Gene therapy

Abstract

The present invention relates to a method of treating an ocular disorder comprising a step of intravitreal administration of a vector comprising a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor.

Description

[0001] GENE THERAPY

[0002] Cross reference to related applications

[0003] This application claims priority from US63 / 782,768 filed on 3 April 2025, the contents of which are hereby incorporated by reference.

[0004] Field of the Invention

[0005] The present invention relates to a vector for use in a method of treating an ocular disorder, wherein the vector comprises a transgene encoding a C5 inhibitor and wherein the method comprises a step of intravitreal administration of the vector. The vector may be used to treat a complement-mediated disorder, such as age-related macular degeneration (AMD).

[0006] Background of the Invention

[0007] Complement component 5 (C5) is a protein involved in the complement system. The complement system is a part of the innate immune system, and forms a first line of defence against infections by triggering inflammatory responses. The complement system comprises a number of proteins that are synthesised primarily by the liver and circulate in the blood as inactive precursors. The stimulation of the complement system, for instance by a foreign entity, triggers proteases in the system to cleave the precursors and release molecules such as cytokines to initiate a further amplifying cascade. The activation of the complement system can stimulate phagocytes, stimulate inflammation and activate the cell-killing membrane attack complex. The complement system can be activated by the classical pathway, lectin pathway and alternative pathway. All three pathways converge at the cleavage and activation of C3 by C3-convertase. This promotes cleavage of C5 into C5a and C5b by C5 convertase. The membrane attack complex (MAC), also known as the terminal complement complex (TCC) can then be formed from C5b binding sequentially to C6, C7, C8 and multiple C9 molecules, leading to cell lysis. Overactivation of the complement pathway can therefore trigger excessive cell death in many tissues, leading to a range of complement-mediated disorders.

[0008] The complement system plays a role in many diseases with an immune component, such as lupus nephritis and C3 glomerulopathy in the kidney, and dry age-related macular degeneration (AMD) in the eye. Dysregulation of these pathways therefore has clinical implications. For example, if the pathway is over stimulated or inappropriately stimulated, the balance of the complement system can be then disrupted, leading to a build-up of inflammatory factors e.g. in the kidney, which can lead to kidney pathology. Thus, it can be desirable to rebalance the cycles of the complement pathways, particularly by inhibiting C5 cleavage, and thereby reduce inflammation and excessive cell death. Thus, it would be therapeutically advantageous to inhibit C5 cleavage.

[0009] Gene therapy treatments, which are licensed for a number of indications including ocular indications and under development for many more, generally involve administration of a gene therapy vector comprising a transgene (i.e. a polynucleotide that encodes a therapeutic protein). Suitable gene therapy vectors include viruses, like adeno-associated virus (AAV), that have had most or substantially all of their native genetic material removed but retain the ability to transduce cells. For example, a recombinant AAV particle made up of a capsid encapsulating a recombinant genome comprising a transgene could be used. As a genetherapy may result in durable in vivo expression of a therapeutic gene of interest, it offers a potential advantage over conventional therapeutics which require repeated administration. Thus, there is a need for a means to treat complement-mediated disorders, including ocular disorders such as AMD, with an effective gene therapy.

[0010] Ocular diseases, like dry AMD, can in principle be treated using gene therapy targeted to the retina, for example by subretinal administration. However, successful subretinal administration requires a high level of technical skill, and must be performed in an operating theatre by a trained surgeon.

[0011] Intravitreal administration of a gene therapy vector is another option, and this can be performed as an outpatient procedure by a medical professional without surgical training and so is significantly more accessible. However, intravitreal gene therapy requires use of a vector which mediates high transduction and gene expression levels so that a therapeutically meaningful amount of therapeutic protein reaches the retina at a vector dose that is below the level at which adverse inflammatory responses may occur.

[0012] Thus, there is a need for a means to treat ocular disorders using an accessible gene therapy requiring minimal technical skill and equipment, which is sufficiently potent but which does not induce a clinically intolerable immune reaction (e.g. inflammation).

[0013] Summary of the Invention

[0014] The present invention is based on the surprising finding that C5 inhibitor peptides can be expressed in human cells and secreted while retaining complement inhibitory activity. C5 inhibitors are attractive peptides for use in gene therapy, for example using an AAV viral particle as a vector, for several reasons. Firstly, the small size of C5 inhibitors means that an encoding transgene can comfortably be accommodated within a recombinant AAV genome no larger than the native AAV genome. Secondly, the small size of such a protein means that it can be expressed in one cell type and, once secreted, diffuse to a target site where it can mediate its effect. For example, a gene therapy encoding a C5 inhibitor may transduce, and cause the transgene to be expressed in, certain ocular cells. The C5 inhibitor will then be secreted from these cells, allowing the inhibitor to come into contact with the extracellular surface of retinal cells. Additionally, the present inventors have surprisingly found that tick-derived C5 inhibitors, despite being derived from a non-human source, demonstrate a low immunogenicity in humans.

[0015] Furthermore, the inventors have determined that intravitreal gene therapy is particularly useful for ocular disorders the treatment of which does not depend on expression of the therapeutic protein within the cells directly affected by the disorder, i.e. the diseased cells. A gene therapy vector that has been administered intravitreally may transduce a variety of cell types or populations, causing them to produce the therapeutic protein. If the therapeutic protein is a secreted protein, the therapeutic effect is not reliant on whatever amount of therapeutic protein may be produced by transduced diseased cells; rather therapeutic protein produced and secreted by transduced, non-diseased cell types may contribute to the therapeutic effect on the diseased cells. In other words, healthy cell types within the retinamay act as "factories ’ to produce the therapeutic protein, which can then be secreted to affect the target diseased cells.

[0016] Transfected cells may secrete the therapeutic protein into the retinal extracellular space, i.e. the space in the retina which is outside of the plasma membrane of neurons and glia, thereby increasing the concentration of the therapeutic protein in the retinal extracellular space.

[0017] Accordingly, in one aspect, the present invention provides a vector for use in a method of treating an ocular disorder, wherein the vector comprises a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor and wherein the method comprises a step of intravitreal administration of the vector.

[0018] In a second aspect, the present invention provides a method of treating an ocular disorder, comprising a step of intravitreal administration of a vector comprising a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor.

[0019] In a third aspect, the present invention provides a use of a vector in the manufacture of a medicament for use in a method of treating an ocular disorder, wherein the method comprises a step of intravitreal administration of the vector, wherein the vector comprises a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor.

[0020] Description of the Figures

[0021] Figure 1: In vitro inhibition of the TCC formation in human serum using supernatants from HEK293T cells transfected with plasmids expressing potential complement inhibitors. The inhibition of the TCC formation is reflected by the low absorbance values at 450 nm compared to supernatant from GFP -transfected (control) cells. Data from two independent experiments. Figure 2: CirpT4 expression levels (non-codon-optimised transgene) in HEK293T cells were comparable when using NATIVEsp or AHSGsp signal peptides. (A) Western Blot analysis using 6x-His tag antibody (Invitrogen Catalog # MA1-21315-HRP) at 1:1000 dilution. (B) Graph showing expression levels (ng / ml) of CirpT4 in the cleared supernatant using the His tag ELISA Detection Kit (Genscript Cat. No.: L00436). Neg Ctrl (Negative control): untransfected HEK293T cells. WPRE: Woodchuck hepatitis virus Posttranscriptional Regulatory Element. Figure 3 : CirpT4-Co01_WPRE with human AHSG signal peptide produced the highest secreted levels of CirpT4 following HEK293T cells transfection. (A) Western Blot analysis using CirpT4 polyclonal antibody (Beacon Tx T00710 Lot: U878P568G0-4 / GH2097) at 1:1000 dilution. (B) Relative protein expression of CirpT4 quantified using Image Lab software. Neg Ctrl (Negative control): untransfected HEK293T cells.

[0022] Figure 4: In vitro TCC formation inhibition assay demonstrates that supernatant from cells transfected with the construct CirpT4-Co01 with AHSGsp and WPRE had the strongest inhibition effect on the TCC formation. Untransfected HEK293T cells (Neg Ctrl) showed no inhibition. Supernatant of HEK293T cells transfected with plasmids carrying native or codon-optimized CirpT4 coding sequences was used. Hycult HK3012 AP complement activity ELISA Kit assay was used in this experiment.

[0023] Figure 5: In vitro Alternative Complement Pathway (AP) inhibition by CirpT4 following AAV transduction with a codon-optimized CirpT4 gene, with and without WPRE. Hycult HK3012 AP complement activity ELISA Kit assay was used in this experiment. Negative control: untransfected HEK293T cells.

[0024] Figure 6: In silico prediction to generate (A) HLA class I bound peptides using NetMHC4.0 server and (B) HLA class II bound peptides using NETMHCIIpan4.1 server.

[0025] Figure 7: B-cell reactivity to CirpT4, human Complement Factor H (CFH), and Hemagglutinin (Hl). Approximately 60% of B-cell populations from donor 1 and 28% of B-cell populations from donor 2 secrete IgG reactive to Influenza Hemagglutinin 1 protein, while a very low percentage of B-cell populations secrete IgG reactive to CirpT4 and CFH. In this assay, Hemagglutinin was used as positive control and CFH was used as negative control. Figure 8: IVIg from healthy donors was used in an ELISA assay to quantify the presence of antibodies specific for CirpT4, as well as for the respective positive and negative controls Hemagglutinin (shaded circles with grey border) and human CFH (shaded circles with black border).

[0026] Figure 9: Diagram showing the classical, lectin, and alternative complement pathways. All three pathways converge on activation of C3 convertase, which cleaves C3 into C3a and C3b. C3b activates C5 convertase, which cleaves C5 into C5a and C5b. C5b promotes formation of the Membrane Attack Complex (MAC), also known as the Terminal Complement Complex (TCC). Inhibition of C5 convertase is able to reduce the complement activity of all three complement pathways.

[0027] Figure 10: The transduction efficiency of two vectors (with and without additional poly A sequences) was assessed to ensure no decrease in CirpT4 expression was observed with the longer transgene cassette / vector genome. It was found that the longer vector genome did not negatively impact in vitro CirpT4 expression and even led to a modest increase in protein produced (A). Expanding the vector genome beyond half of the length of the wild type AAV genome (approximately 4.7 kb) by adding extra polyA sequences prevented dimerization from occurring. Although CB A-CirpT4-WPRE-bGH expressed well, due to the relatively small size of this vector genome (~2.3 kb ITR-ITR) dimerization was frequently observed when vector quality was assessed by alkaline gel electrophoresis. Expanding the vector genome to 2.8 kb by adding non-homologous polyA sequences - well beyond the 2.35 kb threshold (half of the length of the wild type AAV genome) - significantly reduced dimer formation (B). Overall, expanding the vector genome size by incorporating additional polyA elements significantly reduced genome dimerization without compromising expression.

