACGGAGTGCCGCTAGTTCCGCTGTGTCCAGC GTGGGAGCCCGGTCTCGGGGCCTAGGGGGCTATCACGGCGCAGG ACACCCTTCCGGAAGAAGAAGACGGCGAGAGGACCAGGGCCCAC CTTGCCCGTCCCCTGTGGGAGGGGGTGACCCACTGCACAGGCACC TGCCCCTCGAAGGCCAGCCACCTAGAGTGGCTTGGGCTGAGAGG CTGGTGCGAGGGCTGAGAGGCCTGTGGGGAACGAGACTGATGGA GGAGAGCAGCACAAACAGAGAGAAGTACCTGAAAAGCGTGCTAA GAGAGCTGGTGACTTACCTTCTTTTCCTGATAGTTCTCTGCATCCT GACATACGGAATGATGAGTTCCAATGTATACTACTACACTCGGAT GATGAGCCAGCTGTTCTTGGACACCCCAGTCTCTAAGACAGAGAA GACCAATTTCAAGACCCTGTCCAGCATGGAGGACTTCTGGAAATT CACAGAAGGCTCGCTGCTCGATGGGTTGTACTGGAAGATGCAGCC TAGCAACCAGACGGAAGCAGATAACAGGTCTTTCATCTTTTATGA GAACTTGCTGCTGGGGGTGCCGAGAATCCGGCAGCTCAGAGTGA GAAATGGGAGCTGTAGCATTCCTCAGGATCTGCGGGATGAGATC AAGGAATGCTATGATGTCTACTCTGTGAGCAGTGAAGACAGGGC CCCCTTTGGCCCTCGGAACGGCACGGCCTGGATTTACACATCTGA GAAGGACTTAAACGGCAGTTCTCACTGGGGCATCATCGCCACCTATAGCGGCGCTGGCTACTACCTAGACCTCTCGCGGACAAGAGAAGAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO AGACCGCCGCCCAGGTCGCCTCTCTGAAGAAAAATGTCTGGCTGG ACCGAGGCACCAGAGCCACTTTCATTGACTTTTCAGTCTACAATG CCAACATAAACCTGTTCTGTGTTGTGAGACTGTTGGTGGAATTCC CAGCTACCGGAGGCGTGATCCCTTCCTGGCAATTTCAACCACTCA AGCTCATTCGCTATGTGACCACTTTTGACTTCTTCCTGGCCGCTTG TGAGATCATCTTCTGCTTTTTCATATTCTACTACGTGGTCGAAGAG ATCCTAGAGATCAGAATCCATAAGCTGCACTACTTTCGGAGCTTT TGGAACTGCCTTGACGTTGTGATTGTGGTACTAAGCGTGGTGGCC ATCGGCATCAACATCTACCGCACTAGTAACGTGGAGGTGCTCCTG CAGTTCCTGGAAGATCAGAACACCTTCCCCAATTTTGAACACCTC GCCTACTGGCAGATCCAGTTCAATAATATCGCTGCAGTGACCGTT TTTTTTGTGTGGATCAAACTCTTCAAGTTTATCAACTTCAACCGAA CCATGTCCCAGCTGAGCACCACAATGAGCCGCTGCGCCAAAGAC CTCTTTGGGTTTGCAATCATGTTCTTCATCATTTTCTTGGCCTATGC CCAGCTGGCATACCTGGTATTTGGAACCCAGGTGGACGACTTTTC CACCTTCCAAGAGTGTATCTTCACACAGTTCCGGATAATCCTCGG CGACATCAACTTCGCTGAGATCGAGGAAGCAAACAGGGTCCTGG GCCCAATTTATTTCACCACATTCGTGTTCTTCATGTTTTTTATCCTG CTGAACATGTTCCTCGCAATTATCAATGACACGTATTCAGAAGTT AAGTCTGATCTGGCTCAGCAGAAGGCTGAAATGGAGCTGAGCGA CCTGATCAGGAAGGGTTACCATAAAGCCCTGGTGAAGCTGAAAC TGAAGAAGAACACAGTGGATGACATCAGTGAATCTCTGCGACAA GGCGGAGGCAAGCTGAACTTTGATGAACTACGTCAGGACCTGAA GGGCAAGGGTCACACAGATGCTGAAATTGAAGCCATCTTTACCA AATATGACCAAGATGGAGACCAGGAACTGACCGAGCATGAGCAC CAGCAGATGAGGGACGATTTGGAGAAGGAGCGGGAAGACCTGGA TCTGGACCACTCTTCTTTACCCCGCCCTATGAGTAGTAGATCGTTT CCTAGAAGCCTGGATGACAGCGAGGAGGATGACGACGAAGATTC CGGTCACAGTAGCAGACGTCGGGGTTCTATCAGCTCTGGCGTTTC ATACGAAGAGTTCCAGGTCCTGGTTAGAAGGGTGGACCGCATGG AACATAGTATCGGCTCCATTGTCAGCAAAATCGACGCCGTCATCGrCAAGCTGGAGATAATGGAGCGGGCGAAGCTCAAACGACGGGAG GTGCTGGGAAGACTCCTGGATGGTGTGGCGGAAGATGAGAGACT GGGCAGAGACTCAGAGATCCACAGAGAGCAGATGGAGCGTTTGG TGAGAGAGGAGCTGGAGAGGTGGGAGTCTGATGATGCCGCCAGC CAGATCAGCCACGGGCTGGGGACCCCCGTTGGCCTGAATGGACA GCCTCGCCCTAGAAGCTCCAGACCCTCCTCTAGCCAAAGTACAGA AGGCATGGAGGGGGCTGGCGGAAACGGCTCCTCCAACGTGCATG TGTGAHuman PKD2: ATGGTGAACTCCAGTAGAGTGCAGCCTCAGCAGCCAGGGGATGC 4 Full Length; CAAGAGGCCTCCAGCACCCAGAGCACCAGACCCAGGCAGGCTGA CpG-Depleted TGGCTGGCTGTGCAGCTGTGGGAGCCAGCCTTGCTGCCCCAGGAG GCCTCTGTGAGCAGAGGGGCCTGGAGATTGAGATGCAGAGAATC AGGCAGGCAGCTGCAAGGGAC CCCC CAGCT GGAGCT GCAGCCT C CCCTTCTCCTCCACTCTCATCATGCAGCAGGCAGGCATGGAGCAG AGATAACCCAGGCTTTGAGGCTGAGGAGGAGGAGGAGGAGGTGG AAGGGGAAGAAGGAGGAATGGTGGTGGAGATGGATGTAGAGTGGAGACCAGGCAGCAGGAGGTCAGCTGCCTCCTCAGCTGTCTCCTCAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO TGTGGGAGCAAGGAGCAGGGGGCTTGGGGGCTACCATGGAGCAG GCCACCCATCTGGGAGGAGGAGAAGGAGAGAGGACCAGGGCCC ACCATGCCCCAGCCCAGTTGGAGGAGGGGACCCACTGCATAGAC ACCTCCCCCTGGAAGGGCAGCCACCCAGGGTGGCCTGGGCAGAG AGGCTGGT TAGAGGGCTGAGGGGI CTCTGGGGAACAAGACTCAT GGAGGAAAGCAGCACTAACAGGGAGAAAT ACC rTAAAAGrGTTI TAAGGGAACTGGTCACATACCTCCTTTTTCTCATAGTCTTGTGCAT CTTGACCTATGGCATGATGAGTTCCAATGTGTACTACTACACCAG GATGATGTCACAGCTCTTCCTAGACACCCCAGTGTCCAAAACTGA GAAAACTAACTTTAAAACTCTGTCTTCCATGGAAGACTTCTGGAA GTTCACAGAAGGCTCCTTATTGGATGGGCTGTACTGGAAGATGCA GCCCAGCAACCAGACTGAAGCTGACAACAGGAGTTTCATCTTCTA TGAGAACCTGCTGTTAGGGGTTCCAAGGATAAGGCAACTCAGGG TCAGAAATGGATCCTGCTCTATCCCCCAGGACTTGAGAGATGAAA TTAAAGAGTGCTATGATGTCTACTCTGTCAGTAGTGAAGATAGGG CTCCCTTTGGGCCCAGGAATGGAACAGCTTGGATCTACACAAGTG AAAAAGACTTGAATGGTAGTAGCCACTGGGGAATCATTGCAACTT ATAGTGGAGCTGGCTATTATCTGGATTTGTCAAGAACAAGAGAGG AAACAGCTGCACAAGTTGCAAGCCTCAAGAAAAATGTCTGGCTG GACAGGGGAACCAGGGCAACTTTTATTGACTTCTCAGTGTACAAT GCCAACATTAACCTGTTCTGTGTGGTCAGGTTATTGGTTGAATTCC CAGCAACAGGTGGTGTGATTCCATCTTGGCAATTTCAGCCTTTAA AGCTGATCAGATATGTCACAACTTTTGATTTCTTCCTGGCAGCCTG TGAGATTATCTTTTGTTTCTTTATCTTTTACTATGTGGTGGAAGAG ATATTGGAAATTAGAATTCACAAACTACACTATTTCAGGAGTTTC TGGAATTGTCTGGATGTTGTGATTGTTGTGCTGTCAGTGGTAGCTA TAGGAATTAACATATACAGAACATCAAATGTGGAGGTGCTACTAC AGTTTCTGGAAGATCAAAATACTTTCCCCAACTTTGAGCATCTGG CATATTGGCAGATACAGTTCAACAATATAGCTGCTGTCACAGTAT TTTTTGTCTGGATTAAGCTCTTCAAATTCATCAATTTTAATAGAAC CATGAGCCAGCTCTCAACAACCATGTCTAGATGTGCCAAAGACCT GTTTGGCTTTGCTATTATGTTCTTCATTATTTTCCTAGCATATGCTC AGTTGGCATACCTTGTCTTTGGCACTCAGGTTGATGACTTCAGTAC TTTCCAAGAGTGTATCTTCACTCAATTCAGAATCATTTTGGGAGAT ATCAACTTTGCAGAGATTGAGGAAGCTAATAGGGTTTTGGGACCA ATTTATTTCACTACATTTGTGTTCTTTATGTTCTTCATTCTTTTGAA TATGTTTTTGGCTATCATCAATGATACTTACTCTGAAGTGAAATCT GACTTGGCACAGCAGAAAGCTGAAATGGAACTCTCAGATCTTATC AGAAAGGGCTACCATAAAGCTTTGGTCAAACTAAAACTGAAAAA AAATACAGTGGATGACATTTCAGAGAGTCTGAGGCAAGGAGGAG GCAAGTTAAACTTTGATGAACTTAGGCAAGATCTCAAAGGGAAG GGCCATACTGATGCAGAGATTGAGGCAATATTCACAAAGTATGA CCAAGArGGAGACCAAGAACTGACAGAACATGAACATCAGCAGA TGAGAGATGACTTGGAGAAAGAGAGGGAGGACCTGGATTTGGAT CACAGTTCTTTACCAAGACCCATGAGCAGCAGGAGTTTCCCTAGG AGCCTGGATGACTCTGAGGAGGATGATGATGAAGATTCTGGACA TAGCTCCAGAAGGAGGGGAAGCATTTCTAGTGGAGTTTCTTATGAAGA GT FTCAAGTCCTGGT GAGAAGGGTGGACAGGATGGAGCAT IAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO CCATTGGCAGCATAGTGTCCAAGATTGATGCTGTGATTGTGAAGC TAGAGATTATGGAGAGGGCCAAACTGAAGAGGAGGGAGGTGCTG GGAAGGCTGTTGGATGGGGTGGCTGAGGATGAAAGGCTGGGTAG AGACAGTGAAATCCATAGGGAACAGATGGAAAGGCTAGTAAGAG AAGAGTTGGAAAGATGGGAATCTGATGATGCAGCTTCCCAGATC AGTCATGGTTTAGGCACTCCAGTGGGACTAAATGGTCAACCTAGA CCCAGAAGCTCCAGACCATCTTCCTCCCAATCTACAGAAGGCATG GAAGGTGCAGGTGGAAATGGGAGTTCTAATGTCCATGTATGAHuman PKD2: ATGGTCAACTCTAGCAGAGTACAGCCCCAGCAACCTGGGGATGC 5 Full Length; CAAGAGGCCTCCTGCCCCAAGAGCCCCAGACCCTGGAAGATTAA Codon- TGGCTGGCTGTGCTGCAGTGGGAGCCTCCCTGGCAGCTCCTGGGG Optimized; GCCTCTGTGAACAGAGAGGCCTGGAAATTGAGATGCAAAGAATC CpG-Depleted AGACAGGCAGCTGCCAGGGACCCCCCAGCAGGAGCTGCTGCCTC TCCTTCACCTCCACTGTCTAGCTGCAGCAGACAAGCATGGTCTAG GGATAATCCAGGCTTTGAGGCAGAAGAAGAGGAAGAAGAAGTGG AAGGAGAGGAGGGAGGGATGGTGGTAGAAATGGATGTGGAGTG GAGGCCTGGGAGTAGAAGAAGTGCTGCTAGTTCTGCTGTGTCCTC TGTGGGAGCCAGGTC FAGAGGCCTAGGGGGC I ATCATGGAGCAG GACACCCTTCTGGAAGAAGAAGAAGGAGGGAGGACCAGGGCCCA CCTTGCCCATCCCCTGTGGGAGGGGGTGACCCACTGCACAGGCAC CTGCCCCTGGAAGGCCAGCCACCTAGAGTGGCTTGGGCTGAGAG GCTTGTCAGGGGGCTGAGAGGCCTGTGGGGAACTAGACTGATGG AGGAGAGCAGCACAAACAGAGAGAAGTACCTGAAATCTGTGCTA AGAGAGCTGGTGACTTACCTTCTTTTCCTGATAGTTCTCTGCATCC TGACATATGGAATGATGAGTTCCAATGTATACTACTACACTAGAA TGATGAGCCAGCTGTTCTTGGACACCCCAGTCTCTAAGACAGAGA AGACCAATTTCAAGACCCTGTCCAGCATGGAGGACTTCTGGAAAT TCACAGAAGGTTCACTGCTGGATGGGTTGTACTGGAAGATGCAGC CTAGCAACCAGACTGAAGCAGATAACAGGTCTTTCATCTTTTATG AGAACTTGCTGCTGGGGGTGCCAAGAATCAGACAGCTCAGAGTC AGAAATGGGAGCTGTAGCATTCCTCAGGATCTGAGAGATGAGAT CAAGGAATGCTATGATGTCTACTCAGTCAGCAGTGAAGACAGGG CCCCCTTTGGCCCAAGAAATGGCACTGCCTGGATTTACACATCTG AGAAGGACTTAAATGGCAGTTCTCACTGGGGCATCATTGCCACCT ATTCTGGAGCTGGCTACTACCTAGACCTCTCAAGAACAAGAGAAG AGACAGCTGCCCAGGTGGCCTCTCTGAAGAAAAATGTCTGGCTGG ACAGGGGCACCAGAGCCACTTTCATTGACTTTTCAGTCTACAATG CCAACATAAACCTGTTCTGTGTTGTCAGACTGTTGGTGGAATTCC CAGCTACAGGAGGAGTGATCCCTTCCTGGCAATTTCAACCACTCA AGCTCATTAGATATGTGACCACTTTTGACTTCTTCCTGGCTGCTTG TGAGATCATCTTCTGCTTTTTCATATTCTACTATGTGGTGGAAGAG ATCCTAGAGATCAGAATCCATAAGCTGCACTACTTTAGAAGCTTT TGGAACTGCCTTGATGTTGTGATTGTGGTACTATCTGTGGTGGCC ATTGGCATCAACATCTACAGAACTAGTAATGTGGAGGTGCTCCTG CAGTTCCTGGAAGATCAGAACACCTTCCCCAATTTTGAACACCTG GCCTACTGGCAGATCCAGTTCAATAATATTGCTGCAGTGACAGTT TTTTTTGTGTGGATCAAACTCTTCAAGTTTATCAACTTCAACAGGACCATGTCCCAGCTGAGCACCACAATGAGCAGATGTGCCAAAGACAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO CTCTTTGGGTTTGCAATCATGTTCTTCATCATTTTCTTGGCCTATGC CCAGCTGGCATACCTGGTATTTGGAACCCAGGTGGATGACTTTTC CACCTTCCAAGAGTGTATCTTCACACAGTTCAGAATAATCCTGGG AGACATCAACTTTGCTGAGATTGAGGAAGCAAACAGGGTCCTGG GCCCAATTTATTTCACCACATTTGTGTTCTTCATGTTTTTTATCCTG CTGAACATGTTCCTGGCAATTATCAATGACACTTATTCAGAAGTT AAGTCTGATCTGGCTCAGCAGAAGGCTGAAATGGAGCTGTCTGAC CTGATCAGGAAGGGTTACCATAAAGCCCTGGTGAAGCTGAAACT GAAGAAGAACACAGTGGATGACATCAGTGAATCTCTGAGGCAAG GAGGAGGCAAGCTGAACTTTGATGAACTAAGGCAGGACCTGAAG GGCAAGGGTCACACAGATGCTGAAATTGAAGCCATCTTTACCAA ATATGACCAAGATGGAGACCAGGAACTGACAGAGCATGAGCACC AGCAGATGAGGGATGATTTGGAGAAGGAGAGAGAAGACCTGGAT CTGGACCACTCTTCTTTACCAAGACCTATGAGTAGTAGATCATTTC CTAGAAGCCTGGATGACTCTGAGGAGGATGATGATGAAGATTCT GGTCACTCTAGCAGAAGGAGAGGTTCTATCAGCTCTGGAGTTTCA TATGAAGAGTTCCAGGTCCTGGTTAGAAGGGTGGACAGGATGGA ACATAGTATTGGCTCCATTGTCAGCAAAATTGATGCTGTCATTGT CAAGCTGGAGATAATGGAGAGAGCAAAGCTCAAAAGGAGAGAG GTGCTGGGAAGACTCCTGGATGGTGTGGCAGAAGATGAGAGACT GGGCAGAGAC FCAGAGA’T CC ACAGAGAGCAGATGGAGAGAC FTG TGAGAGAGGAGCTGGAGAGGTGGGAGTCTGATGATGCTGCCAGC CAGATCAGCCATGGGCTGGGGACCCCAGTTGGCCTGAATGGACA GCCTAGGCCTAGAAGCTCCAGACCCTCCTCTTCCCAATCAACTGA GGGCAT GGAGGGGGCTGGAGGAAAT GGCTCCT CCAAT GT GC AT G TGTGAHuman PKD2: ATGGTGAACTCCAGTCGCGTGCAGCCTCAGCAGCCCGGGGACGC 6 Full Length; CAAGCGGCCGCCCGCGCCCCGCGCGCCGGACCCGGGCCGGCTGA TGGSGG- TGGCTGGCTGCGCGGCCGTGGGCGCCAGCCTCGCCGCCCCGGGCG 3XHA GCCTCTGCGAGCAGCGGGGCCTGGAGATCGAGATGCAGCGCATC CGGCAGGCGGCCGCGCGGGACCCCCCGGCCGGAGCCGCGGCCTC CCCTTCTCCTCCGCTCTCGTCGTGCTCCCGGCAGGCGTGGAGCCG CGATAACCCCGGCTTCGAGGCCGAGGAGGAGGAGGAGGAGGTGG AAGGGGAAGAAGGCGGAAFGGTGGTGGAGATGGACGTAGAGFG GCGCCCGGGCAGCCGGAGGTCGGCCGCCTCCTCGGCCGTGAGCTC CGTGGGCGCGCGGAGCCGGGGGCTTGGGGGCTACCACGGCGCGG GCCACCCGAGCGGGAGGCGGCGCCGGCGAGAGGACCAGGGCCCG CCGTGCCCCAGCCCAGTCGGCGGCGGGGACCCGCTGCATCGCCAC CTCCCCCTGGAAGGGCAGCCGCCCCGAGTGGCCTGGGCGGAGAG GCTGGTTCGCGGGCTGCGAGGTCTCTGGGGAACAAGACTCATGG AGGAAAGCAGCACTAACCGAGAGAAATACCTTAAAAGTGTTTTA CGGGAACTGGTCACATACCTCCTTTTTCTCATAGTCTTGTGCATCT TGACCTACGGCATGATGAGCTCCAATGTGTACTACTACACCCGGA TGATGTCACAGCTCTTCCTAGACACCCCCGTGTCCAAAACGGAGA AAACTAACTTTAAAACTCTGTCTTCCATGGAAGACTTCTGGAAGT TCACAGAAGGCTCCTTATTGGATGGGCTGTACTGGAAGATGCAGC CCAGCAACCAGACTGAAGCTGACAACCGAAGTTTCATCTTCTATGAGAACCTGCTGTTAGGGGTTCCACGAATACGGCAACTCCGAGTCAAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO GAAATGGATCCTGCTCTATCCCCCAGGACTTGAGAGATGAAATTA AAGAGTGCTATGATGTCTACTCTGTCAGTAGTGAAGATAGGGCTC CCTTTGGGCCCCGAAATGGAACCGCTTGGATCTACACAAGTGAAA AAGACTTGAATGGTAGTAGCCACTGGGGAATCATTGCAACTTATA GTGGAGCTGGCTATTATCTGGATTTGTCAAGAACAAGAGAGGAA ACAGCTGCACAAGTTGCTAGCCTCAAGAAAAATGTCTGGCTGGAC CGAGGAACCAGGGCAACTTTTATTGACTTCTCAGTGTACAACGCC AACATTAACCTGTTCTGTGTGGTCAGGTTATTGGTTGAATTCCCAG CAACAGGTGGTGTGATTCCATCTTGGCAATTTCAGCCTTTAAAGC TGATCCGATATGTCACAACTTTTGATTTCTTCCTGGCAGCCTGTGA GATTATCTTTTGTTTCTTTATCTTTTACTATGTGGTGGAAGAGATA TTGGAAATTCGCATTCACAAACTACACTATTTCAGGAGTTTCTGG AATTGTCTGGATGTTGTGATCGTTGTGCTGTCAGTGGTAGCTATA GGAATTAACATATACAGAACATCAAATGTGGAGGTGCTACTACA GTTTCTGGAAGATCAAAATACTTTCCCCAACTTTGAGCATCTGGC ATATTGGCAGATACAGTTCAACAATATAGCTGCTGTCACAGTATT TTTTGTCTGGATTAAGCTCTTCAAATTCATCAATTTTAACAGGACC ATGAGCCAGCTCTCGACAACCATGTCTCGATGTGCCAAAGACCTG TTTGGCTTTGCTATTATGTTCTTCATTATTTTCCTAGCGTATGCTCA GTTGGCATACCTTGTCTTTGGCACTCAGGTCGATGACTTCAGTACT TTCCAAGAGTGTATCTTCACTCAATTCCGTATCATTTTGGGCGATA TCAACTTTGCAGAGATTGAGGAAGCTAATCGAGTTTTGGGACCAA TTTATTTCACTACATTTGTGTTCTTTATGTTCTTCATTCTTTTGAAT ATGTTTTTGGCTATCATCAATGATACTTACTCTGAAGTGAAATCTG ACTTGGCACAGCAGAAAGCTGAAATGGAACTCTCAGATCTTATCA GAAAGGGCTACCATAAAGCTTTGGTCAAACTAAAACTGAAAAAA AATACCGTGGATGACATTTCAGAGAGTCTGCGGCAAGGAGGAGG CAAGTTAAACTTTGACGAACTTCGACAAGATCTCAAAGGGAAGG GCCATACTGATGCAGAGATTGAGGCAATATTCACAAAGTACGAC CAAGATGGAGACCAAGAACTGACCGAACATGAACATCAGCAGAT GAGAGACGACTTGGAGAAAGAGAGGGAGGACCT GGATTT GGAT C ACAGTTCTTTACCACGTCCCATGAGCAGCCGAAGTTTCCCTCGAA GCCTGGATGACTCTGAGGAGGATGACGATGAAGATAGCGGACAT AGCTCCAGAAGGAGGGGAAGCATTTCTAGTGGCGTTTCTTACGAA GAGTTTCAAGTCCTGGTGAGACGAGTGGACCGGATGGAGCATTCC ATCGGCAGCATAGTGTCCAAGATTGACGCCGTGATCGTGAAGCTA GAGA FT AT GGAGC GAGCCA A A CTGA A G AGG AGGG AGG FGC I GGG AAGGCTGTTGGATGGGGTGGCCGAGGATGAAAGGCTGGGTCGTG ACAGTGAAATCCATAGGGAACAGATGGAACGGCTAGTACGTGAA GAGTTGGAACGCTGGGAATCCGATGATGCAGCTTCCCAGATCAGT CATGGTTTAGGCACGCCAGTGGGACTAAATGGTCAACCTCGCCCC AGAAGCTCCCGCCCATCTTCCTCCCAATCTACAGAAGGCATGGAA GGTGCAGGTGGAAATGGGAGTTCTAATGTCCACGTAACCGGCGG CAGCGGCGGCTACCCATACGATGTTCCAGATTACGCTTACCCATA CGATGTTCCAGATTACGCTTACCCATACGATGTTCCAGATTACGC TTGAHuman PKD2: ATGGTCAACTCTAGCAGAGTACAGCCCCAGCAACCTGGGGATGC 7Full Length; CAAGCGTCCTCCTGCCCCAAGAGCCCCAGACCCTGGAAGATTAATAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NOCodon- GGCCGGCTGCGCCGCCGTGGGAGCCTCCCTCGCCGCTCCTGGGGG Optimized; CCTCTGTGAACAGCGGGGCCTGGAAATTGAGATGCAAAGAATCA TGGSGG- GACAGGCGGCCGCCAGGGACCCCCCGGCGGGAGCTGCCGCCTCT 3XHA CCTTCACCTCCACTGTCTAGCTGCAGCAGACAAGCGTGGTCTCGG GAFAATCCAGGC FTTGAGGCAGAAGAAGAGGAAGAAGAAGTGGA AGGAGA GGA GGGCGGGATGG FGGTAGAAATGGATGTGGAGTGG AGGCCTGGGAGTAGACGGAGTGCCGCTAGTTCCGCTGTGTCCAGC GTGGGAGCCCGGTCTCGGGGCCTAGGGGGCTATCACGGCGCAGG ACACCCTTCCGGAAGAAGAAGACGGCGAGAGGACCAGGGCCCAC CTTGCCCGTCCCCTGTGGGAGGGGGTGACCCACTGCACAGGCACC TGCCCCTCGAAGGCCAGCCACCTAGAGTGGCTTGGGCTGAGAGG CTGGTGCGAGGGCTGAGAGGCCTGTGGGGAACGAGACTGATGGA GGAGAGCAGCACAAACAGAGAGAAGTACCTGAAAAGCGTGCTAA GAGAGCTGGTGACTTACCTTCTTTTCCTGATAGTTCTCTGCATCCT GACATACGGAATGATGAGTTCCAATGTATACTACTACACTCGGAT GATGAGCCAGCTGTTCTTGGACACCCCAGTCTCTAAGACAGAGAA GACCAATTTCAAGACCCTGTCCAGCATGGAGGACTTCTGGAAATT CACAGAAGGCTCGCTGCTCGATGGGTTGTACTGGAAGATGCAGCC TAGCAACCAGACGGAAGCAGATAACAGGTCTTTCATCTTTTATGA GAACTTGCTGCTGGGGGTGCCGAGAATCCGGCAGCTCAGAGTGA GA A AT GGG AGCT GIA GC ATTCCT CA GGA T CT GCGGG A I GA GAT C AAGGAATGCTATGATGTCTACTCTGTGAGCAGTGAAGACAGGGC CCCCTTTGGCCCTCGGAACGGCACGGCCTGGATTTACACATCTGA GAAGGACTTAAACGGCAGTTCTCACTGGGGCATCATCGCCACCTA TAGCGGCGCTGGCTACTACCTAGACCTCTCGCGGACAAGAGAAG AGACCGCCGCCCAGGTCGCCTCTCTGAAGAAAAATGTCTGGCTGG ACCGAGGCACCAGAGCCACTTTCATTGACTTTTCAGTCTACAATG CCAACATAAACCTGTTCTGTGTTGTGAGACTGTTGGTGGAATTCC CAGCTACCGGAGGCGTGATCCCTTCCTGGCAATTTCAACCACTCA AGCTCATTCGCTATGTGACCACTTTTGACTTCTTCCTGGCCGCTTG TGAGATCATCTTCTGCTTTTTCATATTCTACTACGTGGTCGAAGAG ATCCTAGAGATCAGAATCCATAAGCTGCACTACTTTCGGAGCTTT TGGAACTGCCTTGACGTTGTGATTGTGGTACTAAGCGTGGTGGCC ATCGGCATCAACATCTACCGCACTAGTAACGTGGAGGTGCTCCTG CAGTTCCTGGAAGATCAGAACACCTTCCCCAATTTTGAACACCTC GCCTACTGGCAGATCCAGTTCAATAATATCGCTGCAGTGACCGTT TTTTTTGTGTGGATCAAACTCTTCAAGTTTATCAACTTCAACCGAA CCATGTCCCAGCTGAGCACCACAATGAGCCGCTGCGCCAAAGAC CTCTTTGGGTTTGCAATCATGTTCTTCATCATTTTCTTGGCCTATGC CCAGCTGGCATACCTGGTATTTGGAACCCAGGTGGACGACTTTTC CACCTTCCAAGAGTGTATCTTCACACAGTTCCGGATAATCCTCGG CGACATCA A CTTCGCTG AG A I CGAGGAAGC A A ACAGGGTCCT GG GCCCAATTTATTTCACCACATTCGTGTTCTTCATGTTTTTTATCCTG CTGAACATGTTCCTCGCAATTATCAATGACACGTATTCAGAAGTT AAGTCTGATCTGGCTCAGCAGAAGGCTGAAATGGAGCTGAGCGA CCTGATCAGGAAGGGTTACCATAAAGCCCTGGTGAAGCTGAAAC TGAAGAAGAACACAGTGGATGACATCAGTGAATCTCTGCGACAAGGCGGAGGCAAGCTGAAC FTTGAT GAACTACGTCAGGACC I GAAAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO GGGCAAGGGTCACACAGATGCTGAAATTGAAGCCATCTTTACCA AATATGACCAAGATGGAGACCAGGAACTGACCGAGCATGAGCAC CAGCAGATGAGGGACGATTTGGAGAAGGAGCGGGAAGACCTGGA TCTGGACCACTCTTCTTTACCCCGCCCTATGAGTAGTAGATCGTTT CCTAGAAGCCTGGAT GACAGCGAGGAGGAT GACG ACGAAGAT’I C CGGTCACAGTAGCAGACGTCGGGGTTCTATCAGCTCTGGCGTTTC ATACGAAGAGTTCCAGGTCCTGGTTAGAAGGGTGGACCGCATGG AACATAGTATCGGCTCCATTGTCAGCAAAATCGACGCCGTCATCG TCAAGCTGGAGATAATGGAGCGGGCGAAGCTCAAACGACGGGAG GFGC I GGGAAGACTCC’I GGATGGT GTGGCGGAAGATGAGAGAC F GGGCAGAGAC FCAGAGA’T CCACAGAGAGCAGATGGAGCGT’T TGG TGAGAGAGGAGCTGGAGAGGTGGGAGTCTGATGATGCCGCCAGC CAGATCAGCCACGGGCTGGGGACCCCCGTTGGCCTGAATGGACA GCCTCGCCCTAGAAGCTCCAGACCCTCCTCTAGCCAAAGTACAGA AGGCAT GGAGGGGGCTGGCGGAAACGGCTCCTCCAAC GT GC AT G TGACCGGCGGCAGCGGCGGCTACCCATACGATGTTCCAGATTACG CTTACCCATACGATGTTCCAGATTACGCTTACCCATACGATGTTCC AGATTACGCTTGAHuman PKD2: ATGGTGAACTCCAGTAGAGTGCAGCCTCAGCAGCCAGGGGATGC 8 Full Length; CAAGAGGCCTCCAGCACCCAGAGCACCAGACCCAGGCAGGCTGA CpG -Depleted; TGGCTGGCTGTGCAGCTGTGGGAGCCAGCCTTGCTGCCCCAGGAG TGGSGG- GCCT CT GT GAGCAGAGGGGCCT GGAGATT GA GAT GC AGAGAAT C 3XHA AGGCAGGCAGCTGCAAGGGACCCCCCAGCTGGAGCTGCAGCCTC CCCTTCTCCTCCACTCTCATCATGCAGCAGGCAGGCATGGAGCAG AGATAACCCAGGCTTTGAGGCTGAGGAGGAGGAGGAGGAGGTGG AAGGGGAAGAAGGAGGAATGGTGGTGGAGATGGATGTAGAGTG GAGACCAGGCAGCAGGAGGTCAGCTGCCTCCTCAGCTGTCTCCTC TGTGGGAGC AAGGAGCAGGGGGCTTGGGGGCTACC ATGGAGCAG GCCACCCATCTGGGAGGAGGAGAAGGAGAGAGGACCAGGGCCC ACCATGCCCCAGCCCAGTTGGAGGAGGGGACCCACTGCATAGAC ACCTCCCCCTGGAAGGGCAGCCACCCAGGGTGGCCTGGGCAGAG AGGCTGGTTAGAGGGCTGAGGGGTCTCTGGGGAACAAGACTCAT GGAGGAAAGCAGCACTAACAGGGAGAAATACCTTAAAAGTGTTT TAAGGGAACTGGTCACATACCTCCTTTTTCTCATAGTCTTGTGCAT CTTGACCTATGGCATGATGAGTTCCAATGTGTACTACTACACCAG GATGATGTCACAGCTCTTCCTAGACACCCCAGTGTCCAAAACTGA GAAAACTAACTTTAAAACTCTGTCTTCCATGGAAGACTTCTGGAA GTTCACAGAAGGCTCCTTATTGGATGGGCTGTACTGGAAGATGCA GCCCAGCAACCAGACTGAAGCTGACAACAGGAGTTTCATCTTCTA TGAGAACCTGCTGTTAGGGGTTCCAAGGATAAGGCAACTCAGGG TCAGAAATGGATCCTGCTCTATCCCCCAGGACTTGAGAGATGAAA TTAAAGAGTGCTATGATGTCTACTCTGTCAGTAGTGAAGATAGGG CTCCCTTTGGGCCCAGGAATGGAACAGCTTGGATCTACACAAGTG AAAAAGACTTGAATGGTAGTAGCCACTGGGGAATCATTGCAACTT ATAGTGGAGCTGGCTATTATCTGGATTTGTCAAGAACAAGAGAGG AAACAGCTGCACAAGTTGCAAGCCTCAAGAAAAATGTCTGGCTG GACAGGGGAACCAGGGCAACTTTTATTGACTTCTCAGTGTACAATGCCAACATTAACCTGTTCTGTGTGGTCAGGTTATTGGTTGAATTCCAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO CAGCAACAGGTGGTGTGATTCCATCTTGGCAATTTCAGCCTTTAA AGCTGATCAGATATGTCACAACTTTTGATTTCTTCCTGGCAGCCTG TGAGATTATCTTTTGTTTCTTTATCTTTTACTATGTGGTGGAAGAG ATATTGGAAATTAGAATTCACAAACTACACTATTTCAGGAGTTTC TGGAATTGTCTGGATGTTGTGATTGTTGTGCTGTCAGTGGTAGCTA TAGGAATTAACATATACAGAACATCAAATGTGGAGGTGCTACTAC AGTTTCTGGAAGATCAAAATACTTTCCCCAACTTTGAGCATCTGG CATATTGGCAGATACAGTTCAACAATATAGCTGCTGTCACAGTAT TTTTTGTCTGGATTAAGCTCTTCAAATTCATCAATTTTAATAGAAC CATGAGCCAGCTCTCAACAACCATGTCTAGATGTGCCAAAGACCT GTTTGGCTTTGCTATTATGTTCTTCATTATTTTCCTAGCATATGCTC AGTTGGCATACCTTGTCTTTGGCACTCAGGTTGATGACTTCAGTAC TTTCCAAGAGTGTATCTTCACTCAATTCAGAATCATTTTGGGAGAT ATCAACTTTGCAGAGATTGAGGAAGCTAATAGGGTTTTGGGACCA ATTTATTTCACTACATTTGTGTTCTTTATGTTCTTCATTCTTTTGAA TATGTTTTTGGCTATCATCAATGATACTTACTCTGAAGTGAAATCT GACTTGGCACAGCAGAAAGCTGAAATGGAACTCTCAGATCTTATC AGAAAGGGCTACCATAAAGCTTTGGTCAAACTAAAACTGAAAAA AAATACAGTGGATGACATTTCAGAGAGTCTGAGGCAAGGAGGAG GCAAGTTAAACTTTGATGAACTTAGGCAAGATCTCAAAGGGAAG GGCCATACTGATGCAGAGATTGAGGCAATATTCACAAAGTATGA CCAAGATGGAGACCAAGAACTGACAGAACATGAACATCAGCAGA TGAGAGATGACTTGGAGAAAGAGAGGGAGGACCTGGATTTGGAT CACAGTTCTTTACCAAGACCCATGAGCAGCAGGAGTTTCCCTAGG AGCCTGGATGACTCTGAGGAGGATGATGATGAAGATTCTGGACA TAGCTCCAGAAGGAGGGGAAGCATTTCTAGTGGAGTTTCTTATGA AG AG ITT C A AG I C CT GG’I GA GAAGGGT GGAC AGG A I GGA GC Al T CCATTGGCAGCATAGTGTCCAAGATTGATGCTGTGATTGTGAAGC TAGAGATTATGGAGAGGGCCAAACTGAAGAGGAGGGAGGTGCTG GGAAGGCTGTTGGATGGGGTGGCTGAGGATGAAAGGCTGGGTAG AGACAGTGAAATCCATAGGGAACAGATGGAAAGGCTAGTAAGAG AAGAGTTGGAAAGATGGGAATCTGATGATGCAGCTTCCCAGATC AGTCATGGTTTAGGCACTCCAGTGGGACTAAATGGTCAACCTAGA CCCAGAAGCTCCAGACCATCTTCCTCCCAATCTACAGAAGGCATG GAAGGTGCAGGTGGAAATGGGAGTTCTAATGTCCATGTAACCGG CGGCAGCGGCGGCTACCCATACGATGTTCCAGATTACGCTTACCC ATACGATGTTCCAGATTACGCTTACCCATACGATGTTCCAGATTA CGCTTGAHuman PKD2: ATGGTCAACTCTAGCAGAGTACAGCCCCAGCAACCTGGGGATGC 9 Full Length; CAAGAGGCCTCCTGCCCCAAGAGCCCCAGACCCTGGAAGATTAA Codon- TGGCTGGCTGTGCTGCAGTGGGAGCCTCCCTGGCAGCTCCTGGGG Optimized; GCCTCTGTGAACAGAGAGGCCTGGAAATTGAGATGCAAAGAATC CpG -Depleted; AGACAGGCAGCTGCCAGGGACCCCCCAGCAGGAGCTGCTGCCTC TGGSGG- TCCTTCACCTCCACTGTCTAGCTGCAGCAGACAAGCATGGTCTAG 3XHA GGAT A A ICCAGGCT ITGAGGCAGAAGA AGAGGAAGAAGAAG IGG AAGGAGAGGAGGGAGGGATGGTGGTAGAAATGGATGTGGAGTG GAGGCCTGGGAGTAGAAGAAGTGCTGCTAGTTCTGCTGTGTCCTCTGTGGGAGCCAGGTCTAGAGGCCTAGGGGGCTATCATGGAGCAGAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO GACACCCTTCTGGAAGAAGAAGAAGGAGGGAGGACCAGGGCCCA CCTTGCCCATCCCCTGTGGGAGGGGGTGACCCACTGCACAGGCAC CTGCCCCTGGAAGGCCAGCCACCTAGAGTGGCTTGGGCTGAGAG GCTT GT C AGGGGGCTGAGAGGC CT GT GGGGAACTAGACT GAT GG AGGAGAGCAGCACAAACAGAGAGAAG’T ACC I GAAATCTGT GC’I A AGAGAGCTGGTGACTTACCTTCTTTTCCTGATAGTTCTCTGCATCC TGACATATGGAATGATGAGTTCCAATGTATACTACTACACTAGAA TGATGAGCCAGCTGTTCTTGGACACCCCAGTCTCTAAGACAGAGA AGACCAATTTCAAGACCCTGTCCAGCATGGAGGACTTCTGGAAAT TCACAGAAGGTTCACTGCTGGATGGGTTGTACTGGAAGATGCAGC CTAGCAACCAGACTGAAGCAGATAACAGGTCTTTCATCTTTTATG AGAACTTGCTGCTGGGGGTGCCAAGAATCAGACAGCTCAGAGTC AGAAATGGGAGCTGTAGCATTCCTCAGGATCTGAGAGATGAGAT CAAGGAATGCTATGATGTCTACTCAGTCAGCAGTGAAGACAGGG CCCCCTTTGGCCCAAGAAATGGCACTGCCTGGATTTACACATCTG AGAAGGACTTAAATGGCAGTTCTCACTGGGGCATCATTGCCACCT ATTCTGGAGCTGGCTACTACCTAGACCTCTCAAGAACAAGAGAAG AGACAGCTGCCCAGGTGGCCTCTCTGAAGAAAAATGTCTGGCTGG ACAGGGGCACCAGAGCCACTTTCATTGACTTTTCAGTCTACAATG CCAACATAAACCTGTTCTGTGTTGTCAGACTGTTGGTGGAATTCC CAGCTACAGGAGGAGTGATCCCTTCCTGGCAATTTCAACCACTCA AGCTCATTAGATATGTGACCACTTTTGACTTCTTCCTGGCTGCTTG TGAGATCATCTTCTGCTTTTTCATATTCTACTATGTGGTGGAAGAG ATCCTAGAGATCAGAATCCATAAGCTGCACTACTTTAGAAGCTTT TGGAACTGCCTTGATGTTGTGATTGTGGTACTATCTGTGGTGGCC ATTGGCATCAACATCTACAGAACTAGTAATGTGGAGGTGCTCCTG CAGTTCCTGGAAGATCAGAACACCTTCCCCAATTTTGAACACCTG GCCTACTGGCAGATCCAGTTCAATAATATTGCTGCAGTGACAGTT TTTTTTGTGTGGATCAAACTCTTCAAGTTTATCAACTTCAACAGGA CCATGTCCCAGCTGAGCACCACAATGAGCAGATGTGCCAAAGAC CTCTTTGGGTTTGCAATCATGTTCTTCATCATTTTCTTGGCCTATGC CCAGCTGGCATACCTGGTATTTGGAACCCAGGTGGATGACTTTTC CACCTTCCAAGAGTGTATCTTCACACAGTTCAGAATAATCCTGGG AGACATCAACTTTGCTGAGATTGAGGAAGCAAACAGGGTCCTGG GCCCAATTTATTTCACCACATTTGTGTTCTTCATGTTTTTTATCCTG CTGAACATGTTCCTGGCAATTATCAATGACACTTATTCAGAAGTT AAGFC I GATCTGGCTCAGCAGA AGGCTGAAAFGGAGC’I GTCTGAC CTGATCAGGAAGGGTTACCATAAAGCCCTGGTGAAGCTGAAACT GAAGAAGAACACAGTGGATGACATCAGTGAATCTCTGAGGCAAG GAGGAGGCAAGCTGAACTTTGATGAACTAAGGCAGGACCTGAAG GGCAAGGGTCACACAGATGCTGAAATTGAAGCCATCTTTACCAA ATATGACCAAGATGGAGACCAGGAACTGACAGAGCATGAGCACC AGCAGATGAGGGATGATTTGGAGAAGGAGAGAGAAGACCTGGAT CTGGACCACTCTTCTTTACCAAGACCTATGAGTAGTAGATCATTTC CTAGAAGCCTGGATGACTCTGAGGAGGATGATGATGAAGATTCT GGTCACTCTAGCAGAAGGAGAGGTTCTATCAGCTCTGGAGTTTCA TATGAAGAGTTCCAGGTCCTGGTTAGAAGGGTGGACAGGATGGAACATAGTATTGGCTCCATTGTCAGCAAAATTGATGCTGTCATTGTAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO CAAGCTGGAGATAATGGAGAGAGCAAAGCTCAAAAGGAGAGAG GTGCTGGGAAGACTCCTGGATGGTGTGGCAGAAGATGAGAGACT