[0028] Figure 11 : Graph shows fold-change in protein binding to CirpT4-conjugated beads in the serum (A) and vitreous (B). Figure 11(C) shows vitreous binders following subtraction of the blank (naive beads). Figure 11(D) shows the same data as Figure 11(C) with a loglO scale on the Y axis. Figure 11(E) shows the data for C5 only, following the removal of unreliable binding partners.Figure 12: Human serum samples from 100 US donors were screened for pre-existing anti-CirpT4 antibodies using a direct anti-CirpT4 antibody ELISA. Presence of anti-CirpT4 antibodies is detected by an increase in absorbance (OD 450nm). Positive control is antiserum from rabbits immunized with AAV-CirpT4 viral vector.

[0029] Figure 13: Representative plot of Luciferase activity in the supernatant of Rituximab tagged iLite® CD20(+) cells incubated in complement preserved serum with varying concentrations of CirpT4 (IC50 = 6.7 nM), commercially-available C5 inhibitor (IC50 = 2.5 nM) and anti-C5 antibody (6 nM). Across three independent experiments, the mean IC50 (± SD) values were 6.5 ± 0.5 nM for CirpT4, 5.2 ± 1.1 nM for the anti-C5 antibody, and 2.3 ± 0.4 nM for the commercially-available C5 inhibitor.

[0030] Figure 14: MAC puncta observed in: (A) ARPE-19 exposed to tBHP and complement preserved human serum; (B) iPSC-derived RPE oxidatively stressed with tBHP and exposed to complement preserved human serum. All images xlO captured on EVOSM5000 microscope. In ARPE 19 and iPSC derived RPE monolayers, serum dependent MAC puncta were readily detected, whereas no puncta were observed in monolayers treated with heat inactivated serum.

[0031] Figure 15: (A) Illustrative image of 594nm channel detecting Alexa Fluor 594 goat anti-mouse IgG2a (Invitrogen Cat. # A21135; 1:1,000) for C5b-9; (B) processed QC image from (A) demonstrating accurate automated puncta quantification; (C) Plotted data acquired from ARPE-19 monolayers; (D) Plotted data obtained from iPSC-derived RPE monolayers. Automated quantification showed a median MAC deposition of 117.5 puncta (IQR 97.75-138) in ARPE 19 cells and 78.5 puncta (IQR 73.25-114.3) in iPSC derived RPE. The addition of CirpT4 produced a clear, dose dependent reduction in MAC puncta across both cell types. Figure 16: Longitudinal intraocular pressure measurements over a 25-week period in cynomolgus macaques that received in each eye a single intravitreal injection of vehicle control or AAV-C5inhib_GLP at either 7.5 x 1O10(low dose), 2.5 x 1011(mid-dose), or 7.5 x 1011(high dose) vg / eye on Day 0 (n = 6 per treatment group until Week 12, n = 3 per treatment between Weeks 16 and 25). Means are plotted for each treatment group.

[0032] Figure 17: Longitudinal slit lamp scoring of vitreous cells and vitreous haze over a 25 week period in cynomolgus macaques that received in each eye a single intravitreal injection of vehicle control or AAV-C5inhib_GLP at either 7.5 x 1O10(low dose), 2.5 x 1011(mid-dose), or 7.5 x 1011(high dose) vector genome / eye on Day 0 (n = 6 per treatment group until Week 12, n = 3 per treatment between Weeks 16 and 25). Individual data for treated (OS) and untreated (OD) eyes are shown. The bold line indicates the timepoint when immunosuppression was discontinued. The color code of the scoring scale is depicted on the right. “X” = data not available: Week 6 and Week 7 measurements were additional unplanned assessments performed only for Animal #3003 (mid-dose group) and all groups were reduced to n =3 after Week 13 (necropsy n = 3).

[0033] Figure 18: Longitudinal slit-lamp scoring of aqueous cells and aqueous flare over a 25 week period in cynomolgus macaques that received in each eye a single intravitreal injection of vehicle control or AAV-C5inhib_GLP at either 7.5 x 1O10(low dose), 2.5 x 1011(mid-dose),or 7.5 x 1011(high dose) vector genome / eye on Day 0 (n = 6 per treatment group until Week 12, n = 3 per treatment between Weeks 16 and 25). Individual data for treated (OS) and untreated (OD) eyes are shown. The bold line indicates the timepoint when immunosuppression was discontinued. The color code of the scoring scale is depicted on the right. “X” = data not available: Week 6 and Week 7 measurements were additional unplanned assessments performed only for Animal #3003 (mid-dose group) and all groups were reduced to n = 3 after Week 13 (necropsy n=3).

[0034] Figure 19: Longitudinal slit lamp scoring of retinal sheathing over a 25 week period of in cynomolgus macaques that received in each eye a single intravitreal injection of vehicle control or AAV-C5inhib_GLP at either 7.5 x 1O10(low dose), 2.5 x 1011(mid-dose), or 7.5 x 1011(high dose) vector genome / eye on Day 0 (n = 6 per treatment group until Week 12, n = 3 per treatment between Weeks 16 and 25). Individual data for treated (OS) and untreated (OD) eyes are shown. “X” = data not available: Week 6 and Week 7 measurements were additional unplanned assessments performed only for Animal #3003 (mid-dose group) and all groups were reduced to n =3 after Week 13 (necropsy n = 3). The bold line indicates the timepoint when immunosuppression was discontinued (week 5).

[0035] Figure 20: AAV-C5inhib_GLP Longitudinal CirpT4 protein expression in vitreous humor (A) over a 25-week and in aqueous humor (B) over an 18-week period in cynomolgus macaques that received in each eye a single intravitreal injection of vehicle control or AAV-C5inhib_GLP at either 7.5 x 1O10(low dose), 2.5 x 1011(mid-dose), or 7.5 x 1011(high dose) vector genome / eye on Day 0. The dot-dash line shows the mean expression level of all animals in each dosage group.

[0036] Figure 21: Longitudinal complement activity in serum samples (measured by TCC assay) of Cynomolgus macaques that received in each eye a single intravitreal injection of vehicle control or AAV-C5inhib_GLP at either 7.5 x 1O10(low dose), 2.5 x 1011(mid-dose), or 7.5 x 1011(high dose) vector genome / eye on Day 0. In this assay, an inhibitory effect on complement activity would lead to a reduction in percentage of terminal complement complex (TCC). The dashed horizontal lines represent the accepted range of this assay (80 and 120%). All week 2 samples were tested together and all remaining samples were tested together in another, independent run.

[0037] Figure 22: Schematic Diagram of AAV-C5inhib_GLP. Abbreviations: C5 = complement component 5; CBA = cytomegalovirus enhancer + chicken P-actin promoter; ITR = (truncated) inverted terminal repeat; Poly As = 3 x polyadenylation signals; SP = signal peptide; WPRE = Woodchuck Hepatitis Virus Post-transcriptional Regulatory Element.

[0038] Detailed Description

[0039] General definitions

[0040] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by a person skilled in the art to which this invention belongs.

[0041] In general, the term “ comprising" is intended to mean including but not limited to. For example, the phrase “a polynucleotide comprising a transgene" should be interpreted to meanthat the polynucleotide has a transgene, but the polynucleotide may comprise further elements. In some embodiments of the invention, the word “comprising" may be replaced with the phrase “consisting of . The term “consisting of is intended to be limiting. For example, the phrase “a viral particle consisting of a capsid and a recombinant genome" should be interpreted to mean that the viral particle has a capsid and a recombinant genome and contains no further components.

[0042] In some embodiments of the invention, the word “comprising” may be replaced with the phrase “consisting essentially of’ . The term “consisting essentially of’ means that specific further components can be present, namely those not materially affecting the essential characteristics of the subject matter.

[0043] The singular forms “a”, “an”, and “the” include plural referents unless the context clear dictates otherwise. Thus, for example, reference to “a vector” includes one or more instances or versions of such vectors.

[0044] The terms “protein” and “polypeptide” are used interchangeably herein, and are intended to refer to a polymeric chain of amino acids of any length.

[0045] The term “fragment” as used herein refers to a contiguous portion of a reference sequence. For example, a fragment of a C5 inhibitor may refer to at least 50, at least 60, at least 70, or at least 80 contiguous amino acids of the C5 inhibitor.

[0046] The terms “nucleic acid molecule”, “polynucleotide” and “nucleotide sequence” are intended to refer to a polymeric chain of any length of nucleotides, including deoxyribonucleotides, ribonucleotides, or analogues thereof. For example, the nucleic acid molecule, polynucleotide or nucleotide sequence may comprise DNA (deoxyribonucleotides) or RNA (ribonucleotides). The nucleic acid molecule, polynucleotide or nucleotide sequence may consist of DNA. The nucleic acid molecule, polynucleotide or nucleotide sequence may be mRNA. Since the nucleic acid molecule, polynucleotide or nucleotide sequence may comprise RNA or DNA, all references to T (thymine) nucleotides may be replaced with U (uracil).

[0047] For the purpose of this invention, in order to determine the percent identity of two sequences (such as two polynucleotide or two polypeptide sequences), the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in a first sequence for optimal alignment with a second sequence). The nucleotide or amino acid residues at each position are then compared. When a position in the first sequence is occupied by the same nucleotide or amino acid as the corresponding position in the second sequence, then the nucleotides / amino acids are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity = number of identical positions / total number of positions in the reference sequence x 100).

[0048] Typically, the sequence comparison is carried out over the length of the reference sequence. For example, if the user wished to determine whether a given (“test”) sequence has at least 80% identity to SEQ ID NO: 1, SEQ ID NO: 1 would be the reference sequence. To assess whether a sequence has at least 80% identity to SEQ ID NO: 1 (an example of a reference sequence), the skilled person would carry out an alignment over the length of SEQ ID NO: 1,and identify how many positions in the test sequence were identical to those of SEQ ID NO: 1. If at least 80% of the positions are identical, the test sequence is at least 80% identical to SEQ ID NO: 1. If the sequence is shorter than SEQ ID NO: 1, the gaps or missing positions should be considered to be non-identical positions.

[0049] To assess whether a sequence is at least 80% identical to a fragment of 80 amino acids of SEQ ID NO: 1, the skilled person would align SEQ ID NO: 1 to the test sequence, and determine which contiguous 80 amino acids of SEQ ID NO: 1 best align to the test sequence. The skilled person would then determine the number of positions in the test sequence that are identical to the 80 amino acids of SEQ ID NO: 1 which best align to the test sequence, and calculate the percentage identity as indicated above.

[0050] The skilled person is aware of different computer programs that are available to perform an alignment between two sequences. An alignment between two sequences can be accomplished using a mathematical algorithm. For example, an alignment may be performed using the Needleman and Wunsch algorithm (Needleman and Wunsch, A general method applicable to the search for similarities in the amino acid sequence of two proteins, 1970, J Mol Biol.;48(3):443- 53) which aligns the sequences optimally over the entire length).