GGGCAGAGACTCAGAGATCCACAGAGAGCAGATGGAGAGACTTG TGAGAGAGGAGCTGGAGAGGTGGGAGTCTGATGATGCTGCCAGC CAGATCAGCCATGGGCTGGGGACCCCAGTTGGCCTGAATGGACA GCCTAGGCCTAGAAGCTCCAGACCCTCCTCTTCCCAATCAACTGA GGGCATGGAGGGGGCTGGAGGAAATGGCTCCTCCAATGTGCATG TGACCGGCGGCAGCGGCGGCTACCCATACGATGTTCCAGATTACG CTTACCCATACGATGTTCCAGATTACGCTTACCCATACGATGTTCC AGATTACGCTTGAMouse PKD2 ATGGTTAACTCCAGACGCGTGCAGCCGCAGCCGCCCGGGGACGC 10 GGGACGCTCGCCCGCGCCGCGAGCGTCCGGACCCGGGCGCCTGG TGGCGGGAGGCGCCGGCCTAGCTGTCCCCGGCGGCCTCGGGGAG CAGCGGGGCCTGGAGATCGAGATGGAGCGCATCCGGCAGGCGGC CGCTCGGGATCCCCCGGCCGGAGCCTCGGCCTCGCCGTCTCCTCC GCTTTCGTCCTGCTCCAGGCAAGCGTGGAGCCGCGACAACCCGGG CTTT GAGGCAGAGGAGGAT GACGACGACGACGAGGT GGAAGGAG AAGAAGGAGGGATGGTGGTAGAGATGGATGTGGAGTGGCGCCCG GGCAGTCGGAGGFCGGCCTCCTCC’T CGGCCGTGAGC FCGGT GGGC GCCCGCGGCCGAGGGCTCGGGAGCTACCGCGGCGCGGCTCACCT GAGCGGGAGGCGGCGCCGGCTAGAGGACCAGGGCGCGCAGTGTC CCAGCCCCGCGGGCGGCGGGGACCCGCTGCATCGCCACCTCCCGC TGGAGGGCCAGCC ACCC CGAGT GGC CTGGGC AGAGAGGCT GGTG CGAGGGCTGCGAGGTCTCTGGGGAACAAGACTCATGGAAGAGAG CAACGCCAACCGAGAGAAGTACCTGAAAAGTGTGTTACGGGAGC TGGTCACTTACCTCTTTTTCCTCGTAGTCTTGTGCATCTTGACCTA CGGCATGATGAGCTCCAATGTGTACTACTACACTCGGACACTGTC ACAGCTATTCATAGACACCCCAGTGTCGAAAACAGAGAAAACCA ACTTTAAAACTCTTTCTTCCATGGAGGACTTCTGGAAGTTCACCG AAGGCTCCTTCCTGGATGGGCTGTACTGGAAGGCACAGACCAGC AACCACACGCAAGCTGACAACCGAAGCTTTATCTTCTATGAGAAC CTGCTGCTAGGAGTGCCGCGTCTACGCCAACTCCGAGTCAGAAAC GGATCCTGCTCCATCCCTCAGGACCTGCGAGATGAAATTAAAGAG TGCTATGACGTCTACTCCGTCAGCAGTGAGGACAGAGCTCCATTT GGACCGCGGA AT GGA ACT GC G I GGATG FAC AC A AGT GAGAAGGA GCTGAATGGGAGCAGTCACTGGGGGATCATTGCGTCGTACAGTG GAGCGGGTTACTACCTGGATCTGTCCAGAACCAGGGAGGAGACA GCAGCCCAGCTTGCTGGCCTCAGGAGGAACTTCTGGCTGGACCGG GGCACGCGGGCAGCTTTTATAGACTTCTCGGTGTATAACGCAAAC ATTAACCTGTTCTGTGTGGTCAGGTTATTGGCGGAGTTCCCAGCA ACGGGTGGCGTGGTACCCTCTTGGCAGTTTCAGCCTGTAAAACTG ATCCGCTATGTCACAGCCTTTGATTTCTTCCTGGCAGCCTGTGAGA TCATCTTTTGTTTCTTTATCATTTACTATGTGGTGGAAGAGATATT GGAAATTCGGATTCACAGACTGAGCTATTTCAGGAGTTTCTGGAA TTGrCTGGATGrTGTGATFGTCGTGFTATCTGTAGTAGCTATGGTG ATTAACATTTACCGAATGTCAAATGCAGAGGGGCTGCTACAGTTT CTTGAAGATCAAAATTCTTTCCCCAACTTTGAGCATGTGGCATACTGGCAAATACAGTTCAACAATATAAGTGCTGTCATGGTATTTTTGAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO GTCTGGATTAAGCTCTTCAAATTCATCAATTTTAATAGGACCATG AGCCAGCTCTCCACAACCATGTCTCGATGTGCCAAAGACCTCTTC GGCTTCACCATAATGTTCTCCATCATCTTCTTGGCATACGCACAGC TGGCATACCTTGTCTTCGGCACCCAGGTCGATGACTTCAGCACTTT CCAAGAATGTATCTTCACCCAGTTCCGCATCATTTTGGGTGATATC AACTTCGCAGAGATCGAGGAAGCTAACCGAGTTTTGGGGCCACTT TATTTTACTACATTTGTGTTCTTTATGTTCTTCATTCTTTTGAATAT GTTCCTGGCGATCATCAATGATTCGTACTCTGAAGTGAAATCCGA TCTGGCCCAGCAGAAAGCAGAAATGGAACTCTCAGACCTTATCA GAAAGGGCTGCCAAAAAGCAC’I GGTCAAACTAAAACTGAAAAGA AACAC rGTAGATGCCATCTCAGAGAGrC I CCGGCAAGGTGGI GGC AAACTGAACTTTGATGAGCTTCGGCAAGACCTGAAAGGGAAGGG CCATACAGATGCAGAGATTGAGGCCATATTCACTAAATATGACCA GGATGGCGACCAGGAACTGACCGAGCGTGAGCATCAACAGATGA GAGATGACTTGGAGAAAGAGAGGGAGGACCTAGACTTGGAACAC AGCTCTTTACCACGTCCGATGAGCAGCAGAAGTTTCCCCAGAAGC CTGGATGACTCCGAGGAGGAGGATGACGAAGACAGTGGCCATAG CTCCAGGAGGAGGGGAAGCATCTCCAGTGGGGTTTCCTATGAAG AGTTCCAAGTACTGGTGAGGCGCGTGGACCGCATGGAGCACTCC ATTGGCAGCATCGTTTCCAAGATTGATGCCGTGATTGTCAAGCTG GA GAT CA T GG AGCGGGCC A AGC I GA A GA GA CGA GA GGTG FT A GG ACGGCTGCTGGATGGCGTGGCTGAGGATGCGCGACTGGGTCGGG ACAGTGAGATCCACAGGGAGCAGATGGAGCGCCTGGTGCGGGAA GAGCTGGAGCGCTGGGAATCGGATGATGCAGCTTCGCAAACAGG TCATGGTGTAAGCACACAAGTGGGACTCGGTGGCCAGCCCCACCC CAGAAACCCGCGCCCTCCTTCCTCCCAGTCTGCAGAGGGCCTGGA AGGTGGAGGTGGAAATGGAAGTGCCAACGTCCATGCCTAAMouse PKD2- ATGGTTAACTCCAGACGCGTGCAGCCGCAGCCGCCCGGGGACGC 11 TGGSGG- GGGACGCTCGCCCGCGCCGCGAGCGTCCGGACCCGGGCGCCTGG 3XHA TGGCGGGAGGCGCCGGCCTAGCTGTCCCCGGCGGCCTCGGGGAG CAGCGGGGCCTGGAGATCGAGATGGAGCGCATCCGGCAGGCGGC CGCTCGGGATCCCCCGGCCGGAGCCTCGGCCTCGCCGTCTCCTCC GCTTTCGTCCTGCTCCAGGCAAGCGTGGAGCCGCGACAACCCGGG CT I TG AGGCA GA GGA GG AT GACG ACGA CGA CGAGGTGG A AGG AG AAGAAGGAGGGATGGTGGTAGAGATGGATGTGGAGTGGCGCCCG GGCAGTCGGAGGTCGGCCTCCTCCTCGGCCGTGAGCTCGGTGGGC GCCCGCGGCCGAGGGCTCGGGAGCTACCGCGGCGCGGCTCACCT GAGCGGGAGGCGGCGCCGGCTAGAGGACCAGGGCGCGCAGTGTC CCAGCCCCGCGGGCGGCGGGGACCCGCTGCATCGCCACCTCCCGC TGGAGGGCCAGCCACCCCGAGTGGCCTGGGCAGAGAGGCTGGI G CGAGGGCTGCGAGGTCTCTGGGGAACAAGACTCATGGAAGAGAG CAACGCCAACCGAGAGAAGTACCTGAAAAGTGTGTTACGGGAGC TGGTCACTTACCTCTTTTTCCTCGTAGTCTTGTGCATCTTGACCTA CGGCATGATGAGCTCCAATGTGTACTACTACACTCGGACACTGTC ACAGCTATTCATAGACACCCCAGTGTCGAAAACAGAGAAAACCA ACTTTAAAACTCTTTCTTCCATGGAGGACTTCTGGAAGTTCACCG AAGGCTCCTTCCTGGATGGGCTGTACTGGAAGGCACAGACCAGCAACCACACGCAAGCTGACAACCGAAGCTTTATCTTCTATGAGAACAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO CTGCTGCTAGGAGTGCCGCGTCTACGCCAACTCCGAGTCAGAAAC GGATCCTGCTCCATCCCTCAGGACCTGCGAGATGAAATTAAAGAG TGCTATGACGTCTACTCCGTCAGCAGTGAGGACAGAGCTCCATTT GGAC C GCGGAAT GGA ACT GC GT GGAT GT AC AC AA GT GAGAAGGA GCTGAATGGGAGCAGTCACTGGGGGATCATTGCGTCGTACAGTG GAGCGGGT TACT ACCT GGAT Cl GTCCAGAACCAGGGAGGAGACA GCAGCCCAGCTTGCTGGCCTCAGGAGGAACTTCTGGCTGGACCGG GGCACGCGGGCAGCTTTTATAGACTTCTCGGTGTATAACGCAAAC ATTAACCTGTTCTGTGTGGTCAGGTTATTGGCGGAGTTCCCAGCA ACGGGTGGCGTGGTACCCTCTTGGCAGTTTCAGCCTGTAAAACTG ATCCGCTATGTCACAGCCTTTGATTTCTTCCTGGCAGCCTGTGAGA TCATCTTTTGTTTCTTTATCATTTACTATGTGGTGGAAGAGATATT GGAAATTCGGATTCACAGACTGAGCTATTTCAGGAGTTTCTGGAA TTGTCTGGATGTTGTGATTGTCGTGTTATCTGTAGTAGCTATGGTG ATTAACATTTACCGAATGTCAAATGCAGAGGGGCTGCTACAGTTT CTTGAAGATCAAAATTCTTTCCCCAACTTTGAGCATGTGGCATAC TGGCAAATACAGTTCAACAATATAAGTGCTGTCATGGTATTTTTG GTCTGGATTAAGCTCTTCAAATTCATCAATTTTAATAGGACCATG AGCCAGCTCTCCACAACCATGTCTCGATGTGCCAAAGACCTCTTC GGCTTCACCATAATGTTCTCCATCATCTTCTTGGCATACGCACAGC TGGCATACCTTGTCTTCGGCACCCAGGTCGATGACTTCAGCACTTT CCAAGAATGTATCTTCACCCAGTTCCGCATCATTTTGGGTGATATC AACTTCGCAGAGATCGAGGAAGCTAACCGAGTTTTGGGGCCACTT TATTTTACTACATTTGTGTTCTTTATGTTCTTCATTCTTTTGAATAT GTTCCTGGCGATCATCAATGATTCGTACTCTGAAGTGAAATCCGA TCTGGCCCAGCAGAAAGCAGAAATGGAACTCTCAGACCTTATCA GAAAGGGCTGCCAAAAAGCACl GGTCAAACTAAAACTGAAAAGA AACACTGTAGATGCCATCTCAGAGAGTCTCCGGCAAGGTGGTGGC AAACTGAACTTTGATGAGCTTCGGCAAGACCTGAAAGGGAAGGG CCATACAGATGCAGAGATTGAGGCCATATTCACTAAATATGACCA GGAT GGCGACC AGGAACTGACCGAGCGT GAGCAT CAACAGAT GA GAGA I'GAC’T TGGAGAAAGAGAGGGAGGACCT AGAC I'TGGAACAC AGCTCTTTACCACGTCCGATGAGCAGCAGAAGTTTCCCCAGAAGC CTGGATGACTCCGAGGAGGAGGATGACGAAGACAGTGGCCATAG CTCCAGGAGGAGGGGAAGCATCTCCAGTGGGGTTTCCTATGAAG AGTTCCAAGTACTGGTGAGGCGCGTGGACCGCATGGAGCACTCC ATTGGCAGCATCGTTTCCAAGATTGATGCCGTGATTGTCAAGCTG GAGATCATGGAGCGGGCCAAGCTGAAGAGACGAGAGGTGTTAGG ACGGCTGCTGGATGGCGTGGCTGAGGATGCGCGACTGGGTCGGG ACAGTGAGATCCACAGGGAGCAGATGGAGCGCCTGGTGCGGGAA GAGCTGGAGCGCTGGGAATCGGATGATGCAGCTTCGCAAACAGG TCATGGTGTAAGCACACAAGTGGGACTCGGTGGCCAGCCCCACCC CAGAAACCCGCGCCCTCCTTCCTCCCAGTCTGCAGAGGGCCTGGA AGGTGGAGGTGGAAATGGAAGTGCCAACGTCCATGCCACCGGCG GCAGCGGCGGCTACCCATACGATGTTCCAGATTACGCTTACCCAT ACGATGTTCCAGATTACGCTTACCCATACGATGTTCCAGATTACGCTTAAAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO CBA Promoter CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAA 12 CGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGT AACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTT ACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCC AAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTG GCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCA GTACATCTACTCGAGGCCACGTTCTGCTTCACTCTCCCCATCTCCC CCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTTAATTATTT T GT GC AGCGAT GGGGGC GGGGGGGGGGGGGGGGC GCGCGC C AG GCGGGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAG AGGTGCGGCCTCTCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTC CTTTTATGGCGAGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGA AGCGCGCGGCGGGCGGGAGCGGGATCAGCCACCGCGGTGGCGGC CTAGAGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGTAA GTTTAGTCTTTTTGTCTTTTATTTCAGGTCCCGGATCCGGTGGTGG TGCAAATCAAAGAACTGCTCCTCAGTGGATGTTGCCTTTACTTCT AGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATT GTACCCGCGGCCGATCCACCGGTCCGGAATTCCCGGGATATCGTC GACCCACGCGTCCGGGCCCCACGCTGCGCACCCGCGGGTTTGCTHuman PGK GGGTTGCGCCTTTTCCAAGGCAGCCCTGGGTTTGCGCAGGGACGC 13 Promoter GGCTGCTCTGGGCGTGGTTCCGGGAAACGCAGCGGCGCCGACCCT GGGTCTCGCACATTCTTCACGTCCGTTCGCAGCGTCACCCGGATC TTCGCCGCTACCCTTGTGGGCCCCCCGGCGACGCTTCCTGCTCCGC CCCTAAGTCGGGAAGGTTCCTTGCGGTTCGCGGCGTGCCGGACGT GACAAACGGAAGCCGCACGTCTCACTAGTACCCTCGCAGACGGA CAGCGCCAGGGAGCAATGGCAGCGCGCCGACCGCGATGGGCTGT GGCCAATAGCGGCTGCTCAGCAGGGCGCGCCGAGAGCAGCGGCC GGGAAGGGGCGGT GCGGGAGGCGGGGT GT GGGGCGGTAGT GT GG GCCCTGTTCCTGCCCGCGCGGTGTTCCGCATTCTGCAAGCCTCCG GAGCGCACGTCGGCAGTCGGCTCCCTCGTTGACCGAATCACCGAC CTCTCTCCCCAGGHuman G6Pase GTGGGCCGTCTAGACTCTGTCCTGTGTCTCTGGCCTGGTTTCGGGG 14 Promoter ACCAGGAGGGCAGACCCTTGCACTGCCAAGAAGCATGCCAAAGT TAATCATTGGCCCTGCTGAGTACATGGCCGATCAGGCTGTTTTTGT GTGCCTGTTTTTCTATTTTACGTAAATCACCCTGAACATGTTTGCA TCAACCTACTGGTGATGCACCTTTGATCAATACATTTTAGACAAA CGTGGTTTTTGAGTCCAAAGATCAGGGCTGGGTTGACCTGAATACT'CTGAT ACA GGGCATAT AAAAC AGGGGCA AGGCA CAGACTC ATA G CAGAGCAATCACCACCAAGCCTGGAATAACTGCAAGGGCTCTGC TGACATCTTCCTGAGGTGCCAAGGAAAGGGAGGAAGGAATTCTC CACTTTGGGATCCAGTCAACMinimal GAACCCCTTGAGGACAGAAATGACAAGAAGACCCCCGCGGGACT 15 Mouse PKD2 TGACGAGGAGGTGGATGAGGACATCCTATGAGAAGCGTCAGAAG Promoter TGAGGGCAGAGATGCACAGATCTTGGCCAGGAATTTAACTTTTAAA AG AG A I GGGAA A ACC A FGGAAAATCGAAT I TGGAG FAGGOT T F GCCTGACCCAACTGCTGTCTTAAGTTGGCTGTGCCAGCTGGGGCGTAGCGAGGATGGAGATCTAGACCTAACGCCTGAGGCAGGATTCTAttorney Docket No. TORQ-012 / 02WO 339010-2071Description Amino Acid or Nucleotide Sequence SEQ ID NO GTTACTCAAGGGCTGGCCAGGAGCGGAAATGAGGTAATGGTGGA GGCATCAGAAAGTTCCAAAATCCAACCCTTGGATGCCTCTAGATG TCTGGAGAGTGGAGACAGGAAATCATCCAGTAAACACAGAATCA AGGAGCTGAGGGTCACCAAATGCAAGCCTTGCCGCCTTGGTGATTTCTTTCTGGTTTTTAAACGAAGGGTGGATTTCTTTGCCTTCCCTGAGAATTCAGTTCAAGCTGGATTTCACTAGTTCTTTCTATCCCAACAA AAGATCTCCATCTTTACCCTCCTCCTTGCACCTTTGCAAACCCAGG GGGAGTATGGACAGAGTCTAAGCACTATCTTAAGGTCATGCAGG CATGGACACACCCAGGAGCCCCTTTCCCGACGAGCTGGCTGACCA AGCAAGCTACCCCGTAGGAATGTGGGCACCGATGGGGACCCTGG CTCTACTTGGATCCCTGGCCGCTGGGGCCTTTTCGACGCGCCTAA GTCCAGTCCAGGCGAGGCCAGGGCCCGGGAGGCGGGCCGAGGGG GCGGGCGGAGGCGGGGAGCAGCAGGCCGACTCCGGGAAGAAAA GAACGCTCCTGCGGCAGAGGGCGGCGGCACTCCCGGGCGCGCTG GGCGCCAGTGAGCGCCGTTAACTCC WT AAV25’ TTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGG 16 ITR - Flip CGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCA GTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCAC TAGGGGTTCCT WT AAV23’ AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTC 17 ITR - Flop GCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCG GGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGA GGGAGTGGCCAABovine GHpA CTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGT 18 GCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAA TAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCT ATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTG GGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGG TGGSGG- ACCGGCGGCAGCGGCGGCTACCCATACGATGTTCCAGATTACGCT 19 3XHA TACCCATACGATGTTCCAGATTACGCTTACCCATACGATGTTCCA GATTACGCTKozak GCCGCCACCATGG 20 SequenceExample 2 — In vitro Expression of PKD2 Cassettes via Transfection