[0051] Sequences of substantially different lengths may alternatively be aligned using a local alignment algorithm (e.g. Smith and Waterman algorithm (Smith and Waterman, Overlapping genes and information theory, 1981, J Theor Biol. ;91 (2):379-80) or Altschul algorithm (Altschul SF et al., Gapped BLAST and PSLBLAST: a new generation of protein database search programs, 1997, Nucleic Acids Res.;25(17):3389-402,; Altschul SF et al., A structurebased method for protein sequence alignment, 2005, Bioinformatics. ;21 (8): 1451 -6). Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

[0052] Amino acids (or nucleotides) “ corresponding to" specified positions of a specified SEQ ID NO may be amino acids at the specified positions of the particular SEQ ID NO recited. For example, an amino acid substitution “at a position corresponding to position X of SEQ ID NO: 5" (e.g. position 1 or DI) may be a substitution at the Xth (e.g. the 1st) amino acid in a sequence. Alternatively, an amino acid (position) “ corresponding to ” position X of a specified SEQ ID NO may be an amino acid which is not the Xth amino acid, but rather the amino acid that best aligns to position X in the specified SEQ ID NO. It is within the capabilities of the person skilled in the art to determine which amino acids in an alternative amino acid sequence “ correspond to" the specified positions in the specified SEQ ID NO. For example, the person skilled in the art merely needs to perform a sequence alignment of the alternative amino acid sequence with the specified SEQ ID NO using a suitable alignment algorithm such as that of Needleman and Wunsch described above, and determine which region of the alternative amino acid sequence best aligns to the specified positions in the specified SEQ ID NO. For example, the skilled person is able to align the alternative amino acid sequence with SEQ ID NO: 5 and determine which amino acid best aligns, and therefore corresponds to, e.g. position X or 1 of SEQ ID NO: 5. Similarly, an amino acid substitution corresponds to DIE if it comprises an E at the position that best aligns to position DI of SEQ ID NO: 5.In some cases, a polypeptide or polynucleotide of the invention may be identical to a reference nucleotide, “but for” one or more substitutions or codons encoding one or more substitutions. As used herein, “identical to SEQ ID NO: X, but for an amino acid substitution” means that the polypeptide or polynucleotide has the same sequence as SEQ ID NO: X, other than the specified substitution(s). For example, a polypeptide that is identical to SEQ ID NO: 5, but for a DIE substitution has a sequence that is otherwise identical to SEQ ID NO: 5, except at position 1, where the polypeptide comprises a E in place of an D. In order to determine whether a polypeptide is identical to a reference sequence but for a specific substitution, the person skilled in the art merely needs to perform a sequence alignment of the amino acid sequence of the polypeptide with the specified SEQ ID NO using a suitable alignment algorithm such as that of Needleman and Wunsch described above, and determine whether the sequence is the same at all positions other than the specified position. For example, the skilled person is able to align the amino acid sequence of a polypeptide with SEQ ID NO: 5 and determine whether it is identical to SEQ ID NO: 5, but for an amino acid substitution at position 1.

[0053] A “conservative” amino acid substitution is a substitution of an amino acid for an amino acid with similar physical properties. Accordingly, such a substitution can be expected to have a smaller impact on the structure and function of a polypeptide compared with a nonconservative amino acid substitution. Examples of conservative amino acid substitutions for each amino acid are set out in the table below.

[0054] Original amino acid residue Conservative substitutions

[0055] Ser (S) Thr, Gly, Asn

[0056] Arg (R) His, Lys, Glu, Gin

[0057] Leu (L) He, Met, Phe, Vai, Tyr

[0058] Pro (P) Ala, Thr, Gly

[0059] Thr (T) Pro, Ser, Ala, Gly, His, Gin

[0060] Ala (A) Pro, Gly, Thr, Ser

[0061] Vai (V) Met, He, Tyr, Phe, Leu

[0062] Gly (G) Ala, Thr, Pro, Ser

[0063] He (I) Met, Leu, Phe, Vai, Tyr

[0064] Phe (F) Met, Tyr, He, Leu, Trp, Vai

[0065] Tyr (Y) Phe, Trp, Met, lie, Vai, Leu

[0066] Cys (C) Ser, Thr, Met

[0067] His (H) Gin, Arg, Lys, Glu, Thr

[0068] Gin (Q) Glu, His, Lys, Asn, Thr, Arg

[0069] Asn (N) Asp, Ser, Gin

[0070] Lys (K) Arg, Glu, Gin, His

[0071] Asp (D) Asn, Glu, Gin

[0072] Glu (E) Gin, Asp, Lys, Asn, His, Arg

[0073]

[0074] Met (M) lie, Leu, Phe, Vai

[0075] The terms “wild-type” and “native” are used interchangeably herein, and are intended to describe something which is naturally occurring. For example, a “wild-type signal peptide” is a signal peptide which occurs in nature.All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety.

[0076] Intravitreal administration

[0077] The vectors of the invention may be for intravitreal administration, and may be used or for use in methods of treatment comprising a step of intravitreal administration.

[0078] Intravitreal administration refers to administration to the vitreous humour of the eye. In some embodiments, intravitreal administration comprises administration to the vitreous humour using a needle. Optionally, the intravitreal administration is a form of administration that can be carried out by a person who is not a trained surgeon. Optionally, the intravitreal administration is a form of administration that can be carried out by a person who is not a doctor, such as a nurse or optometrist.

[0079] Without wishing to be to bound by theory, it is believed that intravitreal administration of the polynucleotide or viral particle of the invention results in expression of the transgene in ocular cells. In some embodiments, the ocular cells are retinal and / or para-retinal cells and / or anterior ocular cell types.

[0080] Vector

[0081] The present invention relates to vectors. Vectors typically comprise polynucleotides comprising a transgene along with other elements required for expression of the transgene in target cells, such as promoters and poly A sequences (collectively, a transgene cassette).

[0082] Vectors include non-viral particles, such as a plasmid or a nucleic acid minicircle. Plasmids or nucleic acid minicircles may be formulated as a liposome, lipid nanoparticle, cationic polymer, microvesicle or exosome. Vectors of the invention also include viral particles. Optionally, the viral particle is an AAV particle, an adenovirus particle, a lentivirus particle, a retrovirus particle, or a herpes simplex virus particle. In a preferred embodiment, the viral particle is an AAV particle.

[0083] Transgene

[0084] The vectors of the invention comprise a transgene. The term dransgene" refers to a nucleic acid sequence (typically encoding a protein) comprising a gene of interest (commonly a heterologous gene). The transgene is generally not an AAV-derived sequence, and may for example encode a tick protein or a protein from another organism. The term “a transgene" should be construed as comprising one or more transgenes.

[0085] In the present invention, the transgene encodes a C5 inhibitor. Optionally, the C5 inhibitor is a tick-derived C5 inhibitor.

[0086] Complement component 5 (C5)

[0087] C5 is a key component of the downstream (terminal) complement pathway. C5 therefore functions downstream of the classical pathway, lectin pathway, and alternative pathway. C5 is activated by C5 convertase which cleaves C5 into C5a and C5b. C5b triggers formation ofthe terminal complement complex (TCC), also known as the membrane attack complex (MAC), leading to cell lysis. Aberrant activation of the complement pathway may therefore lead to tissue damage and lead to disease states. For example, overactivation of the complement pathway in the eye may lead to age-related macular degeneration (AMD) as the cells of the macula are destroyed by overactivation of the complement pathway.

[0088] Inhibition of C5 reduces complement activity. For example, C5 may be inhibited by preventing conversion of C5 to C5b, by preventing cleavage of C5 by the C5 convertase.

[0089] C5 inhibitors

[0090] The transgenes of the present invention encode C5 inhibitors, which may be “tick-derived” C5 inhibitors. Ticks typically express complement inhibitors in their saliva, in order to suppress the host immune response to the tick during its bloodmeal. As described in more detail below, different tick species express several different families of C5 inhibitors, such as the CirpT family, RaCI family, and OmCI family. A C5 inhibitor is “tick-derived if a gene encoding the C5 inhibitor (or a C5 inhibitor at least 90% identical thereto) is present in the genome of one or more tick species, for example a species of the order Ixodida, or if the C5 inhibitor is derived from a C5 inhibitor encoded by a gene present in the genome of one or more tick species, for example a species of the order Ixodida. In some embodiments, the tick-derived C5 inhibitor is not or does not comprise OmCI or a variant thereof.

[0091] In some embodiments, the C5 inhibitor is not an antibody or a fragment or variant thereof. In some embodiments, the C5 inhibitor consists of 200 amino acids or fewer. In some embodiments, the C5 inhibitor is 150 amino acids or fewer in length. In some embodiments, C5 inhibitor is 200 or fewer, 150 or fewer, 140 or fewer, 130 or fewer, 120 or fewer, 110 or fewer, or 100 or fewer amino acids in length. In some embodiments, the C5 inhibitor is 100 or fewer amino acids in length. When determining the length of a C5 inhibitor, or when determining how many amino acids a C5 inhibitor consists of, any signal peptide portion should not be included. In some embodiments, the C5 inhibitor consists of between 50 and 200, between 50 and 150, between 50 and 140, between 50 and 130, between 50 and 120, between 50 and 110, or between 50 and 100 amino acids. In some embodiments, the C5 inhibitor consists of between 75 and 175 amino acids.

[0092] In some embodiments, the C5 inhibitor is a non-mammalian C5 inhibitor, i.e. the C5 inhibitor has been derived from a non-mammalian source.

[0093] In some embodiments, the C5 inhibitors bind to C5, and / or prevent cleavage of C5 to C5b by C5 convertase, for example by sterically hindering the docking of C5 with the convertase. Preventing cleavage of C5 to C5b by C5 convertase may be measured by detecting deposition of the terminal complement complex (TCC), for example using a complement activity assay as described below.

[0094] In some embodiments, the C5 inhibitor reduces TCC formation by at least 80%, at least 85%, at least 90%, or at least 95%.Complement activity assay

[0095] The final step in all three complement pathways (classical, lectin, and alternative) is the formation of the terminal complement complex (TCC), also known as membrane attack complex (MAC). The activity of inhibitors of the proteins in the complement pathways (such as C5 inhibitors) may therefore be measured using a complement activity assay comprising detecting formation of the TCC, for example using an ELISA assay as set out below.

[0096] To determine the specific C5 inhibitory activity (i.e. the activity of the C5 inhibitor at a known concentration) of a C5 inhibitor, the following complement activity assay can be used. A solution comprising a known concentration of the C5 inhibitor is incubated with human serum in wells coated with lipopolysaccharide (LPS). For example, the solution may be incubated for 1 hour. Following incubation, the wells are washed and levels of the TCC quantified using an antibody specific to the TCC. For example, an antibody specific for a C9 neo-epitope specific to the interface between C5b, C8, and C9 in the TCC may be used. An equivalent solution not comprising the C5 inhibitor (i.e. a solution which is identical to the test solution, but for the C5 inhibitor) can be used as a negative control. The % inhibition of TCC formation, i.e. the C5 inhibitory activity, can be determined by dividing the TCC levels of the samples by those of the negative control and multiplying by 100.

[0097] In some embodiments, the C5 inhibitor can reduce TCC formation by at least 70% of the level of reduction shown by a C5 inhibitor consisting of the amino acid sequence of SEQ ID NO: 5 in an equivalent complement activity assay. In some embodiments, the C5 inhibitor can reduce TCC formation by at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 100% of the level of reduction shown by a C5 inhibitor consisting of the amino acid sequence of SEQ ID NO: 5 in an equivalent complement activity assay.

[0098] For the purposes of comparing the specific C5 inhibitory activity of a test C5 inhibitor to a C5 inhibitor consisting of the amino acid sequence of SEQ ID NO: 5, an equivalent complement activity assay is identical to the assay used to determine the complement activity of the test polypeptide, i.e. all conditions are kept the same (e.g. the same known concentration of C5 inhibitor, incubation time) except that the C5 inhibitor is a C5 inhibitor consisting of the amino acid sequence of SEQ ID NO: 5. Optionally, the specific C5 inhibitory activity is normalised for the concentration of the C5 inhibitor.

[0099] To determine the C5 inhibitory activity (i.e. the activity of the C5 inhibitor taking into account both the specific complement activity of the C5 inhibitor and the expression level of the C5 inhibitor) of a vector comprising a polynucleotide comprising a transgene encoding a C5 inhibitor, the following complement activity assay can be used.

[0100] A vector comprising a polynucleotide comprising a promoter and a transgene (comprising a signal peptide-encoding region) encoding the C5-inhibitor whose activity is to be tested is expressed in human cells, for example in HEK293T cells. As the encoded C5-inhibitor comprises a signal peptide, it is secreted into the supernatant of the cell sample. The supernatant is collected and a serial dilution prepared using ELISA dilution buffer. Human serum is mixed with the same volume of each supernatant solution and transferred to wells coated with lipopolysaccharide (LPS). For example, the solution may be incubated for 1 hour. Following incubation, the wells are washed and levels of the TCC quantified usingantibody specific to the TCC. For example, an antibody specific for a C9 neo-epitope specific to the interface between C5b, C8, and C9 in the TCC may be used. Supernatant from untransfected cells can be used as a negative control. The % inhibition of TCC formation at a given level of supernatant dilution, i.e. the C5 inhibitory activity, can be determined by dividing the TCC levels of the samples by those of the negative control and multiplying by 100.

[0101] In some embodiments, the vector has at least 70% of the C5 inhibitory activity of an equivalent vector comprising a promoter comprising the nucleotide sequence which is identical to SEQ ID NO: 39 and a transgene comprising a nucleotide sequence which is identical to SEQ ID NO: 21 in an equivalent complement activity assay. In some embodiments, the vector has at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 100% of the C5 inhibitory activity of an equivalent vector comprising a promoter comprising the nucleotide sequence which is identical to SEQ ID NO: 39 and a transgene comprising a nucleotide sequence which is identical to SEQ ID NO: 21 in an equivalent complement activity assay.

[0102] For the purposes of comparing the C5 inhibitory activity of a test vector comprising a polynucleotide comprising a transgene encoding a C5 inhibitor to an equivalent vector comprising a promoter comprising the nucleotide sequence which is identical to SEQ ID NO: 39 and a transgene comprising a nucleotide sequence which is identical to SEQ ID NO: 21, an equivalent complement activity assay is identical to the assay used to determine the complement activity of the test vector, i.e. all conditions are kept the same (e.g. time between transfection and collection of supernatant, serial dilution, incubation time) except that the vector is a vector comprising a promoter comprising the nucleotide sequence which is identical to SEQ ID NO: 39 and a transgene comprising a nucleotide sequence which is identical to SEQ ID NO: 21.

[0103] A complement activity assay may be carried out using a Hycult Human Alternative Complement Pathway ELISA Kit HK3012. A complement activity assay may be carried out as described in Example 3.

[0104] CirpT family

[0105] The CirpT family includes polypeptides which are structurally and functionally related to CirpTl, which was initially derived from Rhipicephalus pulchellus. The family also includes CirpT2, derived from Dermacentor andersonii: CirpT3 derived from Rhipicephalus sanguineus,' and CirpT4, derived from Amblyomma americanum. CirpT C5 inhibitors can bind to the peripheral macro globulin domain 4 (C5 MG4 domain) and / or the peripheral macro globulin domain 5 (C5 MG5 domain) of C5, thereby preventing cleavage of C5 to C5b by C5 convertase. The CirpT family is described in more detail in Reichhardt MP et al., An inhibitor of complement C5 provides structural insights into activation, 2020, Proc Natl Acad Sci U S A. 117(l):362-370.

[0106] In some embodiments, the C5 inhibitor is a polypeptide from the CirpT family. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 6, 7, 84 or 85.In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of:

[0107] X1X2X3EX4GHX5YX6TX7NVTVEX8GACX9X10X11RNX12X13PX14GETKALX15X16PCVX17 X18TCYAAX19RX20VNX21TLCX22NX23GVX24X25GCX26X27X28WX29X30X31GX32YPX33CC PX34X35 VCX36X37X38X39X40X41X42 (SEQ ID NO : 84),

[0108] wherein Xi to X42 are selected from the amino acids listed in Table 1 or a conservative substitution thereof.

[0109] Xi D / E X9l / V X, / l / L X25 P / Q / E X; iQ / N X41 - / T / S

[0110] X / l / V X10 F / Y X1KA / S X?6 H / R X;l T / K X42 - / S

[0111] X; Q / R X11 E / L Xj, R / A / D X27 F / V X Q / H

[0112] Xi H / R X12 T / V X / .. E / K X?8 H / E X D / P

[0113] X'. S / T X13 l / L X / i A / S X29 R / T X; / G / T / A / S

[0114] Xr. L / R / V X14 N / D X,, R / P X30 N / P XiKT / A

[0115] X / R / K X15 H / N x / :F / l X31 D / V X D / S / P / T

[0116] XKN / D X16 D / S / N X / 1 D / E / A X32 V / E Xi.. - / A

[0117] Tab! e 1

[0118]

[0119] In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of:

[0120] X1X2X3EX4GHX5YX6X43X7NVTX44EX8GACX9X10X11RNX12X13PX14GETKA X15X16PCV X17X18TCYAX45X19RX20VNX21TLCX22NX23GVX24X25GCX26X27X28WX29X30X31GX32YPX 33CCPX34X35VCX36X37X38X39X40X41X42 (SEQ ID NO:85),

[0121] wherein Xi to X45 are selected from the amino acids listed in Table 2 or a conservative substitution thereof.

[0122] X, D / E X9l / V X, / l / L X25 P / Q / E X; iQ / N / E Xn - / T / S X / l / V X10 F / Y Xn; A / S X?6 H / R Xu T / K Xi / - / S X; Q / R X11 E / L Xi, R / A / D X27 F / V X;'. Q / H Xi ; T / K Xi H / R X12 T / V X / ,. E / K X?8 H / E / Q X / , D / P X11 l / V X'. S / T X13 l / L X / i A / S X?9R / T X; / G / T / A / S Xr> A / E Xr. L / R / V X14 N / D X / / R / P X30 N / P XiKT / A

[0123] X / R / K X15 H / N X / ; F / l X31 D / V X;, D / S / P / T / N

[0124] XKN / D X16 D / S / N X / 1 D / E / A X32 V / E / l Xi.. - / A / V

[0125]

[0126] fable 2In some embodiments, the C5 inhibitor is a polypeptide selected from the group consisting of a CirpTl polypeptide, a CirpT2 polypeptide, a CirpT3 polypeptide, and a CirpT4 polypeptide. In some embodiments, the C5 inhibitor is a CirpT4 polypeptide.

[0127] In some embodiments, the C5 inhibitor can bind to C5 with an affinity (Kd) of less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than InM. The affinity may be measured using any suitable method, for example surface plasmon resonance (SPR).

[0128] In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is:

[0129] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 50, at least 60, at least 70, or at least 80 amino acids of any one of SEQ ID NOs: 1-5;

[0130] (b) at least 95% identical to a fragment of at least 80 amino acids of any one of SEQ ID NOs: 1-5;

[0131] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 1-5; or

[0132] (d) identical to any one of SEQ ID NOs: 1-5.

[0133] In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to a fragment of at least 80 amino acids of SEQ ID NO: 5. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to SEQ ID NO: 5. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 98% identical to SEQ ID NO: 5. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is identical to SEQ ID NO: 5. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is identical to SEQ ID NO: 5, but for one or more conservative amino acid substitutions at one or more positions corresponding to positions 1, 2, 3, 5, 8, 12, 18, 22, 23, 28, 30, 37, 42, 43, 51, 54, 60, 67, 78, 83 and 88 of SEQ ID NO: 5. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence that is identical to SEQ ID NO: 5, but for one or more, two or more, three or more, four or more, or five or more conservative amino acid substitutions, optionally wherein the conservative amino acid substitutions correspond to one or more of DIE, I2V, Q3R, S8T, R12K, N18D, I22V, F23Y, L28I, D30N, H37N, I42L, A43S, E51K, A54S, F60I, R67H, Q78N, Q83H, and T88A.

[0134] In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 21 and 44 to 46. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 21 and 44 to 46. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is identical to any one of SEQ ID NOs: 21 and 44 to 46.

[0135] RaCI family

[0136] The RaCI family includes polypeptides which are structurally and functionally related to RaCIl, initially derived from R. appendiculatus. RaCI5 is a homolog from the Rappendiculatus transcriptome. RaCI2 is derived from Rhipicephalus microplus,' RaCI3 from Dermacentor andersoni RaCI4 from Hyalomma marginatum,' and RaCI6 and RaCI7 from D. andersoni. The polypeptides of the RaCI family can bind to the MG1, MG2, and / or C5d domains of C5, thereby preventing cleavage of C5 to C5b by C5 convertase. The RaCI family is described in detail in lore et al., Structural basis for therapeutic inhibition of complement C5., 2016, Nature Structural & Molecular Biology, 23.

[0137] In some embodiments, the C5 inhibitor is from the RaCI family. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of any one of SEQ ID NOs: 12, 86, or 87.

[0138] In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of:

[0139] X1CX2X3X4X5CX6RX7X8X9X1OX11GX12X13X14X15X16X17CPX18GCLCVX19X2OX21X22X23X24 X25X26X27X28X29GTCX30X31L (SEQ ID NO: 86),

[0140] wherein Xi to X31 are selected from the amino acids listed in Table 3 or a conservative substitution thereof.

[0141] Xi Q M X9D / N Xi- (1 K X25N / K / V / A / I / T S \ i: \ 1 X10 H / N / A Xi8 S P T Q (> X26Q / D / V / Y X ; 1 \ Xu L / R / A Xi 9 Y 1. 1 1 V X27 E / N / P Xi \ \ K X12 N / K / E / V XN. () K R S \ X28 A / L / V \< K T 1 X13 S / R / A / K Xzl \ G i: V X29 N / D

[0142] X.. R 11 r \ X14 V / H x- T P \ S X30 F / Y

[0143] \- R T K S X15 T / I X“ ? (> D S X31 E / L / A / Q

[0144]

[0145] X X 1 \ 1. 1 X16 E / D / S / T / A X2I Y S \ V

[0146] Table 3

[0147] In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of:

[0148] QCX1X2X3X4CX5RX6X7X8X9X1OGX11X12X13X14X15X16CPX17GCLCVX18X19X2OX21X22X23X 24X25X26X27X28GTCX29X30L (SEQ ID NO: 87),

[0149] wherein Xi to X30 are selected from the amino acids listed in Table 4 or a conservative substitution thereof.