[0288] This Example aims to show that the PKD2 cassettes designed based on Example 1 can be used to express PKD2 mRNA and PC2 protein.

[0289] The PKD2 cassettes were subcloned into expression plasmids. PAz / 2-knockout mouse inner medullary collecting duct cells (1MCD3 cells) were transfected with the recombinant plasmids, harvested 24 hours after the transfection, and levels of PKD2 RNA and PC2 protein were assessed using reverse transcription digital PCR (RT-dPCR) and capillary electrophoresis-based immunodetection, respectively.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0290] As shown in Figs. 2A and 2B, the CBA promoter drove the highest expression of PKD2 mRNA relative to the hPGK, mPKD2, and hG6Pase promoters. Notably, the CBA promoter drove expression of PKD2 mRNA to a level similar to that of GAPDH mRNA (Fig. 2B). Relative quantity of PC2 protein was then assessed via immuno-detection of HA tag signal on a capillary¬ based immunoassay system. Signals were quantitated as the area under the curve (AUC) of peaks around 116 kD and normalized to the CBA-PKD2-FL signal. As shown in Fig.2C, PA<72-knockout immortalized mouse collecting duct cells (IMCD3) cells transfected with CBA, hPGK, or mPKD2-driven PKD2 cassettes expressed varying levels of PC2 protein. The hG6Pase-driven PKD2 cassettes did not express any detectable PC2 protein (Fig. 2C) despite expressing PKD2 mRNA at a level similar to that of hPGK-driven cassettes (Fig. 2A).