[0150] Xi s \ 1: \ X9H / N / A Xi- S P T Q X25Q / D / V

[0151] 1: \ X10 L / R Xi8 Y 1. 1 V X26E / N / P X ; V \ Xu N / K / E Xl9 () K R X27 A / L

[0152] X K T X12 S / R / A XN. \ ( 1 1 X28 N / D

[0153] \c R I I i: X13 V / H X T P \ X29 F / Y

[0154] X.. R T K X14 T / I x- (> D S X30 E / L / A / Q \- 1 V 1. X15 E / D / S / T X“ ? Y S \

[0155]

[0156] X X D \ X16 G / K X \ K V 1

[0157] Table 4In some embodiments, the C5 inhibitor is a polypeptide selected from the group consisting of a RaCIl polypeptide, a RaCI2 polypeptide, a RaCI3 polypeptide, a RaCI4 polypeptide, a RaCI5 polypeptide, and a RaCI7 polypeptide.

[0158] In some embodiments, the C5 inhibitor can bind to C5 with an affinity (Kd) of less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM. Affinity may be measured using any suitable method, for example SPR. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is:

[0159] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 50, at least 60, at least 65, at least 70, or at least 75 amino acids of any one of SEQ ID NOs: 8- 11, 36, or 37;

[0160] (b) at least 95% identical to a fragment of at least 70 amino acids of any one of SEQ ID NOs: 8-11, 36, or 37;

[0161] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 8- 11, 36, or 37; and / or

[0162] (d) identical to any one of SEQ ID NOs: 8-11, 36, or 37.

[0163] In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to any one of SEQ ID NOs: 8-11, 36, or 37. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 98% identical to any one of SEQ ID NOs: 8-11, 36, or 37. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is identical to any one of SEQ ID NOs: 8-11, 36, or 37. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence that is identical to any one of SEQ ID NOs: 8-11, 36, or 37, but for one or more, two or more, three or more, four or more, or five or more conservative amino acid substitutions.

[0164] In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 47 to 52. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 47 to 52. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is identical to any one of SEQ ID NOs: 47 to 52.

[0165] OmCI

[0166] OmCI is a C5 inhibitor derived from Ornithodoros moubata. OmCI binds to C5 and displaces the C5 C345C domain, thereby inhibiting cleavage of C5 to C5b by C5 convertase. OmCI is also known as nomacopan.

[0167] In some embodiments, the C5 inhibitor can bind to C5 and displace the C5 C345C domain. In some embodiments, the C5 inhibitor is an OmCI polypeptide. In some embodiments, the C5 inhibitor can bind to C5 with an affinity (Kd) of less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, less than 1 nM, or less than 0.5 nM. Affinity may be measured using any suitable method, for example SPR.In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is:

[0168] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 70, at least 80, at least 90, at least 100, at least 110, at least 120, or at least 130 amino acids of SEQ IDNO: 38 or 95 to 101;

[0169] (b) at least 95% identical to a fragment of at least 130 amino acids of SEQ ID NO: 38 or 95 to 101;

[0170] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 38 or 95 to 101; and / or

[0171] (d) identical to SEQ ID NO: 38 or 95 to 101.

[0172] In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to SEQ ID NO: 38 or 95 to 101. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 98% identical to SEQ ID NO: 38 or 95 to 101. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is identical to SEQ IDNO: 38 or 95 to 101.

[0173] In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 53. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 53. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is identical to SEQ ID NO: 53. In some embodiments, the C5 inhibitor comprises a polypeptide comprising an amino acid sequence that is identical to SEQ ID NO: 53, but for one or more, two or more, three or more, four or more, or five or more conservative amino acid substitutions.

[0174] Codon optimised

[0175] In some embodiments, the polynucleotide and / or the transgene is codon-optimised.

[0176] Codon-optimisation can improve expression of the encoded product from the nucleotide sequence, for example a C5 inhibitor nucleotide sequence, in a particular cell type, tissue and / or in a particular organism. For example, if a nucleotide sequence is codon-optimised for expression in human cells, the nucleotide sequence may be modified to increase the number of codons that may be favoured (in the sense that such codons may correspond to tRNA species which are more abundant than other tRNA species specific for the same amino acid) in human cells. The skilled person would appreciate that codon-optimising a sequence may not entail changing every codon, not least because a “favoured codon" may already be present at some positions. For a given native coding sequence, due to degeneracy of the genetic code there are numerous permutations of the sequence obtainable by altering one or more codons (without changing the sequence of the encoded polypeptide) with the aim of improving expression of the gene product. The performance of a given variant coding sequence is unpredictable and whether such a sequence is considered codon-optimised is determined empirically.

[0177] Such codon-optimisation may be subject to other factors. For example, favoured codons optionally may not be introduced at positions where doing so introduces CG dinucleotides(CpGs) into the sequence; this will still be considered to be codon-optimisation. In an embodiment, a favoured codon that ends with a C nucleotide will not be included in the portion of the coding sequence that is codon-optimised, where the adjacent downstream codon in the sequence begins with a G. For example, codon GCC encodes alanine. Optionally, where GCC is a favoured codon, it should not be used for encoding alanine where the next codon in the sequence begins with a G, such as codon GAC (or alternatively, the next codon -where possible - could be selected to avoid a G at the first position).

[0178] In some embodiments, the transgene and / or the polynucleotide is considered codon optimised for expression if, when part of a plasmid or a vector such as an AAV vector, it expresses at a higher level in HEK293T cells compared to an equivalent native transgene and / or polynucleotide in an equivalent plasmid or vector. An “equivalent” native transgene and / or polynucleotide is a transgene and / or polynucleotide that is otherwise identical (and encodes an identical polypeptide), i.e. has an identical nucleotide sequence but for the codon optimisation (for example, if the skilled person wanted to determine if a CirpT4 transgene was codon optimised, a native CirpT4 sequence such as SEQ ID NO: 21 could be used, or if the CirpT4 is a fragment or variant, a sequence based on SEQ ID NO: 21 and with the same codon usage pattern). Similarly, an “equivalent” vector is a vector which is otherwise identical, i.e. in the case of an AAV vector comprises the same capsid and recombinant genome comprising ITRs and expression cassette(s) comprising the transgene and / or polynucleotide, but for the codon optimisation of the transgene and / or polynucleotide.

[0179] Expression of the polypeptide encoded by the transgene and / or polynucleotide can be measured using an ELISA.

[0180] In some embodiments, the polynucleotide and / or the transgene is codon optimised if, when part of a vector, it expresses at a level that is a factor of at least 1.5, or a factor of at least 2 better in HEK293T cells compared to an equivalent native transgene and / or polynucleotide in an equivalent vector.

[0181] The presence of CpGs in a gene therapy vector may have an adverse effect on expression of a therapeutic transgene (e.g. the durability of expression). This is because CpGs may be methylated, and their methylation may lead to gene silencing thereby reducing expression. Also, it is possible that high CpG content could trigger a TLR response, increasing the risk of an adverse immune response. The transgene or the polynucleotide may comprise a reduced number of CpGs relative to the corresponding native sequence. For example, in some embodiments, the C5 inhibitor-encoding nucleotide sequence (transgene) comprises a reduced number of CpGs compared to a corresponding portion of a reference nucleotide sequence, such as a native gene sequence. In the context of determining the number of CpGs, a “corresponding portion” of a reference nucleotide sequence is the portion of the reference sequence which aligns to the transgene. For example, a transgene of the invention may comprise a polynucleotide encoding a fragment of a C5 inhibitor. In this case, the transgene should be aligned to the reference nucleotide sequence, and the number of CpGs in the transgene should be compared with the number of CpGs in the portion of the reference nucleotide sequence which aligns with the transgene.

[0182] In some embodiments, the transgene and / or the polynucleotide comprises fewer than 20, fewer than 15, fewer than 10, or fewer than 5 CpGs. In a preferred embodiment, the transgene and / or the polynucleotide is CpG-free.In some embodiments, the polynucleotide comprises a nucleotide sequence which is:

[0183] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 175, at least 200, at least 225, or at least 250 nucleotides of any one of SEQ ID NOs: 22-26; (b) at least 95% identical to a fragment of at least 250 nucleotides of any one of SEQ ID NOs: 22-26;

[0184] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 22- 26; and / or

[0185] (d) identical to any one of SEQ ID NOs: 22-26.

[0186] In some embodiments, the polynucleotide comprises a nucleotide sequence which is:

[0187] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 175, at least 200, at least 225, or at least 250 nucleotides of any one of SEQ ID NOs: 22, 23, 25 or 26;

[0188] (b) at least 95% identical to a fragment of at least 250 nucleotides of any one of SEQ ID NOs: 22, 23, 25 or 26;

[0189] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 22, 23, 25 or 26; and / or

[0190] (d) identical to any one of SEQ ID NOs: 22, 23, 25 or 26.

[0191] In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 22. In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 22. In some embodiments, the polynucleotide comprises a nucleotide sequence which is identical to SEQ ID NO: 22.

[0192] In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 54 to 63. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 54 to 63. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is identical to any one of SEQ ID NOs: 54 to 63.

[0193] In some embodiments, the polynucleotide and / or the transgene promotes expression of the C5 inhibitor at a higher level than an equivalent polynucleotide and / or transgene that is not codon-optimised.

[0194] In some embodiments, the polynucleotide and / or the transgene promotes expression of the C5 inhibitor at a higher level than a polynucleotide and / or transgene comprising an equivalent native transgene, optionally the native transgene of SEQ ID NO: 21.

[0195] In some embodiments, the polynucleotide and / or the transgene promotes at least 70% or at least 90% of the level of expression of a polynucleotide and / or transgene comprising SEQ ID NO: 22.

[0196] In some embodiments, the expression is measured by transfecting a HEK293T cell sample with a plasmid or an AAV vector comprising the polynucleotide and / or the transgene,harvesting the supernatant of the cell sample and measuring the level of the C5 inhibitor, for example using an antibody specific for the C5 inhibitor.

[0197] Signal peptide

[0198] Typically, a C5 inhibitor is initially expressed as a precursor "immature" form of polypeptide comprising a signal peptide. The signal peptide is typically removed during post-translational processing such that the "mature" form of the C5 inhibitor lacks a signal peptide.

[0199] In some embodiments, a polypeptide of the invention comprises a signal peptide. In some embodiments, the C5 inhibitor comprises the inhibitor’s native signal peptide. In some embodiments, the C5 inhibitor comprises a heterologous signal peptide. In some embodiments, the signal peptide is a human protein-derived signal peptide, optionally an alpha-2 -HS-glycoprotein / fetuin-A (AHSG) signal peptide. Optionally, the polypeptide of the invention does not comprise a signal peptide, i.e. is a "mature" polypeptide.

[0200] In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence encoding a signal peptide. In some embodiments, the signal peptide comprises the inhibitor’s native signal peptide. In some embodiments, the signal peptide comprises a heterologous signal peptide. In some embodiments, the signal peptide is a human protein-derived signal peptide, optionally an alpha-2-HS-glycoprotein / fetuin-A (AHSG) signal peptide.