[0291] To determine if PC2 protein expression would be affected by the type of host cell, human HEK293T cells and mouse Pkd2 -knockout IMCD3 were transfected with plasmids comprising CBA-driven PKD2 cassettes. Relative quantity of PC2 protein was then assessed via immunodetection of HA tag signal on a capillary-based immunoassay system. As shown in Fig.3A, the human PKD2-FL coding sequence was expressed at the highest level in human HEK293T cells, and there was no difference in PC2 protein expression among the engineered PKD2-FL; CO, PKD2-FL; ACpG, and PKD2-FL; C(); ACpG coding sequences. / Xs shown in Fig. 3B, the PKD2-FL; CO coding sequence was expressed at the highest level in mouse PAa?2-knockout IMCD3 cells, followed by the PKD2-FL; CO; ACpG, PKD2-FL, and PKD2-FL; ACpG coding sequences.

[0292] Together, these data demonstrate that PKD2 cassettes containing a CBA, hPGK, or mPKD2 promoter can support PC2 protein expression in mouse and human cell lines in vitro following transfection.Example 3 — In Vitro Expression of PKD2 Cassettes via AAV Transduction

[0293] This Example aims to show that the PKD2 cassettes designed based on Example 1 can be expressed in vitro after delivery by recombinant AAV viruses.

[0294] Recombinant AAV.k20-packaged PKD2 cassetes were prepared using conventional methods. Mouse PAz / 2-knockout IMCD3 were transduced with the recombinant AAV.k20 viruses at different multiplicities of infection (MOls) and were harvested 72 hours later for PKD2 mRN A and PC2 protein expression analysis by RT-dPCR and capillary electrophoresis- based immunodetection, respectively.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0295] As shown in Figs. 4A and 4B, the CBA promoter drove the highest expression of PKD2 mRNA relative to the hPGK, mPKD2, and hG6Pase promoters in cells transduced with AAV.k20-packaged PKD2 cassettes. As shown in Figs. 4C and 4D, the CBA promoter drove the highest expression of the PC2 protein in a dose-dependent manner in cells transduced with AAV.k20-packaged PKD2 cassettes. Notably, the engineered PKD2-FL; CO, PKD2-FL; ACpG, and PKD2-FL; COiACpG coding sequences expressed higher levels of PC2 protein compared to the PKD2-FL coding sequence under control of the CBA promoter (Figs. 4C-4D). The mPKD2- and hG6Pase-driven PKD2 cassettes did not express detectable amounts of PC2 protein in transduced cells (Figs. 4C-4D). Transduced cells with detectable PC2 protein expression showed a strong positive correlation (R2= 0,82) between PC2 protein expression and PKD2 mRNA expression (Fig. 4E).

[0296] Single cell immunocytochemistry analysis was performed in FAt / 2-knockout IMCD3 cells transduced with AAV.k20-packaged PKD2 cassettes. Fig. 5A shows a strong dose-dependent increase in the proportion of Pfo / 2-knockout IMCD3 cells expressing PC2 protein. CBA-driven PKD2 cassettes exhibited the highest level of PC2 protein expression followed by hPGK-driven and mPKD2-driven PKD2 cassettes (Fig. 5A). Fig. 5B shows a dose-dependent increase in the intensity of PC2 protein expression (measured by HA signal) in transduced cells. It is noteworthy that the increased proportion of cells expressing PC2 protein was concurrent with the increased intensity of PC2 protein in transduced cells, indicating a dose-dependent increase in the abundance of PC2 protein. hG6Pase-dnven PKD2 cassettes did not express detectable amounts of PC2 protein in Ffc72-knockout IMCD3 cells across all MOIs (Fig. 5A and Fig. 5B).

[0297] These findings suggest that the following promoters drive PC2 protein expression from highest to lowest: CBA > hPGK »> G6Pase or mPKD2. Taken together, these data demonstrate that recombinant AAV.k20-packaged PKD2 cassettes driven by a CBA or a hPGK promoter can support PC2 protein expression.Example 4 — In Vitro Evaluation of Additional PKD2 Cassette Designs

[0298] This Example aims to evaluate how different promoters modulate PKD2 mRNA and PC2 protein expression following transfection with PKD2 cassettes.

[0299] As shown in Fig. 6A, additional PKD2 cassettes were designed to modulate PC2 expression via three mam strategies. As a first strategy, PKD2 cassettes comprising the ubiquitous promoters CBA and hPGK were designed to decrease liver expression by incorporating two miR-Attorney Docket No. TORQ-012 / 02WO 339010-2071122 binding sites downstream of the PKD2 CDS, as miR-122 is primarily expressed in the liver at high levels. The goal of this evaluation was to ensure that miR-122-containing cassettes expressed PKD2 mRNA and PC2 protein similar to their cassette counterparts lacking miR-122 binding sites in cell culture models where miR-122 is not expected to be highly abundant (i.e., PAD2-knockout HEK293 and Pfc72-knockout IMCD3 cells). As a second strategy, promoters expected to be active in the kidney were used to direct and increase PKD2 / PC2 expression in the kidney. Finally, as a third strategy, mini-PKD2 promoters derived from human, pig, and mouse species were used to recapitulate endogenous PKD2 / PC2 expression. Ail PKD2 cassettes comprised a promoter, a Kozak sequence, a codon-optimized / CpG-depleted PKD2 (PKD2-FL; CO; ACpG) or a wild-type, full-length PKD2 (PKD2-FL) coding sequence, a sequence encoding a 3X HA -tag, and a bGHpA or a rBGHpA signal. The cassettes were flanked by wild-type A AV2 inverted terminal repeats (ITRs, 145 bp) m a plasmid backbone. The plasmids were transfected into mouse PAv / 2-knockout IMCD3 cells and human PXD2-knockout HEK293 cells, cells were harvested 24 hours later, and RNA and protein expression were analyzed by RT-dPCR and a capillary-based immunoassay system, respectively.

[0300] Additional sequences related to the PKD2 cassettes are listed in Table 4 below.Table 4.Description Amino Acid or Nucleotide Sequence SEQ ID NO EIFla GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGT 21CCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGC CTAGAGAAGGT GGCGCGGGGTAAACT GGGAAAGT GAT GT CGT GTACTGGC FCCGCC I T 1 TTCCCGAGGGTGGGGGAGAACCGT AT ATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGT TTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGG GCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTA CTTCCACGCCCCTGGCTGCAGTACGTGATTCTTGATCCCGAGCT TCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAA GGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCTTGGGC GCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTG TCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGAT GACCTGCTGCGACGCTTTTTTTCTGGC’AAGATAGTCTTGTAAAT GCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTGGGGCCG CGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGC GAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGG GTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCG CCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGT CGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAAttorney Docket No. TORQ-012 / 02WO 339010-2071GCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCC GTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTACCGGGCG CCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGTC GTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCC C ACA CTGA GTGGGTGGA GA CTG A AG'FTA GGCC AGCTTGGCACT TGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAGTTTGGATCTT GGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTT CCATTTCAGGTGTCGTGAmECAD ACCTGACATCCTCACACCAATACACATAAACAAAATAAATTAT 22TAAAAAATAAAACCCAACAGAACAACTGATGAGGAGGGCTGG AGAGATGGCTCAGTGGTTGAGAGCACTGGCTGCTTCCAAAGGA CCCAGGTTCAAATCCCAGCAACCACATGGTAGTTACAGCTGTC TGTAATTCCAGTTCCAGGAGATCTGATACCCTCACATAGACAT AC ACGGAGGGAG A ACA ATG FAAGT A A A AGA FTT AAAA’T A A A A AGCAACAACAACAAAACAAAACAAAACAAAAAACCTGATGGA TGTGGGATGCATAGGATGCTGCTACTTGGGGGTCCCAAGGTCA AAGCCAGTCTGGGTTACCTAGTCCTGAGAACTGAAGTTAAGAG AACAGATTTTAGCCGGGCAGTAAGGTAGTGGGTACCTGTAGCT CCATCCCAGAAGTGAGAAGGCTGAGTTTGAACAATGATGAGTT TAAAGCCATGCTGGGCTACATAGCAAGGCTATGTCTCAAAAGG GAGCCGGTCAGGTGTTAGAAAATTAGATTAGCCTGGTCTGGTA CCCCACTTGTGCAATCCCAGCATTCGGGAGACTGAAACAGGAG GATGGCTAAGACTAAGACAATTCCAGGCCCACCTGAGTTGGAA AACAAACAAACAAAAAAGCTACCAAACAAAATAAAACCGTCG GAGAAATAGCTCAGTCAGTAAAGGCCAATGGCGGCAATGCAA TCCCAAGACCCTCTTGGTGGAAGAAGAGAATTGACTCTTGAAG GCTGTCGTCTTATCTCCACAATCGGTCTGTGGCACGTGCGTTTG CGAGCACAGGATCGCGCTCTCGCCCGCGCACGCACCCCTCCCC CCATGTTAAAATGTCATTTAAAATCCCTAAGCAAACAAACTCA TCCAACCAAAGAAAATAAAAACATAAGAAACAAAACGGAAAC CTAGATGATGAATAAAGTCCTTTGTAACTCCATGTCTCCGTGG GTCAGAGCACAGCTAGGCTAGGATTCGAACGACCGTGGAATA GGAAGCTGGGAAGTCTTCTAAGGCCGGCCCCATGCCACCAACT ACAGACAGGGGTGGAGGAAGTTGAGGGCCCTGCAGTTCCTTG GCTGCCACCTGCAGGTGCGTCCCCACTCCAATCAGCGGCCTCCGG GGGCGGTGCCTGCGGGCTCACCTGGCGGCCGCAGCCTCTGCGMini- CAAAAATTAAACAAATTCAGGAACTGAAAAACAATAATTAAT 23 hWNKl-RP AGAATATGTAAGCATATCATATATATCACAGTATTATAGATTTT CCTTTAACCAATCATGTGTGTATTTTAAAGATACTGAAAGGAA AATGTAGTCTTTTCTATTTAACATTTTTAGTGCTCTTTTTTTCCT GAATATTTCCAGATAAAATAGGCATTGGAACTCAATCCCCAGA TGTATCTTTTCTACAGTGGGTAGCACCAGAGGTCTCTGTTCAGT'rC'rTTTAGCATACTTGGGCTGCTTCTCTAATCTTTCCTACATTCAT GTATTTCAGAGGTCAACTAGAGATTTGGGCAGAGTGGATACAC AGAATTTAGGGCTCTCCTTTGGTGCCTGCTTTCTGAGATTTCCT CCTTTCCTTTCTAGATCTGTGCTGTCCAGTAGAACTTTCTGCCA TTATGGAAATGTTGTATAACGTACACCATCCAGAATGGTAGCC ATTTGCCTCATGTGGCTGTTCAGCACGTGAAATGTGGCTAGTG CAACTGAGAAATGAAATTTTTTATTTTAGTTAATCTACATTTGA GTAGTCACATTTGGCTAGTGGCAACTTTATTGGACAGACCATCCTTGCAGAAAGGTCTGTTAGACAGCACTGTTCTTGATGCTGTGAttorney Docket No. TORQ-012 / 02WO 339010-2071GTCATTCCAAACTCAGTCGCCATTTCTGTCTGGGGTTTAGCAGA CCTGCAGAATCAGAACAGGCCCTTTCCTTAGGTAAAAATGTAT TATAAAGAAGGAAACTCAAGTAGTGCTTTCCCTTTTTCCCTTCC AACTATGACTGATTTTGATTGCTCTCCAGAGCCTTCAAGTAATA GTTGTTTTGTACTTCTCACCCCGGTGAGTAACGTCTGAAGCTTC TGCTCGCATTGAGTCTGAATCGTTCTTTTAATTTAAGGTACCCC AGAGTTCmKSPC AGCTTGCTCTGCCATGGGAAGGTCCCCAAACCTGAAAAAGAAA 24CCCAGGCGCCTTGGGCAAACGGCTTAGCCTCTCTGTACCCCAG AGGGGTATGACAAGGGGACAGTTCCCTCTTCAGAATGAGTGGG TGGGATAGCAGCGATATGTGCTCACAGAAGTTCTGGGTGCTGG TTAGTAGGCACTCAGTATGTGTGAGACCTCCTTCAACAACCCC ACATATAGCACTAACCTAGGCTCAGTGCCTCAGTGATCCTAAA ACAGACACTCAGCTTGTCCCAGTCCCGCTTCCATCTGGAATGCT GACCAGATATACCATGAGCAGTCCTCGGGGGAGACCCCAAAA CATGGAAGGAAAGTGAGAGGAGGGAGACCAGGGTCTTCATAC TCTGTACCCTCATCTGAGATTCCCAGAAAACAAGGTTTCTGCTG GGTCTGCCTTTCAGGCCTTACAGGGGGAGGTGAGGAGGCCCTG GTCAGCCCTGTAACTCCCTCAAAGTCACCTGAGCTGGCTCGGT CTTCTCTTTTCTGCTTAGAGACATGAGACAGGTCAGAGCCTCAC CCTTATCTGTGTTCAGGAGACCATGTGAGGCTAGGAGAGCTTC AGGGGGACCTCTAGGCTTCTGTCCCACCCACTGGTCTCACTGA TGAATGTTCTAGAGCTCCGAACACACACTTGGGGTCAGCTGCC CTTTGGGTTACACTCCCAGTCCTTTCCTTAGCTCCACGGGCCAC TTGGGGCAGGACAAGGTCATAGCCTACTCAGAGGAACTCCGA AGCTAATACGTGAGAAACAAGCTGGTGTCTTCTGGGCATCCGT AGAGGCAGTTCATGGTATGGAGTGGGGGAGCCTAATCCAGCCT GTGAATGTAAGGGCTTTCCTGAAGAAGGGGTTATTAGTGCTTG TTGCTAAAGCGTAAGCAGGTACTTACTGTCCTGTAGAAAGGTG GGAAGAGCATTCCAGGCACAAGGAACAATATCTGGAGAGATC TGGCTACAGATGCCACTGCACAGAGGAGAAGTGGGAGCCAAG TCTGAACACACACACACACACACACACACACACACACAGAAG TTATGTCTAAGCCCAGAGGGCCAGCAAGTGCTCATTGGGCTGT GTCAAGGGGGCAGTGACAGACCAAGAGCTGCCCACCTCCTAG GGCTGGCAGTCACGGATGCTGAGCAGATCTGGCTCTCCAAAGT CAATAAGTAACTTGGGGGGACTAGGCGGGGCCAGGCCTGCTCC TGTGGGCCCCGGTGGCATTTTCCACTCCTGAGCAAGCACGGCC AGACCGCCTACCTGCTCAAGTGTCCACCTTGCCTCGCCCCACCT AAGCCAAATTTGC RP-hPGK CAAAAATTAAACAAATTCAGGAACTGAAAAACAATAATTAAT 25AGAATATGTAAGCATATCATATATATCACAGTATTATAGATTTT CCTTTAACCAATCATGTGTGTATTTTAAAGATACTGAAAGGAA AATGTAGTCTTTTCTATTTAACATTTTTAGTGCTCTTTTTTTCCT GAATATTTCCAGATAAAATAGGCATTGGAACTCAATCCCCAGA TGTATCTTTTCTACAGTGGGTAGCACCAGAGGTCTCTGTTCAGT TCTTTTAGCATACTTGGGCTGCTTGGAATCTTTCCTACATTCAT GTATTTCAGAGGTCAACTAGAGATTTGGGCAGAGTGGATACAC AGAATTTAGGGCTCTCCTTTGGTGCCTGCTTTCTGAGATTTCCT CCTTTCCTTTCTAGATCTGTGCTGTCCAGTAGAACTTTCTGCCA TTATGGAAATGTTGTATAACGTACACCATCCAGAATGGTAGCCATTTGCCTCATGTGGCTGTTCAGCACGTGAAATGTGGCTAGTGAttorney Docket No. TORQ-012 / 02WO 339010-2071CAACTGAGAAATGAAATTTTTTATTTTAGTTAATCTACATTTGA GTAGTCACATTTGGCTAGTGGCAACTTTATTGGACAGACCATC CTTGCAGAAAGGTCTGTTAGACAGCACTGTTCTTGATGCTGTG GTCATTCCAAACTCAGTCGCCATTTCTGTCTGGGGTTTAGCAGA CCTGCAGAATCAGAACAGGCCCTTTCCTTAGGTAAAAATGTAT TATAAAGAAGGAAACTCAAGTAGTGCTTTCCCTTTTTCCCTTCC AACTATGACTGATTTTGATTGCTCTCCAGAGCCTTCAAGTAATA GTTGTTTTGTAGGGTTGCGCCTTTTCCAAGGCAGCCCTGGGTTT GCGCAGGGACGCGGCTGCTCTGGGCGTGGTTCCGGGAAACGC AGCGGCGCCGACCCTGGGTCTCGCACATTCTTCACGTCCGTTC GCAGCGTCACCCGGATCTTCGCCGCTACCCTTGTGGGCCCCCC GGCGACGCTTCCTGCTCCGCCCCTAAGTCGGGAAGGTTCCTTG CGGTTCGCGGCGTGCCGGACGTGACAAACGGAAGCCGCACGT CTCACTAGTACCCTCGCAGACGGACAGCGCCAGGGAGCAATG GCAGCGCGCCGACCGCGAT GGGCTGT GGCCAATAGCGGCT GCT CAGCAGGGCGCGCCGAGAGCAGCGGCCGGGAAGGGGCGGT GC GGGAGGCGGGGTGTGGGGCGGTAGTGTGGGCCCTGTTCCTGCC CGCGCGGTGTTCCGCATTCTGCAAGCCTCCGGAGCGCACGTCG GCAGTCGGCTCCCTCGTTGACCGAATCACCGACCTCTCTCCCCA GGmPAX8 CAGGGGAAGAGAAGGGTTGAAGGTTGTCATAGAGTGACCTTG 26AGCTCACTCTGGTGGTCAGCGATCCAAGAGTTTCCAGAGAACC ACATCCCCAGACCCCAGGCTTTCATCTCCCACTCCCCAGTACTT CCTGGCCTGCTGAGTTAGGATTCTGTGGCCGCCCTCTCTTCCCT CCTCCAGAAACAGAAGCTCCAGCGAATGTTTCTAACTCCTGAG TCCACTCAGCCATGTCCCCCTTGCAAAATCCCCCCCTTCCCTCA CCCCCCTGCACCCAGCCAGCCAATCCGAGGCCTGAGAGCCTCG CCAGACACATCTCCCAGGGCCTCGGCGGGATAAAACTGGCTGG CGATGCCAGGTGGATGGGAGCAAACTTCAGAAGGAAGAGACG CCTGGGCCTTGGGCA CCCTC AGGGGCA GA CCC AGGCA GA A A G GGCCTGAGGCCAGCCGGCCAGGTATGTCACCTAGGGGCTAGA AGGGAGCTGGAAACAGCTGGGTAAGAAGACTAGCAGCCTTTTT CCTCTCAAGTTAAGTGGGATGAGGTAGTATCTATGGAGGGACA GGGCTGGAGTTTGGGGCTTCCTTGGTGCACCCCTAACCGTGGA TCCTCCCAACAGGCACATTTGCTAGGCCAGGCTGAGGTTGGCC TCACTTCACCCCACTCTGGACACTCAGTAGGAGTCAAAAGACC TGGCTTG G G G AGGGG ATGTG G G TTTGAGGCCTCG ATTCTCATG CCCTTCGCCCTGGGTCTATATGCAGGGTAGCTGCGTGGCAGCC AGAGCTGCCAGGACCTGCGTAGGAAAGCTGCGAGTGTCCCTCA GTCTGTGAGCGACTCCCCGGCGhPKD2 AGGAGGCCCGCCAAATGCACAGCAAGTCAGCCTTGGTAGTTTG 27CAAATGGAAAGATTTCTTTGACCCGCTGGAGACTTCAGCTCAG GCTGGATGTCGCTAATTCACACCATCTCACCAAAAGGTTCTTTC CCTGCCCTTTGGTGAATCGAGGCAGAAGTGGCTGGTGGATAAC TGGGACCCAAGAGTTATCTCAGCGTCGGGCAGGCATCGACAGC TCCAGGAGCCCTTTCCCTGCAGGCGGGCTGGCGGGTGAGCGAT TCCCTCCCTTCCCAGGCCTTCCCGGGAATGGAACGTGGGCGCC CGCGCCCGGGCTCCGGTTCCACTTGGAACGCGGACTCGGGAGC CGCCGGGCGAGCGCGCAGACCGCGGGGGCGGCTCCCTCGCAC CTCCCCGCTCAGCGCGCCGCGGCGGATGGGCGAGGCGGGGCGAGGCCAGAGGGAGGCGGGCCAAAGGGGCGGGCGAGAGGACGAttorney Docket No. TORQ-012 / 02WO 339010-2071CGGGACCGCGGGGAGGTCGGGGGCGGGGAGCAGGCGGCGGCG GGCGCCGGGAAGAAAGGAACATGGCTCCTGAGGCGCACAGCG CCGAGCGCGGCGCCGCGCACCCGCGCGCCGGACGCCAGTGAC CGCGhPKD2-G4ml AGGAGGCCCGCCAAATGCACAGCAAGTCAGCCTTGGTAGTTTG 28CAAATGGAAAGATTTCTTTGACCCGCTGGAGACTTCAGCTCAG GCTGGATGTCGCTAATTCACACCATCTCACCAAAAGGTTCTTTC CCTGCCCTTTGGTGAATCGAGGCAGAAGTGGCTGGTGGATAAC TGGGACCCAAGAGTTATCTCAGCGTCGGGCAGGCATCGACAGC TCCAGGAGCCCTTTCCCTGCAGGCGGGCTGGCGGGTGAGCGAT TCCCTCCCTTCCCAGGCCTTCCCGGGAATGGAACGTGGGCGCC CGCGCCCGGGCTCCGGTTCCACTTGGAACGCGGACTCGGGAGC CGCCGGGCGAGCGCGCAGACCGCGGGGGCGGCTCCCTCGCAC CTCCCCGCTCAGCGCGCCGCGGCGGATGGGCGAGGCGGGGCG AGGCCAGAGGGAGGCGTGCCAAAGGGGCGGGCGAGATGACGC GTGACCGCGGGGAGGTCGGGGGCGGGGAGCAGGCGGCGGCGG GCGCCGGGAAGAAAGGAACATGGCTCCTGAGGCGCACAGCGC CGAGCGCGGCGCCGCGCACCCGCGCGCCGGACGCCAGTGACC GCGpPKD2 AGGGGCTAGTTGGTCGTTTTCAAGTCAAAGCTTTCCTTTGCCTC 29CGCTGCAGACCAGCGCAGGCTGGGTTTCGCCTGTTCGCACCAC CGTCCCCTAAAATGGTTCACCCCTCTCCCCGCACTTTGGTAAAT CGAGGCAGAAGTGGCTGGGTGGGGGGAACGGAGACCCAAGCC TTATTTCAGGGTCGCGCAGCCAGGGACAGCTCCAGAAGCCCTT GGCTCGCGGGTGGACGGACGGCTCGGCGAGTCCCTCCCCGCCC GAGCCCTTCCACCCGAAAGGAATGCGGCCGCCCGCGCCCGGGC TCAGGCTGCACTTGGAACGCGGACCCGGGCGCCGCGCGCCGA GCGCGCGGACCACGGGGGCGGCTGCCTTGCGCCTCCCCCTGCA GCGCGCGGCGGCCGCCGGGCGAGGCGAGGCGAGGGGGAGGCG GGCCGAGGGGGCGGGCGAGGGACGCGGGACCGCGGGGAGGTC GGGGGCGGGGAGCAGGAGCCGCGGGCCCCGGGAAGAAAAGA ACATGGCTCCTGCAGCGGGCAGCGCCGAGCGGGGCACCGGGC GAGGGCGCGCCTGCAGCCAGTGACCGCGpPKD2-G4ml AGGGGCTAGTTGGTCGTTTTCAAGTCAAAGCTTTCCTTTGCCTC 30CGCTGCAGACCAGCGCAGGCTGGGTTTCGCCTGTTCGCACCAC CGTCCCCTAAAATGGTTCACCCCTCTCCCCGCACTTTGGTAAAT CGAGGCAGAAGTGGCTGGGTGGGGGGAACGGAGACCCAAGCC TTATTTCAGGGTCGCGCAGCCAGGGACAGCTCCAGAAGCCCTT GGCTCGCGGGTGGACGGACGGCTCGGCGAGTCCCTCCCCGCCC GAGCCCTTCCACCCGAAAGGAATGCGGCCGCCCGCGCCCGGGC TCAGGCTGCACTTGGAACGCGGACCCGGGCGCCGCGCGCCGA GCGCGCGGACCACGGGGGCGGCTGCCTTGCGCCTCCCCCTGCA GCGCGCGGCGGCCGCCGGGCGAGGCGAGGCGAGGGGGAGGCG TGCCGAGGGGGCGGGCGAGTGACGCGTGACCGCGGGGAGGTC GGGGGCGGGGAGCAGGAGCCGCGGGCCCCGGGAAGAAAAGA ACATGGCTCCTGCAGCGGGCAGCGCCGAGCGGGGCACCGGGC GAGGGCGCGCCTGCAGCCAGTGACCGCGmPKD2-483 TTCTGTTACTCAAGGGCTGGCCAGGAGCGGAAATGAGGTAATG 31GTGGAGGCATCAGAAAGTTCCAAAATCCAACCCTTGGATGCCTCTAGATGTCTGGAGAGTGGAGACAGGAAATCATCCAGTAAACAttorney Docket No. TORQ-012 / 02WO 339010-2071ACAGAATCAAGGAGCTGAGGGTCACCAAATGCAAGCCTTGCC GCCTTGGTGATTTCTTTCTGGTTTTTAAACGAAGGGTGGATTTC TTTGCCTTCCCTGAGAATTCAGTTCAAGCTGGATTTCACTAGTT CTTTCTATCCCAACAAAAGATCTCCATCTTTACCCTCCTCCTTG CACCTTTGCAAACCCAGGGGGAGTATGGACAGAGTCTAAGCAC TATCTTAAGGTCATGCAGGCATGGACACACCCAGGAGCCCCTT TCCCGACGAGCTGGCTGACCAAGCAAGCTACCCCGTAGGAATG TGGGCACCGATGGGGACCCTGGCTCTACTTGGATCCCTGGCCG CTGGGGCCTTTTCGACGCGCCTAAGTCCAGTCCAGGCGAGGCC AGGGCCCGGGAGGCGGGCCGAGGGGGCGGGCGGAGGCGGGG AGCAGCAGGCCGACTCCGGGAAGAAAAGAACATGGCTCCTGC GGCAGAGGGCGGCGGCACTCCCGGGCGCGCTGGGCGCCAGTG AGCGCCmPKD2-389 CT GGAGAGTGG AG ACA GGA A AT CATCCAGT AAACACA GA A T C 32AAGGAGCTGAGGGTCACCAAATGCAAGCCTTGCCGCCTTGGTG ATTTCTTTCTGGTTTTTAAACGAAGGGTGGATTTCTTTGCCTTC CCTGAGAATTCAGTTCAAGCTGGATTTCACTAGTTCTTTCTATC CCAACAAAAGATCTCCATCTTTACCCTCCTCCTTGCACCTTTGC AAACCCAGGGGGAGTATGGACAGAGTCTAAGCACTATCTTAA GGTCATGCAGGCATGGACACACCCAGGAGCCCCTTTCCCGACG AGCTGGCTGACCAAGCAAGCTACCCCGTAGGAATGTGGGCACC GATGGGGACCCTGGCTCTACTTGGATCCCTGGCCGCTGGGGCC TTTTCGACGCGCCTAAGTCCAGTCCAGGCGAGGCCAGGGCCCG GGAGGCGGGCCGAGGGGGCGGGCGGAGGCGGGGAGCAGCAG GCCGACTCCGGGAAGAAAAGAACATGGCTCCTGCGGCAGAGG GCGGCGGCACTCCCGGGCGCGCTGGGCGCCAGTGAGCGCCrBGHpA AATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTT 33TTGTGTCTCTCARenal CAAAAATTAAACAAATTCAGGAACTGAAAAACAATAATTAAT 161 Enhancer AGAATATGTAAGCATATCATATATATCACAGTATTATAGATTTT Element (RP) CCTTTAACCAATCATGTGTGTATTTTAAAGATACTGAAAGGAA AATGTAGTCTTTTCTATTTAACATTTTTAGTGCTCTTTTTTTCCT GAATATTTCCAGATAAAATAGGCATTGGAACTCAATCCCCAGA TGTATCTTTTCTACAGTGGGTAGCACCAGAGGTCTCTGTTCAGT'rC'rTTTAGCATACTTGGGCTGCTTGGAATCTTTCCTACATTCAT GTATTTCAGAGGTCAACTAGAGATTTGGGCAGAGTGGATACAC AGAATTTAGGGCTCTCCTTTGGTGCCTGCTTTCTGAGATTTCCT CCTTTCCTTTCTAGATCTGTGCTGTCCAGTAGAACTTTCTGCCA TTATGGAAATGTTGTATAACGTACACCATCCAGAATGGTAGCC ATTTGCCTCATGTGGCTGTTCAGCACGTGAAATGTGGCTAGTG CAACTGAGAAATGAAATTTTTTATTTTAGTTAATCTACATTTGA GTAGTCACATTTGGCTAGTGGCAACTTTATTGGACAGACCATC CTTGCAGAAAGGTCTGTTAGACAGCACTGTTCTTGATGCTGTG GTCATTCCAAACTCAGTCGCCATTTCTGTCTGGGGTTTAGCAGA CCTGCAGAATCAGAACAGGCCCTTTCCTTAGGTAAAAATGTAT TATAAAGAAGGAAACTCAAGTAGTGCTTTCCCTTTTTCCCTTCC AACTATGACTGATTTTGATTGCTCTCCAGAGCCTTCAAGTAATAGTTGTTTTGTAAttorney Docket No. TORQ-012 / 02WO 339010-2071

[0301] As shown in Fig. 6B, all PKD2 cassettes expressed PKD2 mRNA above background in transfected Pfc / 2-knockout IMCD3 cells. Mock-transfected cells showed less than ~102RNA copies / ng (data not shown). In PKD2-knockout HEK293 cells, most cassettes expressed PKD2 mRNA at similar or greater levels than hPGK following transfection (Fig. 6C). Mock-transfected cells showed less than 36 RNA copies / ng (data not shown). PC2 protein was detectable for a subset of the cassettes tested (Fig. 6D). Transfected cells with detectable PC2 protein expression showed a strong positive correlation between PC2 protein and PKD2 mRNA that was driven by the promoters clustering into high and moderate expressors (Fig. 6E). Fig. 6F provides a summary table listing the PKD2 cassettes in which PC2 protein was detectable or PC2 protein was not detectable.

[0302] Together, these results demonstrate that PKD2 mRNA can be expressed from a wide variety of promoters in both mouse and human-derived cell lines, PC2 protein is detectable when the PKD2 coding sequence is preceded by a medium-strength or strong, ubiquitous promoter. Additionally, an hPGK promoter with an upstream renal enhancer element (RP-hPGK) and a modified, mini human PKD2 promoter (mini-hPKD2) also supported detectable PC2 protein expression.Example 5 — In Vitro Evaluation of UTRs to Modulate Expression of PC2

[0303] This Example aims to evaluate how PKD2 cassettes containing different 5’ and 3’ untranslated regions (UTRs) affect PKD2 mRNA and PC2 protein expression following transfection.

[0304] PKD2 cassettes were designed with naturally occurring UTR sequences or synthetic sequences integrated upstream of the Kozak sequence (5’ UTR) or downstream (3 ’U TR) of the PKD2-FL; CO; ACpG coding sequence (Fig. 7A).