[0201] When a polynucleotide comprising a nucleotide encoding a signal peptide, for example SEQ ID NO: 15 or 20, is expressed in human cells, the resulting "mature" polypeptide is typically secreted by the human cells. Accordingly, if a polynucleotide comprising a nucleotide encoding a signal peptide is expressed in a HEK293T cell sample, the polypeptide encoded by the polynucleotide will typically be present in the supernatant of the HEK293T cell sample. When a polypeptide, for example a C5 inhibitor, is expressed in cells transduced with a polynucleotide or transgene encoding it, such as retinal and / or para-retinal and / or anterior ocular cells, it may be secreted by said cells into the vitreous humour, aqueous humour and / or retinal extracellular space - for example if it encodes a signal peptide. The signal peptide may be a heterologous signal peptide. The concentration of the secreted protein in the retinal extracellular space may therefore be increased such that the secreted polypeptide is able to reach cells other than those from which it is being produced and secreted. Such cells may be retinal and / or para-retinal including subretinal cells, for example the diseased cells directly impacted in dAMD. For example, a C5 inhibitor expressed in transduced cells (which may be at the posterior or anterior of the eye, or both) may be secreted into the vitreous and / or aqueous compartments of the eye, leading to an increased concentration in the retinal extracellular space. Consequently, the C5 inhibitor may then act extrinsically, for example at the cell surface, with respect to cells of the retina and macula to downregulate pro-inflammatory complement activity locally (in addition to any intrinsic action of the C5 inhibitor in diseased cells which have been transduced and are able to express the C5 inhibitor from the transgene), thereby treating or ameliorating the pathology or symptoms of dry AMD. Accordingly, a polynucleotide encoding a C5 inhibitor typically comprises a nucleotide sequence encoding a signal peptide.In some embodiments, the polynucleotide comprises a nucleotide sequence which is:

[0202] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 200, at least 225, at least 250, at least 275, or at least 300 nucleotides of any one of SEQ ID NOs: 14-19;

[0203] (b) at least 95% identical to a fragment of at least 300 nucleotides of any one of SEQ ID NOs: 14-19;

[0204] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 14- 19; and / or

[0205] (d) identical to any one of SEQ ID NOs: 14-19.

[0206] In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 95% identical to a fragment of at least 300 nucleotides of SEQ ID NO: 15. In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 15. In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 15. In some embodiments, the polynucleotide comprises a nucleotide sequence which is identical to SEQ ID NO: 15.

[0207] In some embodiments, the polynucleotide comprises a nucleotide sequence which is:

[0208] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 200, at least 225, at least 250, at least 275, or at least 300 nucleotides of SEQ ID NO: 20; (b) at least 95% identical to a fragment of at least 300 nucleotides of SEQ ID NO: 20; (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 20; and / or (d) identical to SEQ ID NO: 20.

[0209] In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 95% identical to a fragment of at least 300 nucleotides of SEQ ID NO: 20. In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 20. In some embodiments, the polynucleotide comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 20. In some embodiments, the polynucleotide comprises a nucleotide sequence which is identical to SEQ ID NO: 20.

[0210] In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 64 to 66 or 70 to 76. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 64 to 66 or 70 to 76. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is identical to any one of SEQ ID NOs: 64 to 66 or 70 to 76.

[0211] In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 67 to 69 or 77 to 83. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 67 to 69 or 77 to 83. In some embodiments, the polynucleotide and / or the transgene comprises a nucleotide sequence which is identical to any one of SEQ ID NOs: 67 to 69 or 77 to 83.Immunogenicity

[0212] It is advantageous if the C5 inhibitor does not provoke a large immune response in a subject, as this improves the safety of a therapy comprising the C5 inhibitor, i.e. when the C5 inhibitor is administered to the subject, no more than a limited immune response is elicited. The immune response may be measured by contacting peripheral blood mononuclear cells (PBMCs) from a healthy donor with the C5 inhibitor and measuring the levels of INF -y secreted. Alternatively, the immune response may be measured by contacting B-cells derived from peripheral blood mononuclear cells (PBMCs) from different donors with the C5 inhibitor and detecting IgG secretion.

[0213] In some embodiments, the C5 inhibitor promotes a limited or negligible immune response in a subject. In some embodiments, the C5 inhibitor promotes a similar or smaller immune response to that elicited by an otherwise equivalent administration of human complement factor H (CFH) protein. In some embodiments, the C5 inhibitor promotes a smaller immune response than influenza haemagglutinin 1 (Hl), for example the Hl protein of SEQ ID NO: 102.

[0214] In some embodiments, the C5 inhibitor results in less IFN-y secretion than Hl, optionally wherein the C5 inhibitor results in less than 50% IFN-y secretion from PBMCs than that caused by Hl in an otherwise equivalent assay.

[0215] In some embodiments, fewer than 10% of the B-cells release IgGs in response to the C5 inhibitor.

[0216] Immunogenicity can also be predicted in silico, for example using the NetMHC4.0 server (Andreatta M, Nielsen M, Gapped sequence alignment using artificial neural networks: application to the MHC class I system, 2016, Bioinformatics;32(4):511-7 and Nielsen et al., Reliable prediction of T-cell epitopes using neural networks with novel sequence representations, 2003, Protein Sci., 12:1007-17) or the NETMHCIIpan4 server (Reynis son et al., Improved prediction of MHC II antigen presentation through integration and motif deconvolution of mass spectrometry MHC eluted ligand data, 2020, J Proteome Res, 19, 6, 2304-2315), which predict weak and strong binders from HLA class I and II peptides respectively. These servers can be used to predict the number of strong and weak binders against short peptide sequences derived from a polypeptide of interest. In some embodiments, the C5 inhibitor is predicted to give rise to fewer strong and / or weak binders than Hl.

[0217] Viral particle

[0218] In some embodiments, the vector is a viral particle, for example an AAV particle, an adenovirus particle, a lentivirus particle, or a herpes simplex virus particle.

[0219] The term “viral particle" relates to a typically replication-defective virus particle comprising (i) at least a portion of a viral genome (ii) a capsid and optionally (but not in the case of AAV), (iii) a lipidic envelope surrounding the capsid. The term “viral particle" includes recombinant adeno-associated viral (AAV) particles, comprising an AAV capsid and a genome comprising sequences designed to function in an AAV (such as ITR sequences).The term “viral genome” refers to the nucleic acid part of the viral particle disclosed herein, which may be packaged in a capsid.

[0220] In the present invention, the viral particle may be a recombinant adeno-associated viral particle (rAAV). The terms rAAV and AAV are used interchangeably herein, unless the context indicates otherwise.

[0221] The genomic organization of all known AAV serotypes is very similar. The genome of native (non-recombinant) AAV is a linear, single-stranded DNA molecule that is less than about 5,000 nucleotides in length. Inverted terminal repeats (ITRs) flank the unique coding nucleotide sequences for the non- structural replication (Rep) proteins and the structural (VP) proteins. The VP proteins (VP1, -2 and -3) form the capsid. The terminal 145 nucleotides, the ITRs, are self-complementary and are organized so that an energetically stable intramolecular duplex forming a T-shaped hairpin may be formed. These hairpin structures function as an origin for viral DNA replication, serving as primers for the cellular DNA polymerase complex. Following wild type (wt) AAV infection in mammalian cells the Rep genes (i.e. encoding Rep78 and Rep52 proteins) are expressed from the P5 promoter and the P19 promoter, respectively, and both Rep proteins have a function in the replication of the vector genome. A splicing event in the Rep ORF results in the expression of four Rep proteins (i.e. Rep78, Rep68, Rep52 and Rep40). However, it has been shown that the unspliced mRNA, encoding Rep78 and Rep52 proteins, in mammalian cells are sufficient for AAV vector production. Also in insect cells the Rep78 and Rep52 proteins suffice for AAV vector production. An AAV vector for use in gene therapy will be replication deficient, for example it may not comprise the genes encoding one or more of the four Rep proteins. The vector of the invention preferably does not comprise a gene encoding a Rep protein.

[0222] In some embodiments, the vector of the present invention is a viral particle. In some embodiments, the viral particle is an AAV, adenoviral, or lentiviral particle. In a preferred embodiment, the viral particle is an AAV particle.

[0223] In some embodiments, the viral particle is capable of delivering the transgene to ocular cells. In some embodiments the ocular cells are retinal and / or para-retinal cells, optionally including one or more cell types selected from retinal ganglion cells, amacrine cells, horizontal cells, bipolar cells, photoreceptor cells, cone cells, rod cells, Muller glial cells, retinal pigmented epithelium (RPE) cells and / or choroid cells. In some embodiments, the ocular cells are cells in the retinal pigment epithelial layer or cells in the choroidal capillary layer. In some embodiments the ocular cells are anterior ocular cell types such as cells of the ciliary body and / or iris, optionally in addition to the aforementioned retinal and para-retinal cells.

[0224] In some embodiments, the viral particle is capable of delivering the transgene to cells of the kidney, such as the podocytes.

[0225] A A V capsid

[0226] In some embodiments, the viral particle comprises a capsid, such as an AAV capsid.

[0227] AAV capsids are generally formed from three proteins, VP1, VP2 and VP3. The amino acid sequence of VP1 comprises the sequence of VP2. The amino acid sequence of VP2comprises the sequence of VP3. Differences among the capsid protein sequences of the various native and engineered AAV serotypes result in the use of different cell surface receptors for cell entry. In combination with alternative intracellular processing pathways, this gives rise to variation in tissue tropisms between the AAV serotypes.

[0228] In some embodiments, the capsid is an AAV2 capsid, an AAV2-derived capsid, an AAV5 capsid, an AAV5-derived capsid, an AAV6 capsid, or an AAV6-derived capsid.

[0229] Capsids derived from the capsid of a particular serotype, e.g. AAV2-derived capsids, may include an AAV capsid protein with an insertion, for example an insertion of the 7m8 amino acid sequence, an R100 insertion, or an LSV1 sequence replacement. The LSV1 replacement sequence and the 7m8 insertion sequence are known in the art (see, for example, U.S. Patent 9,193,956; U.S. Patent 9,233,133; U.S. Pub. No. US2021 / 0040501; and PCT / US2020 / 029895), as is the R100 insertion (PCT / US2017 / 032542).

[0230] An example sequence of an AAV2 capsid is given in SEQ ID NO: 35. SEQ ID NO: 35 represents the amino acid sequence of the AAV2 VP1 protein, and comprises within it the amino acid sequences of the AAV2 VP2 and VP3 proteins. For the purposes of the present invention, the AAV particle will be considered to comprise an AAV2 capsid if it comprises at least an AAV2 VP3, VP2, or VP1 protein. Optionally, the AAV particle will be considered to comprise an AAV2 capsid if it comprises at least an AAV2 VP1 protein. Optionally, the AAV particle will be considered to comprise an AAV2 capsid if it comprises an AAV2 VP1 protein, an AAV2 VP2 protein and an AAV2 VP3 protein.

[0231] An exemplary sequence of an AAV6 capsid is given in SEQ ID NO: 31. SEQ ID NO: 31 represents the amino acid sequence of the AAV6 VP1 protein, and comprises within it the amino acid sequences of the AAV6 VP2 and VP3 proteins. For the purposes of the present invention, the AAV particle will be considered to comprise an AAV6 capsid if it comprises at least an AAV6 VP3, VP2 or VP1 protein. Optionally, the AAV particle will be considered to comprise an AAV6 capsid if it comprises at least an AAV6 VP1 protein. Optionally, the AAV particle will be considered to comprise an AAV6 capsid if it comprises an AAV6 VP1 protein, an AAV6 VP2 protein and an AAV6 VP3 protein.