[0305] The sequences related to these PKD2 cassette designs are provided in Table 5 below.Table 5.Description Amino Acid or Nucleotide Sequence SEQ ID NO5’ UTR 9765 CTTGTCTCGCTCCGGGGAACGCTCGGAAACTCCC 34GGCCGCCGCCACCCGCGTCTGTTCTGTTACACAA GGGAAGAAAAGCCGCTGCCGCACTCCGAGTGT5" UTR 11674 CACTCGCGCTGCCATCACTCTTCCGCCGTCTTCG 35CCGCCATCCTCGGCGCGACTCGCTTCTTTCGGTTCTACCAGGTAGAGTCCGCCGCCATCCTCCACCAttorney Docket No. TORQ-012 / 02WO 339010-20715’ UTR 10023 CATTCTGTGGTCTGATCATCCTGTGGTTTCGTCG 36CCGCCATCCTCGTCGCGACACGCTGTTTTCGGTT CTCGGCCCGACGAGCCATCGCCATCCTACAGC5’ UTRhHBB ACATTTGCTTCTGACACAACTGTGTTCACTAGCA 37ACCTCAAACAGACACC5’ UTR PoV_pA_scrUTR GGCTCTGAAAAAAAAAAAAAACCGAGACCCAA 38GCTAGCTAGCGTTTAAACTTAAGCTAGGTACCG AGACC5’ UTR TOP CTTCCTTTTGTACATTTGCTTCTGACACAACTGTG 39TTCACTAGCAACCTCAAACAGACACC5’ UTR TMV GTATTTTTACAACAATTACCAACAACAACAAAC 40A A CAA AC A ACATTACA ATT ACT ATTTA CAATT ACA5’ UTR TEV GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGA 41AATATAAGAGCCACC5’ UTR scrUTR AGACCCAAGCTAGCTAGCGTTTAAACTTAAGCT 42AGG’I ACCGAGACC5’ UTR RpS25 AGCGAGGCTGCTGTGGTCTACACGACTCTCTGA 43GCTTCGCC5’ UTRRpL38 CTTTCCCCGTTCTCTTCGGTTCTCATCGCTGTGAG 44T GT GCT GGGCAGGT GC GGACGCCAGAGC CGAGC CCGCGTCGCC5" UTR mActB TATAAAACCCGGCGGCGCAACGCGCAGCCACTG 45GCAGCTCCTTCGTTGCCGGTCCACACCCGCCACC AGTTCGCCCC5" UTR C3 ACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCC 46CTCTGACCCTGCACTGTCCCAGCACChACTB ACCGCCGAGACCGCGTCCGCCCCGCGAGCACAG 47CCCGCCGCCAGCTCACC5’ UTR mRpll 8 a P4_mActB CTTCCTTTTGTGACTGGCGGTGAACGAGTGCGCA 48GTGCCCGCGTTATTGTTCTGCCGGGCGGACACGT GACGCGAAGCTTTATAAAACCCGGCGGCGCAAC GCGCAGCCACTGTCGAGTCGCGTCCACCCGCGA GCACAGCTTCTTTGCAGCTCCTTCGTTGCCGGTC CACACCCGCCACCAGTTCGCCCC5’ UTR P4_mActB CGCGTTATTGTTCTGCCGGGCGGACACGTGACGC 49GAAGCTTTATAAAACCCGGCGGCGCAACGCGCA GCCACTGTCGAGTCGCGTCCACCCGCGAGCACA GCTTCTITGCAGCTCCTTCGrTGCCGGrCCACAC CCGCCACCAGTTCGCCCC3’ UTRhHBB GCTCGCTTTCTTGCTGTCCAATTTCTATTAAAGG 50TTCCTTTGTTCCCTAAGTCCAACTACTAAACTGG GGGATATTATGAAGGGCCTTGAGCATCTGGATT CTGCCTAATAAAAAACATTTATTTTCATTGCAA3’ UTR 2xmiR122 CAAACACCATTGTCACACTCCAAATTCAAACAC 51CATTGTCACACTCCA3’ UTR 5xmiR122 CAAACACCATTGTCACACTCCAAATTCAAACAC 52CATTGTCACACTCCAATATCAAACACCATTGTCAAttorney Docket No. TORQ-012 / 02WO 339010-2071CACTCCATTATCAAACACCATTGTCACACTCCAA ATTCAAACACCATTGTCACACTCCA' UTR CYBA-l,5x CCTCGCCCCGGACCTGCCCTCCCGCCAGGTGCAC 53CCACCTGCAATAAATGCAGCGAAGCCGGGACCT CGCCCCGGACCTGCCCTCCCGCCAGGTGCACCC ACCTGCAATAAATGC’ UTR CYBA CCTCGCCCCGGACCTGCCCTCCCGCCAGGTGCAC 54CCACCTGCAATAAATGCAGCGAAGCCGGGA" UTR hHBAl GCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTT 55GGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCA CCCGTACCCCCGTGGTCTTTGAATAAAGTCTGAG TGGGCGGCA’ UTR hHBBx2 GCTCGCTTTCTTGCTGTCCAATTTCTATTAAAGG 56TTCCTTTGTTCCCTAAGTCCAACTACTAAACTGG GGGATATTATGAAGGGCCTTGAGCATCTGGATT CTGCCTAATAAAAAACATTTATTTTCATTGCAAG CTCGCTTTCTTGCTGTCCAATTTCTATTAAAGGTT CCTTTGTTCCCTAAGTCCAACTACTAAACTGGGG GATATTATGAAGGGCCTTGAGCATCTGGATTCTG CCTAATAAAAAACATTTATTTTCATTGCAA’ UTR HuR-BS TTTAATAGGGTGAGCTTGAGAGTTTTCTTTCTTT 57CTGTTTTTTTTTTTTTTGACTAATTTCACATGCTC TAA’ UTR HuR-BS-3x TTTAATAGGGTGAGCTTGAGAGTTTTCTTTCTTT 58CTGTTTTTTTTTTTTTTGACTAATTTCACATGCTC TAATTTAATAGGGTGAGCTTGAGAGTTTTCTTTC TTTGTGTTTTTTTTTTTTTTGACTAATTTCACATG CTCTAATTTAATAGGGTGAGCTTGAGAGTTTTCT TTCTTTCTGTTTTTTTTTTTTTTGACTAATTTCACA TGCTCTAA’ UTR CD47-3 ’termA GAGAGTTTTCTTTCTTTCTGTTTTTTTTTTTTGTTT 59TTTTTTTTTTTTTTTTTTTTTTTTTTTTTGACTAATT TCACATGCTCTAAAAACCTTCAAAGGTGATTATT TTTCTCCTGGAAACTCCAGGTCCATTCTGTTTAA ATCCCTAAGAATGTCAGAATTAAAATAACAGGG CTATCCCGTAATTGGAAATATTTCTTTTTTCAGG ATGCTATAGTCAATTTAGTAAGTGACCACCAAAT TGTTATTTGCACTAACAAAGCTCAAAACACGAT AAGTTTACTCCTCCATCTCAGTAATAAAAATTAA GCTGTAATCAACCTTCTAGGTTTCTCTTGTCTTA AAATGGGTATTCAAAAATGGGGATCTGTGGTGT ATGTATGGAAACACATACTCCTTAATTTACCTGT TGTTGGAAACTGGAGAAATGATTGTCGGGCAAC CGTTTATTTTTTATTGTATTTTATTTGGTTGAGGG ATI'TTI'TIATAAACAGTI'TTACT'TGTG'TCATAT'TTTAAAATTACTAA’ UTR CD47-3’termB GCCATTTTTTTGCAGTGATTTGAAGACCAAAGTT 60GTTTTACAGCTGTGTTACCGTTAAAGGTTTTTTTT TTTATATGTATTAAATCAATTTATCACTGTTTAAAGCTTTGAATATCTGCAATCTTTGCCAAGGTACTAttorney Docket No. TORQ-012 / 02WO 339010-2071TTTTTATTTAAAAAAAAACATAACTTTGTAAATA TTACCCTGTAATATTATATATACTTAATAAAACA TTTTAAGCTATTTTGTTGGGCTATTTCTATTGCTG CTACAGCAGACCACAAGCACATTTCTGAAAAAT TTAATTTATTAATGTATTTTTAAGTTGCTTATATT CTAGGTAACAATGTAAAGAATGATTTAAAATAT TAATTATGAATTTTTTGAGTATAATACCCAATAA GCTTTTAATTAGAGCAGAGTTTTAATTAAAAGTT TTAAATCAGTC3’ UTR DEN2 AAAGCAAAACTAACATGAAACAAGGCTAGAAGT 61CAGGTCGGATTAAGCCATAGTACGGAAAAAACT ATGCTACCTGTGAGCCCCGTCCAAGGACGTTAA AAGAAGTCAGGCCATCATAAATGCCATAGCTTG A GT A A ACT ATGCAGCCTGT A GCTCCACCTGAGA AGGTGTAAAAAATCCGGGAGGCCACAAACCATG GAAGCTGTACGCATGGCGTAGTGGACTAGCGGT TAGAGGAGACCCCTCCCTTACAAATCGCAGCAA CAATGGGGGCCCAAGGCGAGATGAAGCTGTAGT CTCGCTGGA A GGA CT AG AGGTTA GA GG AG ACCC CCCCGAAACAAAAAACAGCATATTGACGCTGGG AAAGACCAGAGATCCTGCTGTCTCCTCAGCATC ATTCCAGGCACAGAACGCCAGAAAATGGAATGGTGCTGTTGAATCAACAGGTTCT

[0306] The PKD2 cassettes were driven by an hPGK promoter, as the hPGK promoter showed robust PKD2 mRNA expression across multiple experiments, and transcription was terminated by a bGHpA polyadenylation signal. As shown in Figs. 7B and 7C, all PKD2 cassettes expressed PKD2 mRNA and PC2 protein, respectively, above the mock-transfected control. A cassette containing a synthetic 5’ UTR (PTR285) yielded a 2.1 -fold increase in both PKD2 mRNA and PC2 protein levels relative to a cassette without UTRs (PTR101) (Figs. 7B-7C). A cassette containing a hACTB 5’ UTR (PTR300) produced the highest average levels of PC2 protein per mRNA (Fig. 7D). Finally, a positive relationship was observed between PKD2 mRNA and PC2 protein levels (Fig. 7E). Together these results demonstrate that PKD2 / PC2 expression can be modulated with the use of different UTRs, providing an arsenal of cassettes that can be utilized depending on desired expression.Example 6 — In Vivo Expression of AAV-Packaged PKD2 Cassettes in Mice

[0307] This Example aims to show that the PKD2 cassettes designed based on Example 1 above can be expressed in vivo after delivery’ by recombinant AAV.k20 viruses.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0308] As shown in Fig. 8A, AAV.k20-packaged PKD2 cassettes were intravenously administered to 6-week-old naive mice (C57B1 / 6) via tail vein injection at a dose of 1E14, 3E14, or 8.6E14 vg / kg body weight. The mice were necropsied 28 days post- treatment and kidneys were harvested for expression analysis. A description of the AAV.k20-PKD2 cassettes is provided in Table 6 below.Table 6.AAV ID PTRID Description SEQ ID NO AAV075 PTR099 5ITR145 -CB A-PKD2 Acpg-3XHA-bGHpA-3ITRl 45 157 AAV076 PTR087 5ITR145-CBA-PKD2coAcpg-3XHA-bGHpA-3ITRI45 66 AAV127 PTR095 5ITR145-hPGK-PKD2Acpg-3XHA-bGHpA-3ITR145 156 AAV128 PTR101 5ITR 145 -hPGK-PKD2co Acpg-3 XII A-bGHp A-3I TR 145 67

[0309] AAV vector genome copies and PKD2 mRNA expression in the kidney were assessed following treatment via dPCR and RT-dPCR, respectively. AAV.k20-packaged PKD2 cassettes containing either CBA or hPGK promoters driving expression of either a codon-optimized and CpG-depleted PKD2 transgene (AAV 076, AAV128) or CpG-depleted PKD2 transgene (AAV 075, AAV127) had similar vector genome biodistribution copies in the kidney at matched doses (Fig.8B). At the RNA level, AAV.k20-packaged PKD2 cassettes containing a CBA promoter (AAV075 and AAV076) exhibited higher PKD2 mRNA copy numbers compared to AAV.k20-packaged PKD2 cassettes with a hPGK promoter (AAV127 and AAV128) at matched doses (Fig. 8C), Notably, the PKD2 mRNA copy numbers from the CBA vectors were three- to four-fold higher than the corresponding hPGK vectors at the same dose (Fig. 8C).

[0310] PC2 protein expression was determined via immunohistochemistry staining of kidney sections with an anti-HA antibody and 3,3'-diaminobenzidine (DAB) as a chromogen. The percentage of total PC2-HA+ cells in the kidney was determined via HALO image analysis software by counting the number of DAPI cells and the number of DAPI+ / 'PC2-HA+ cells. All A / XV.k20-packaged PKD2 cassettes expressed PC2 protein in the mouse kidney (Figs. 8D-8E). The percentage of total cells that were PC2-HA + in the kidneys of treated mice ranged from 0.6 to 2.6% across all four vectors (AAV075, AAV076, AAV127, and AAV128) and each dose (1E14, 3E14, and 8.6E14) (Fig. 8E).Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0311] The transduction efficiency of key kidney cells types, including proximal tubules, distal tubules, and collecting duct cells, were determined via immunofluorescence in mice treated with AAV128 and AAV076 (Figs. 8F-8G). Proximal tubules were identified via OATl-positive staining, distal tubules were identified via ECAD-positive and OAT1 -negative staining and collecting duct cells were identified via AQP2 positive staining. Each transduced key cell type was identified via the staining described above in addition to localization with PC2-HA tag positivity. Representative images showing transduction of proximal tubule, distal tubule, and collective duct cells with AAV.k20-packaged PKD2 cassettes are shown in Fig. 8F (white arrows). Distal tubule cells were transduced the most ranging from 1.9-3.4%, followed by collecting duct cells ranging from 1.4-1,7%, and lastly proximal tubule cells with about 0.5% being PC2-HA+ (Fig. 8G).

[0312] Together, these data demonstrate that AAV,k20-packaged PKD2 cassettes containing a CBA or hPGK promoter driving expression of a codon-optimized and CpG-depleted PKD2 transgene can result in PC2 protein expression in mouse kidney following administration in vivo. These cassettes also demonstrated the capability to transduce key kidney cell types (those from which cysts arise in ADPKD) m vivo, including proximal tubule cells, distal tubule cells, and collecting duct cells.Example 7 — In Vivo Expression of AAV-Packaged PKD2 Cassettes in Pigs

[0313] This Example aims to show that the PKD2 cassettes designed based on Example 1 above can be expressed in vivo following retrograde ureteral administration (RUA) of recombinant AAV.k20 viruses,

[0314] AAV076 or AAV128, which are described m Table 6 of Example 6 above, were administered to 11-13-week-old naive pigs via unilateral retrograde ureteral administration (RETA) (Fig. 9A). The pigs were necropsied 28 days post-treatment, and the kidneys, liver, ureter, and bladder were collected for vector biodistribution and expression analysis.

[0315] AAV vector genome copies and PKD2 mRNA expression in harvested tissues were assessed via dPCR and RT-dPCR, respectively. Kidney expression was analyzed and represented by either whole kidney, or regionally by cortex, inner medulla, and outer medulla. Vector genome copies in the whole kidney ranged from 1.6e4 to 4.8e4 copies per 1000 ng gDNA (Fig. 9B) and PKD2 mRNA copies in the whole kidney ranged from 1.9e3 to 8.8e3 copies per 100 ng cDNA (Fig. 9C) in pigs treated with AAV076 or AAV128 at a dose of 3E14.Attorney Docket No. TORQ-012 / 02WO 339010-2071

[0316] PC2 protein expression was determined via immunohistochemistry staining of kidney sections with an anti -HA antibody and DAB as a chromogen. The majority of HA positivity was seen in the cranial and caudal poles of the kidney (Fig.9D) compared to formulation buffer control (Fig. 9E). The percentage of PC2-HA+ tissue area was determined from whole sections of the kidney using Image!. The percentage of tissue area that was PC2-HA+ in the pig kidney ranged from 0.9 to 3.5% at a dose of 3E14 vg / kidney (Fig. 9F). Cellular morphology confirmed transduction in proximal tubules, distal tubules, and collecting duct cells of the kidney of treated pigs (Fig. 9G)

[0317] An additional in vivo study in pigs was performed to evaluate AAV076 transduction at multiple doses. AAV076 is described in Table 6 of Example 6 above. A AV076 was administered to 11-13-week-old naive pigs via bilateral RUA at either 1E14, 3E14, or 1E15 vg / kidney. Pigs were sacrificed 28 days following RUA. AAV vector genome copies and the expression of PKD2 mRNA in the kidney, liver, heart, bladder, and ureter were assessed via dPCR and RT-dPCR, respectively. The kidneys were represented by either whole kidney or regionally by cortex, inner medulla, and outer medulla. The liver, heart, bladder, and ureter are represented as whole tissue. Vector genome copies in the whole kidney ranged from 1.9e4 to 2.2e5 copies per 1000 ng gDNA across all three AAV doses administered to pigs (Fig. 10A) PKD2 mRNA copies in the whole kidney ranged from 1.1 e4 to 5.3e4 copies per 100 ng cDNA across all three AAV doses administered to pigs (Fig. 10B). AAV vector genomes were detected in the liver from the 3E14 and 1E15 dose groups ranging from 9.2e3 to 1.9e6 copies per 1000 ng gDNA (Fig. 10C, left panel). PKD2 mRNA copies in the liver were only detectable m the 1E15 dose group and the average was 2.1e4 copies per 100 ng cDNA (Fig. 10C, right panel). AAV vector genomes were detected in the heart from the 3E14 and 1E15 dose groups ranging from 4.6e2 to 1.3e4 copies per 1000 ng gDNA (Fig. 10D, left panel). PKD2 mRNA copies in the heart were detectable in the 3E14 and 1E15 dose groups ranging from 1.1 e2 to 1.5e4 copies per 100 ng cDNA (Fig. 10D, right panel). AAV vector genomes were detected in the bladder from the 3E14 and 1E15 dose groups ranging from 7.4e2 to 2.2e3 copies per 1000 ng gDNA (Fig. 10E, left panel). PKD2 mRNA copies in the bladder were detectable at all three doses ranging from 5.0el to 1.4e2 copies per 100 ng cDNA (Fig. 10E, right panel). AAV vector genomes were detected in the ureter from all three dose groups ranging from 1.0e3 to 1.5e5 copies per 1000 ng gDNA (Fig. 10F, left panel). PKD2 mRNA copies in the ureternoAttorney Docket No. TORQ-012 / 02WO 339010-2071were detectable at the 3E14 and 1E15 dose groups ranging from 3.7e2 to 1.3e3 copies per 100 ng cDNA (Fig. 10F, right panel).