[0232] An AAV2-derived capsid is (i) a capsid comprising at least a fragment of a wild-type AAV2 capsid, for example an AAV2-derived capsid may comprise at least 100, at least 200, at least 300, at least 400, at least 500, at least 600 or at least 700 amino acids of a wild-type AAV2 capsid or (ii) a capsid having a sequence which has 100% identity to wild type AAV2 except for one or a small number of amino acid substitutions, and / or short insertions or replacements of up to about 15 amino acids in length. An AAV2-derived capsid may further comprise one or more substitutions compared to a wild-type AAV2 capsid. An AAV2-derived capsid may further comprise a fragment from the capsid of another AAV serotype. In some embodiments, the AAV2-derived capsid is any one of SEQ ID NO: 27-30.

[0233] In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of any one of SEQ ID NO: 27-30. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of anyone of SEQ ID NO: 27-30. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of any one of SEQ ID NO: 27-30. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to any one of SEQ ID NO: 27-30. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to any one of SEQ ID NO: 27-30. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence of any one of SEQ ID NO: 27-30.

[0234] In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of SEQ ID NO: 90. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of SEQ ID NO: 90. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of SEQ ID NO: 90. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO: 90. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to SEQ ID NO: 91. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 91.

[0235] An AAV5-derived capsid is (i) a capsid comprising at least a fragment of a wild-type AAV5 capsid, for example an AAV5-derived capsid may comprise at least 100, at least 200, at least 300, at least 400, at least 500, at least 600 or at least 700 amino acids of a wild-type AAV5 capsid, for example the AAV5 capsid of SEQ ID NO: 91 or (ii) a capsid having a sequence which has 100% identity to wild type AAV5 except for one or a small number of amino acid substitutions, and / or short insertions or replacements of up to about 15 amino acids in length. A capsid may further comprise one or more substitutions compared to a wild-type AAV5 capsid. A capsid may further comprise a fragment from the capsid of another AAV serotype. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 92.

[0236] In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of SEQ ID NO: 92. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of SEQ ID NO: 92. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of SEQ ID NO: 92. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO: 92. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to SEQ ID NO: 92. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 92.

[0237] In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to afragment of at least 600, at least 650, at least 675, or at least 700 amino acids of SEQ ID NO: 31. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of SEQ ID NO: 31. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of SEQ ID NO: 31. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO: 31. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to SEQ ID NO: 31. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 31.

[0238] An AAV6-derived capsid is (i) a capsid comprising at least a fragment of a wild-type AAV6 capsid, for example an AAV6-derived capsid may comprise at least 100, at least 200, at least 300, at least 400, at least 500, at least 600 or at least 700 amino acids of a wild-type AAV6 capsid, for example the AAV6 capsid of SEQ ID NO: 31 or (ii) a capsid having a sequence which has 100% identity to wild type AAV6 except for one or a small number of amino acid substitutions, and / or short insertions or replacements of up to about 15 amino acids in length. A capsid may further comprise one or more substitutions compared to a wild-type AAV6 capsid. A capsid may further comprise a fragment from the capsid of another AAV serotype. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

[0239] In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33. In some embodiments, the capsid comprises a polypeptide comprising an amino acid sequence of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

[0240] Transcription regulatory element

[0241] A polynucleotide of the invention may comprise a transcription regulatory element. A transcription regulatory element may comprise a promoter and / or an enhancer, and optionally other nucleotides. The transcription regulatory element may be operably linked to the transgene.

[0242] A transcription regulatory element directs the transcription of a transgene to which it is operably linked. For example, a promoter can regulate both the rate and efficiency of the transcription of an operably linked transgene. A promoter may also be operably linked toother regulatory sequences which enhance (“enhancer sequences") or repress (“repressor sequences'") promoter-dependent transcription of a transgene. An enhancer sequence may be an intron. The transcription regulatory elements may also include, without limitations, repressor and activator protein binding sites, and any other sequences of nucleotides known to one of skill in the art to act directly or indirectly to regulate the amount of transcription from the promoter, including e.g. attenuators, enhancer sequences, and silencers. The promoter is generally located near the transcription start site of the transgene to which it is operably linked, on the same strand and upstream of the DNA sequence (towards the 5’ region of the sense strand). As used herein, the term “operably linked' refers to a linkage of elements in a functional relationship. A transgene is “operably linked' when it is placed into a functional relationship with another nucleic acid sequence. For instance, a promoter or transcription regulatory sequence is operably linked to a transgene if it affects the transcription of the transgene. Optionally, the promoter or transcription regulatory sequence is 5’ of the transgene. Optionally, the promoter is immediately 5’ of the transgene, or it is separated from the transgene by another sequence such as an intron.

[0243] In some embodiments, the transcription regulatory element comprises a promoter. In some embodiments, the promoter drives expression of the transgene in ocular cells. In some embodiments the ocular cells are retinal and / or para-retinal cells, optionally including one or more cell types selected from retinal ganglion cells, amacrine cells, horizontal cells, bipolar cells, photoreceptor cells, cone cells, rod cells, Muller glial cells, retinal pigmented epithelium (RPE) cells and / or choroid cells. In some embodiments, the ocular cells are cells in the retinal pigment epithelial layer or cells in the choroidal capillary layer. In some embodiments the promoter drives expression in anterior ocular cell types such as cells of the ciliary body and / or iris, optionally in addition to the aforementioned retinal and para-retinal cells.

[0244] In some embodiments, the transcription regulatory element comprises at least one element selected from the group consisting of a promoter, an enhancer, a post-transcriptional regulatory element, an intron, a 3’ UTR, and a 5’ UTR.

[0245] In some embodiments, the polynucleotide comprises a CBA promoter. In some embodiments, the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is:

[0246] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, or at least 800 nucleotides of SEQ ID NO: 39;

[0247] (b) at least 95% identical to a fragment of at least 800 nucleotides of SEQ ID NO: 39; (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 39; or

[0248] (d) identical to SEQ ID NO: 39.

[0249] In some embodiments, the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 800 nucleotides of SEQ ID NO: 39. In some embodiments, the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 39. In some embodiments, the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 39. In some embodiments, the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 39.Woodchuck hepatitis post-transcriptional regulatory element (WPRE)

[0250] The polynucleotide of the invention may comprise a WPRE sequence. Optionally, the post-transcriptional regulatory element is able to increase expression of the transgene in HEK293T cells. Whether or not a post-transcriptional regulatory element is able to increase expression of the transgene may be determined by transfecting HEK293T cells with the polynucleotide, measuring the level of expression of the transgene, and comparing the level of expression with that of an equivalent (otherwise identical) polynucleotide lacking the post-transcriptional regulatory element.

[0251] In some embodiments, the WPRE comprises a polynucleotide comprising a nucleotide sequence which is:

[0252] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, or at least 550 nucleotides of SEQ ID NO: 40;

[0253] (b) at least 95% identical to a fragment of at least 550 nucleotides of SEQ ID NO: 40; (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 40; or

[0254] (d) identical to SEQ ID NO: 40.

[0255] In some embodiments, the WPRE comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 550 nucleotides of SEQ ID NO: 40. In some embodiments, the WPRE comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 40. In some embodiments, the WPRE comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 40. In some embodiments, the WPRE comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 40.

[0256] 5 ’ untranslated region (UTR)

[0257] The 5’ UTR is the section of a transgene which is immediately 5’ of the initiation codon. The 5’ UTR may comprise various elements which influence transcription of the transgene, for example upstream open reading frames (ORFs), internal ribosome entry sites (IRESs), microRNA binding sites, and secondary structure elements which regulate mRNA stability, pre-mRNA splicing, and translation initiation.

[0258] In some embodiments, the recombinant genome comprises a 5’ UTR.

[0259] Inverted terminal repeats (ITRs)

[0260] The term “ inverted terminal repeat' (ITR) refers to a nucleotide sequence located at the 5’ end (5’ ITR) and a nucleotide sequence located at the 3 ’end (3TTR) of a viral genome. An ITR comprises palindromic sequences and can fold over to form T-shaped hairpin structures that function as primers during initiation of DNA replication, in addition to performing other functions. The ITRs are required in cis for the vector genome replication and its packaging into the viral particles.In one embodiment of the invention, the recombinant genome comprises at least one ITR. In some embodiments, the recombinant genome comprises 1 or 2 ITRs. In some embodiments, the recombinant genome comprises a 5’ ITR and a 3’ ITR.

[0261] In some embodiments, the ITR sequences are from AAV1, AAV2, AAAV3, AAV4, AAV5, AAV6, AAV7, AAV 8, AAV9, or AAV10. In some embodiments of the invention, the ITR sequences are from AAV2 or AAV5. In some embodiments of the invention, the ITR sequences are from AAV2. In some embodiments, the 5’ ITR comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 93. In some embodiments, the 5’ ITR comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 93. In some embodiments, the 5’ ITR comprises a nucleotide sequence which is identical to SEQ ID NO: 93. In some embodiments, the 3’ ITR comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 94. In some embodiments, the 3’ ITR comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 94. In some embodiments, the 3’ ITR comprises a nucleotide sequence which is identical to SEQ ID NO: 94.

[0262] In some embodiments, the ITRs are truncated versions of a corresponding native ITR. In some embodiments, the 5’ ITR comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 88. In some embodiments, the 5’ ITR comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 88. In some embodiments, the 5’ ITR comprises a nucleotide sequence which is identical to SEQ ID NO: 88. In some embodiments, the 3’ ITR comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 89. In some embodiments, the 3’ ITR comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 89. In some embodiments, the 3’ ITR comprises a nucleotide sequence which is identical to SEQ ID NO: 89.

[0263] PolyA tail

[0264] As used herein, the term “ polyadenylation tail” (or polyA tail or polyA sequence) refers to a specific recognition sequence within the 3’ untranslated region (3’ UTR) of the gene, which is transcribed into a precursor mRNA molecule and guides the termination of gene transcription. The polyA sequence acts as a signal for the endonucleolytic cleavage of the newly formed precursor mRNA at its 3 ’-end, and for the addition of an RNA stretch consisting only of adenine bases to the 3’ -end. This is known as polyadenylation. The polyA tail is important for nuclear export, translation, and stability of mRNA.

[0265] In some embodiments, the recombinant genome further comprises a polyA sequence. In some embodiments, the recombinant genome comprises at least one, at least two, or at least three polyA sequences. In some embodiments, the polyA sequence(s) are selected from a bGH (bovine growth hormone) polyA, an hGH (human growth hormone) polyA, or an SV40 polyA. In some embodiments, the poly(A) sequences comprise, in 5’ to 3’ order, a bGH polyA, an hGH polyA, and an SV40 polyA.

[0266] In some embodiments, the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is:

[0267] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 100, at least 125, at least 150, at least 175, or at least 200 nucleotides of SEQ ID NO: 41;

[0268] (b) at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 41;(c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 41; or

[0269] (d) identical to SEQ ID NO: 41.

[0270] In some embodiments, the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 41. In some embodiments, the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 41. In some embodiments, the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 41. In some embodiments, the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 41.