[0318] PC2 protein expression was determined via immunohistochemistry staining as described above. The majority of PC2-HA positivity was seen in the cranial and caudal poles of the kidney (Fig. 10G). The percentage of tissue area that was PC2-HA+ in the whole pig kidney ranged from 1.5 to 5.7% across all three AAV doses evaluated (Fig. 10H).

[0319] Together, these data demonstrate that AAV076 and AAV128 can successfully transduce the kidney and lead to robust PC2 protein expression. Importantly, all vectors at all tested doses were well tolerated with no definitive changes among body weights, organ weights, clinical observations, clinical pathology parameters, or mortality in pigs following RUA in vivo.Example 8 — Demonstration of Biological Activity from AAV076 in an ADPKD2 Disease Mouse Model

[0320] This Example aims to evaluate the biological effect of an AAV.k20-packaged PKD2 cassette in a mouse model of ADPKD2.

[0321] A conditional Pkd2 knockout mouse was used as an ADPKD2 disease model for evaluation of AAV076. The mouse contains three transgenic genes consisting of a Pkd2 ° '' transgene, a Pax8rtTAallele, and a TetOCrsallele. Following administration of doxycycline at 50 mg / kg via intraperitoneal injection, Cre recombinase expression is activated, resulting in renal epithelial cellspecific Pkd2 knockout, which leads to cyst formation in kidneys. In this Example, ADPKD2 mice were injected with AAV076, described in Table 6 above, at post-natal day 1 via temporal facial vein injection at 1E14 vg / kg body weight. PA-<72-knockout was induced via three consecutive intraperitoneal injections of doxycycline at post-natal day 11, 12, and 13. Mice were necropsied at post-natal day 28, terminal body weights and kidneys weights were measured, and kidneys were harvested for expression analysis. Fig. HA provides a schematic of the study design.

[0322] The kidney weight to body weight (KW / BW) ratio was the primary therapeutic efficacy readout for this study. The KW / BW ratio was calculated by taking the sum of both kidney weights and normalizing to the terminal body weight. Notably, PA 2-knockout mice treated with AAV076 resulted in a 25% reduction in KW / BW ratio compared to cystic control mice that received doxycycline but no AAV (+Dox) (Fig. 11B).

[0323] AAV vector genome copies and PKD2 mRNA expression in the kidney were assessed via dPCR and RT-dPCR, respectively. Vector genome copies in the kidneys of PAz / 2-knockout miceAttorney Docket No. TORQ-012 / 02WO 339010-2071were 6.6e3 copies per 1000 ng gDNA following administration of AAV076 (Fig. 11C). PKD2 mRNA expression in the kidneys of PAz / 2-knockout mice was 3.0e3 copies per 100 ng cDNA following administration of AAV076 (Fig. 11D).

[0324] Together, these data demonstrate that AAV076 can significantly improve the cystic phenotype in a mouse model of ADPKD2 following systemic delivery in vivo.

[0325] It will be appreciated by those skilled in the art that changes could be made to the embodiments described above without departing from the broad inventive concept thereof. It is understood, therefore, that this invention is not limited to the particular embodiments disclosed, but it is intended to cover modifications within the spirit and scope of the present invention as defined by the present description.INCORPORATION BY REFERENCE

[0326] All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entireties for all purposes, including International Application No. PCT / US2025 / 031815. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not be taken as an acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world.

Claims

Attorney Docket No. TORQ-012 / 02WO 339010-2071WHAT IS CLAIMED:

1. A nucleic acid molecule comprising a polynucleotide encoding a poly cystin-2 (PC2) protein, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 3-5 and 7-9.

2. A nucleic acid molecule comprising a polynucleotide encoding a polycystin-2 (PC2) protein, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 5 or SEQ ID NO: 9.

3. The nucleic acid molecule of claim 1 or 2, wherein the PC2 protein comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 1.

4. The nucleic acid molecule of any one of claims 1 -3, wherein the PC2 protein comprises an amino acid sequence of SEQ ID NO: 1.

5. The nucleic acid molecule of any one of claims 1-4, wherein the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 3-5 and 7-9.

6. The nucleic acid molecule of any one of claims 1-5, wherein the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 5 or SEQ ID NO: 9.

7. The nucleic acid molecule of any one of claims 1 -6, wherein the polynucleotide is operably linked to a promoter.

8. The nucleic acid molecule of claim 7, wherein the promoter is selected from the group consisting of: chicken beta actin (CBA) promoter, a human phosphoglycerate kinase (hPGK) promoter, a human glucose-6-phosphatase (hG6Pase) promoter, a mouse polycystic kidney disease 2 (mPKD2) promoter, a mouse E-cadherin promoter (mECAD), a mouse kidney-specific cadherinAttorney Docket No. TORQ-012 / 02WO 339010-2071(mKSPC) promoter, a mouse paired box gene 8 (mPax8) promoter, a human with-no-lysine kinase 1 promoter with a renal enhancer element (hWNKl-RP), a human phosphoglycerate kinase promoter with renal enhancer element (RP-hPGK), and a human truncated PKD2 promoter, a pig truncated PKD2 promoter, and a mouse truncated PKD2 promoter.

9. The nucleic acid molecule of claim 7 or 8, wherein the promoter is a CBA promoter.

10. The nucleic acid molecule of claim 7 or 8, wherein the promoter is an hPGK promoter.

11. The nucleic acid molecule of any one of claims 7-10, wherein the promoter comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 12-15 and 21-32.

12. The nucleic acid molecule of any one of claims 7-11, wherein the promoter comprises a nucleic acid sequence of any one of SEQ ID NOs: 12-15 and 21-32.

13. The nucleic acid molecule of any one of claims 7-9, 11, and 12, wherein the promoter comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 12.

14. The nucleic acid molecule of any one of claims 7-9, 11, 12, and 13 wherein the promoter comprises a nucleic acid sequence of SEQ ID NO: 12.

15. The nucleic acid molecule of any one of claims 7, 8, and 10-12, wherein the promoter comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 13.

16. The nucleic acid molecule of any one of claims 7, 8, 10-12, and 15, wherein the promoter comprises a nucleic acid sequence of SEQ ID NO: 13.Attorney Docket No. TORQ-012 / 02WO 339010-207117. The nucleic acid molecule of any one of claims 1-16, wherein the polynucleotide is operably linked to a polyadenylation signal.

18. The nucleic acid molecule of claim 17, wherein the polyadenylation signal is a bovine growth hormone poly A (bGHpA) signal.

19. The nucleic acid molecule of claim 17 or 18, wherein the polyadenylation signal comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 18.

20. The nucleic acid molecule of any one of claims 17-19, wherein the polyadenylation signal comprises a nucleic acid sequence of SEQ ID NO: 18.

21. The nucleic acid molecule of claim 17, wherein the polyadenylation signal is a rabbit P-globin polyadenylation (rBGHpA) signal.

22. The nucleic acid molecule of claim 17 or 21, wherein the polyadenylation signal comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 33.

23. The nucleic acid molecule of claim 17, 21, or 22, wherein the polyadenylation signal comprises a nucleic acid sequence of SEQ ID NO: 33.

24. The nucleic acid molecule of any one of claims 1-23, wherein the polynucleotide is operably linked to a 5’ untranslated region (UTR).

25. The nucleic acid molecule of claim 24, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs:Attorney Docket No. TORQ-012 / 02WO 339010-207126. The nucleic acid molecule of claim 24 or 25, wherein the 5’ UTR comprises a nucleic acid sequence of any one of SEQ ID NOs: 34-49.

27. The nucleic acid molecule of any one of claims 24-26, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 35.

28. The nucleic acid molecule of any one of claims 24-27, wherein the 5’ UTR comprises a nucleic acid sequence of SEQ ID NO: 35.

29. The nucleic acid molecule of any one of claims 1-28, wherein the polynucleotide is operably linked to a 3 ’ UTR.

30. The nucleic acid molecule of claim 29, wherein the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 50-61.

31. The nucleic acid molecule of claim 29 or 30, wherein the 3’ UTR comprises a nucleic acid sequence of any one of SEQ ID NOs: 50-61.

32. The nucleic acid molecule of any one of claims 29-31, wherein the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 58.

33. The nucleic acid molecule of any one of claims 29-32, wherein the 3’ UTR comprises a nucleic acid sequence of SEQ ID NO: 58.

34. The nucleic acid molecule of any one of claims 1-23, wherein the polynucleotide is operably linked to a 5’ UTR and a 3’ UTR.Attorney Docket No. TORQ-012 / 02WO 339010-207135. The nucleic acid molecule of claim 34, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any one of SEQ ID NOs: 34-49; and the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to any one of SEQ ID NOs: 50-61.

36. The nucleic acid molecule of claim 34 or 35, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 44; and the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 50.

37. The nucleic acid molecule of claim 34 or 35, wherein the 5’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 37; and the 3’ UTR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to SEQ ID NO: 61.

38. The nucleic acid molecule of any one of claims 1 -37, wherein the polynucleotide comprises a translation initiation sequence.

39. The nucleic acid molecule of claim 38, wherein the translation initiation sequence comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least aboutAttorney Docket No. TORQ-012 / 02WO 339010-207195%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 20.

40. The nucleic acid molecule of claim 38 or 39, wherein the translation initiation sequence comprises a nucleic acid sequence of SEQ ID NO: 20.

41. The nucleic acid molecule of any one of claims 1 -40, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 108-155.

42. The nucleic acid molecule of any one of claims 1 -41, wherein the polynucleotide comprises a nucleic acid sequence of any one of SEQ ID NOs: 108-155.

43. The nucleic acid molecule of any one of claims 1 -40, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 109 or SEQ ID NO: 153.

44. The nucleic acid molecule of any one of claims 1 -41, wherein the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 109 or SEQ ID NO: 153.

45. The nucleic acid molecule of any one of claims 1 -40, wherein the polynucleotide comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 110 or SEQ ID NO: 154.

46. The nucleic acid molecule of any one of claims 1 -41, wherein the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 110 or SEQ ID NO: 154.

47. A vector comprising the nucleic acid molecule of any one of claims 1-46.Attorney Docket No. TORQ-012 / 02WO 339010-207148. The vector of claim 47, wherein the vector is a viral vector.

49. The vector of claim 48, wherein the viral vector is a recombinant adeno-associated virus (rAAV) vector.

50. The vector of claim 49, wherein the rAAV vector comprises a 5’ inverted terminal repeat (ITR) and a 3’ ITR, wherein the 5’ ITR flanks at the 5’ end of the nucleic acid molecule and the 3’ HR flanks at the 3’ end of the nucleic acid molecule.

51. The vector of claim 50, wherein the 5’ ITR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 16.

52. The vector of claim 50 or 51, wherein the 5’ ITR comprises a nucleic acid sequence of SEQ ID NO: 16.

53. The vector of any one of claims 50-52, wherein the 3’ ITR comprises a nucleic acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 17.

54. The vector of any one of claims 50-53, wherein the 3’ ITR comprises a nucleic acid sequence of SEQ ID NO: 17.

55. The vector of any one of claims 47-54, wherein the rAAV vector is derived from AAV1, AAV2, AAV2i8, AAV3, AAV3-B, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh8R, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAV-DJ, AAV LK03, AAVKPI, AAV.cc47, AAV.cc84, AAVrh74, AAV44-9, or a variant thereof.

56. The vector of any one of claims 47-55, wherein the rAAV vector comprises a polynucleotide sequence that is at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to any one of SEQ ID NOs: 65-107 and 156-160.Attorney Docket No. TORQ-012 / 02WO 339010-207157. The vector of any one of claims 47-56, wherein the rAAV vector comprises a polynucleotide sequence of any one of SEQ ID NOs: 65-107 and 156-160.

58. The vector of any one of claims 47-57, wherein the rAAV vector comprises a polynucleotide sequence that is at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 66 or SEQ ID NO: 158.

59. The vector of any one of claims 47-58, wherein the rAAV vector comprises a polynucleotide sequence of SEQ ID NO: 66 or SEQ ID NO: 158.

60. The vector of any one of claims 47-57, wherein the rAAV vector comprises a polynucleotide sequence that is at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 67 or SEQ ID NO: 159.

61. The vector of any one of claims 47-57 and 60, wherein the rAAV vector comprises a polynucleotide sequence of SEQ ID NO: 67 or SEQ ID NO: 159.

62. An rAAV particle comprising:(a) the nucleic acid molecule of any one of claims 1-46, or the vector of any one of claims 47 to 61; and(b) a capsid of AAV1, AAV2, AAV2i8, AAV3, AAV3-B, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh8R, AAV9, AAV 10, AAVrhlO, AAV11, AAV 12, AAV13, AAV-DJ, AAVLK03, AAV KIN, AAV.cc47, AAV.cc84, AAVrh74, AAV44- 9, or a variant thereof.

63. The rAAV particle of claim 62, wherein the rAAV particle transduces kidney cells.

64. The rAAV particle of claim 62 or 63, wherein the rAAV particle transduces distal tubule cells, proximal tubule cells, and / or collecting duct cells.

65. The rAAV particle of any one of claims 62-64, wherein the capsid protein is an AAV9 capsid variant.Attorney Docket No. TORQ-012 / 02WO 339010-207166. The AAV particle of any one of claims 62-65, wherein the capsid protein is an AAVkl3 capsid protein.

67. The AAV particle of claim 66, wherein the AAV.kl 3 capsid protein comprises an amino acid sequence of SEQ ID NO: 63 with 1, 2, 3, 4, or more amino acid substitutions at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

68. The AAV particle of claim 66 or 67, wherein the AAV.kl 3 capsid protein comprises an amino acid sequence of SEQ ID NO: 63 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

69. The AAV particle of any one of claims 66-68, wherein the AAV.kl 3 capsid protein consists of an amino acid sequence of SEQ ID NO: 63 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

70. The AAV particle of any one of claims 66-69, wherein the AAV.kl 3 capsid increases transduction of kidney cells by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, or more, compared to a wild-type AAV9 capsid.

71. The AAV particle of any one of claims 62-65, wherein the capsid protein is an AAVk20 capsid protein.

72. The AAV particle of claim 71, wherein the AAV.k20 capsid protein comprises an amino acid sequence of SEQ ID NO: 64 with 1, 2, 3, 4, or more amino acid substitutions at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

73. The AAV particle of claim 71 or 72, wherein the AAV.k20 capsid protein comprises an amino acid sequence of SEQ ID NO: 64 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.Attorney Docket No. TORQ-012 / 02WO 339010-207174. The AAV particle of any one of claims 71-73, wherein the AAV.k20 capsid protein consists of an amino acid sequence of SEQ ID NO: 64 at positions 452-458, wherein positions 452-458 of the AAV capsid protein are numbered with reference to SEQ ID NO: 62.

75. The AAV particle of any one of claims 71-74, wherein the AAV.k20 capsid increases transduction of kidney cells by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, or more, compared to a wild-type AAV9 capsid.

76. A pharmaceutical composition, comprising the nucleic acid molecule of any one of claims 1-46, the vector of any one of claims 47-61, or the rAAV particle of any one of claims 62-75, and a pharmaceutically acceptable excipient.

77. A method of treating a disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of the nucleic acid molecule of any one of claims 1-46, the vector of any one of claims 47-61, the rAAV particle of any one of claims 62-75, or the pharmaceutical composition of claim 76.

78. The method of claim 77, wherein the nucleic acid molecule of any one of claims 1-46, the vector of any one of claims 47-61, the rAAV particle of any one of claims 62-75, or the pharmaceutical composition of claim 76 is administered via retrograde ureteral infusion.

79. The method of claim 77, wherein the nucleic acid molecule of any one of claims 1-46, the vector of any one of claims 47-61, the rAAV particle of any one of claims 62-75, or the pharmaceutical composition of claim 76 is administered via the arterial route.

80. The method of any one of claims 77-79, wherein the nucleic acid molecule of any one of claims 1-46, the vector of any one of claims 47-61, the rAAV particle of any one of claims 62-75, or the pharmaceutical composition of claim 76 is administered at a dose of about 1E11 vector genomes per kidney (vg / kidney) to about IE 16 vg / kidney.Attorney Docket No. TORQ-012 / 02WO 339010-207181. The method of any one of claims 77-79, wherein the nucleic acid molecule of any one of claims 1-46, the vector of any one of claims 47-61, the rAAV particle of any one of claims 62-75, or the pharmaceutical composition of claim 76 is administered at a dose of about 1E11 vector genomes per kilogram (vg / kg) body weight to about IE 15 vg / kg body weight.

82. The method of any one of claims 77-81, wherein the disease or disorder is autosomal dominant polycystic kidney disease (ADPKD).

83. The method of any one of claims 77-82, wherein the disease or disorder is ADPKD type 2 (ADPKD2).

84. The method of any one of claims 77-83, wherein the method inhibits, ameliorates, or reverses renal cyst development.

85. The method of any one of claims 77-84, wherein the method inhibits, ameliorates, or reverses decline in kidney function.

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