[0271] In some embodiments, the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is:

[0272] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 100, at least 125, at least 150, at least 175, or at least 200 nucleotides of SEQ ID NO: 42; (b) at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 42; (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 42; or

[0273] (d) identical to SEQ ID NO: 42.

[0274] In some embodiments, the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 42. In some embodiments, the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 42. In some embodiments, the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 42. In some embodiments, the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 42.

[0275] In some embodiments, the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is:

[0276] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 100, at least 125, at least 150, or at least 175 nucleotides of SEQ ID NO: 43;

[0277] (b) at least 95% identical to a fragment of at least 175 nucleotides of SEQ ID NO: 43; (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 43; or

[0278] (d) identical to SEQ ID NO: 43.

[0279] In some embodiments, the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 175 nucleotides of SEQ ID NO: 43. In some embodiments, the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 43. In some embodiments, the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 43. In some embodiments, the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 43.

[0280] Complete vector genomeIn some embodiments, the present invention provides a vector comprising, in 5’ to 3’ order, a 5’ ITR, a promoter, a transgene encoding a C5 inhibitor, a WPRE sequence, a poly A sequence, and a 3 ’ ITR.

[0281] In some embodiments, the present invention provides a vector comprising, in 5’ to 3’ order, a 5’ ITR, a promoter, a transgene encoding a C5 inhibitor, a WPRE sequence, three poly A sequences, and a 3 ’ ITR.

[0282] Pharmaceutical composition

[0283] In some embodiments, there is provided a composition comprising the vector or the polypeptide of the invention and a pharmaceutically acceptable excipient. The term "pharmaceutically acceptable" means approved by a regulatory agency or recognized pharmacopeia such as European Pharmacopeia, for use in animals and / or humans. The term "excipient" refers to a diluent, adjuvant, carrier, or vehicle with which the therapeutic agent is administered.

[0284] Any suitable pharmaceutically acceptable carrier, diluent or excipient can be used in the preparation of a pharmaceutical composition (See e.g., Remington: The Science and Practice of Pharmacy, Alfonso R. Gennaro (Editor) Mack Publishing Company, April 1997).

[0285] Pharmaceutical compositions are typically sterile and stable under the conditions of manufacture and storage. Pharmaceutical compositions may be formulated as solutions (e.g. saline, dextrose solution, or buffered solution, or other pharmaceutically acceptable sterile fluids), microemulsions, liposomes, or other ordered structure suitable to accommodate a high product concentration (e.g. microparticles or nanoparticles). The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol or sorbitol, or salts such as sodium chloride in the composition.

[0286] Methods of medical treatment / medical uses

[0287] The terms “treatment'. “treating" and the like refer to obtaining a desired pharmacologic and / or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptoms thereof from appearing or worsening and / or may be therapeutic in terms of a partial or complete cure for a disease and / or symptom attributable to the disease.

[0288] In some embodiments, the method of treatment is a method of treating a complement-mediated disorder.Optionally, the complement-mediated disorder is an age-related inflammatory or autoinflammatory disease. Optionally, the complement-mediated disorder is a chronic inflammatory disease.

[0289] The term “ disorder associated with C5" refers to a disease, disorder, condition or syndrome which is caused, maintained or exacerbated, or whose signs and / or symptoms are caused, maintained or exacerbated, directly or indirectly, by complement system activity wherein the complement system activity can be reduced or stabilized or eliminated by inhibition of C5 activity. Such C5 activity can be inhibited by preventing, for example, cleavage of C5 precursor into C5a and C5b chains, formation of the membrane attack complex (MAC) and / or binding of the MAC to the surface of a target cell (e.g., a red blood cell).

[0290] In some embodiments, the method of treatment is a method of treating a disorder associated with C5 and / or a condition that would benefit from reducing complement-mediated inflammation.

[0291] In some embodiments, the complement-mediated disorder, the disorder associated with C5 and / or the condition that would benefit from reducing complement-mediated inflammation is an ocular disorder. In some embodiments, the ocular disorder is selected from age-related macular degeneration (AMD), geographic atrophy (GA), diabetic macular edema (DME), diabetic retinopathy, ocular angiogenesis (ocular neovascularization affecting choroidal, corneal or retinal tissue), uveitis, glaucoma, and neuromyelitis optica.

[0292] In some embodiments of the invention, treatment of an ocular disease may comprise an improvement in the quality of vision in a subject. For example, treatment of an ocular disease may comprise a reduction in blurry or fuzzy vision, reduction or elimination of a blind spot in the centre of vision, and / or reduction in the loss of central or peripheral vision. Treatment of an ocular disease may comprise an increase in fine detail of vision, an improvement in the ability to perceive faces, or an improvement in the ability to read. Treatment of an ocular disorder may be measured with a visual acuity test. A visual acuity test measures the ability of a subject to discern letters or symbols of various sizes from a distance. It is within the capabilities of the skilled person to perform a visual acuity test. In some embodiments, treatment of an ocular disorder comprises improving the performance of a subject in a visual acuity test.

[0293] In some embodiments, the present invention relates to a method of treating an ocular disease in a subject. In some embodiments, the method of treating an ocular disorder comprises a step of administration of a polynucleotide, viral particle, or composition of the invention to the subject. The method of treating an ocular disorder comprises a step of intravitreal administration of a polynucleotide, viral particle, or composition of the invention. In some embodiments, intravitreal administration comprises administration of the polynucleotide, viral particle or composition of the invention to the vitreous humour using a needle.

[0294] In some embodiments, the ocular disorder is a degenerative disorder or is a disorder mediated by inflammation, optionally resulting from an unbalanced complement system. In some embodiments, the ocular disease or disorder is one which is amenable to treatment through raising the local cell-extrinsic concentration of a therapeutic protein; in other words, a treatment which results in therapeutic gene expression in ocular cells but not necessarily inthe cells directly or potentially - at the time of administration of the polypeptide, vector or composition of the invention - affected by the disease or disorder. Symptoms which may be reduced or ameliorated include loss of vision, visual distortion, difficulty adapting to low light levels, crooked central vision, increase in haziness of central / overall vision, presence of drusen (tiny accumulations of extracellular material that build up on the retina), pigmentary changes, distorted vision in the form of metamorphopsia, in which a grid of straight lines appears wavy and parts of the grid may appear blank, exudative changes (hemorrhages in the eye, hard exudates, subretinal / sub-RPE / intraretinal fluid), slow recovery of visual function after exposure to bright light (photostress test), incipient and geographic atrophy, visual acuity drastically decreasing (two levels or more), e.g., 20 / 20 to 20 / 80, preferential hyperacuity perimetry changes (for wet AMD), blurred vision, gradual loss of central vision (for those with non-exudative macular degeneration), rapid onset of vision loss (often caused by leakage and bleeding of abnormal blood vessels in subjects with exudative macular degeneration), central scotomas (shadows or missing areas of vision), trouble discerning colours, specifically dark ones from dark ones and light ones from light ones, loss in contrast sensitivity, straight lines appearing curved in an Amsler grid.

[0295] In some embodiments, the ocular disorder is age-related macular degeneration (AMD). In a preferred embodiment, the ocular disorder is dry AMD. Dry AMD is characterised by an increase in apoptosis of the cells of the macula, in particular the cells of the retinal pigment epithelium layer, photoreceptor cells, and often cells in the choroidal capillary layer.

[0296] Progressive cell death of retinal pigment epithelial cells and photoreceptor cells in the macula may lead to geographic atrophy, which is accompanied by a worsening of central vision and a loss of the ability to see fine details. In some embodiments, administration of the polypeptide, vector, or composition of the invention leads to a reduction in the size of a geographic atrophy, or a slowing or stopping of the rate of expansion of geographic atrophy in a subject. In some embodiments, treating or preventing dry AMD comprises preventing or reducing the rate of progression of pathology or symptoms of dry AMD such as deterioration of central vision.

[0297] In some embodiments, the ocular disorder is diabetic retinopathy.

[0298] In some embodiments, the subject may carry one or two copies of a R885C / H polymorphism in C5 which adversely affects the subject’s response to treatment with Eculizumab.

[0299] Methods of the invention

[0300] The present invention provides a method of reducing drusen in the eye of a subject, the method comprising a step of intravitreal administration to the subject the vector or composition of the invention.

[0301] The present invention provides a method of preventing or slowing loss of vision in a subject, the method comprising a step of intravitreal administration to the subject the vector or composition of the invention.

[0302] In some embodiments, the subject may carry one or two copies of a R885C / H polymorphism in C5 which adversely affects the subject’s response to treatment with Eculizumab.Dosage

[0303] A suitable dosage of the vector, e.g. AAV particle, according to the present invention may be determined by a skilled practitioner.

[0304] A suitable total dose of the vector or composition to be administered according to the present invention as a whole is at least IxlO6vector genomes (vg). Preferably, a suitable total dose of AAV particles to be administered according to the present invention as a whole is in the range of about IxlO6to about IxlO14, in the range of about IxlO8to about IxlO14, in the range of about IxlO10to about IxlO14, or in the range of about IxlO10to about IxlO12vg of AAV particles. For example, where the AAV particles are administered to the eye, a suitable total dose of AAV particles to be administered according to the present invention as a whole is in the range of about IxlO6to about lxl014vg per eye, in the range of about IxlO8to about IxlO14vg per eye, in the range of about IxlO10to about IxlO14vg per eye, or in the range of about IxlO10to about IxlO12vg per eye of AAV particles.

[0305] In some embodiments, the total dose of AAV particles to be administered according to the present invention is 2.5 x IO10vg per eye, 7.5 x IO10vg per eye, or 2.5 x 1011vg per eye. In some embodiments, the total dose of AAV particles to be administered according to the present invention is between 2.5 x IO10vg per eye and 2.5 x 1011vg per eye. In some embodiments, the total dose of AAV particles to be administered according to the present invention is between 2.5 x IO10vg per eye and 7.5 x IO10vg per eye. In some embodiments, the total dose of AAV particles to be administered according to the present invention is between 7.5 x IO10vg per eye and 2.5 x 1011vg per eye.

[0306] As discussed in the summary of the invention, the vectors of the invention can be used to deliver (in the sense of in vivo expression) sufficient therapeutic protein to the retina, particularly the macula, for treating ocular diseases, even when low doses of the gene therapy are applied.

[0307] Alternatively, “normaD doses may be used, but when normal doses of the vector of the invention are applied, they may be more effective to treat disease.

[0308] In some embodiments, the vectors of the invention are part of a composition.

[0309] In some embodiments, the composition is a unit dose formulation of less than 4 x 1012vg, less than I x lO12vg, less than 9 x 1011vg, less than 8 x 1011vg, less than 7 x 1011vg, less than 6 x 1011vg, less than 5 x 1011vg, less than 4 x 1011vg, less than 3 x 1011vg, less than 2 x 1011vg, less than 1 x 1011vg, less than 8 x IO10vg, or less than 6 x IO10vg per eye. In some embodiments, the composition is a unit dose formulation of 3 x 1011vg or 2 x 1011vg per eye. In some embodiments, the composition is a unit dose formulation of about 1 x 109vg to about 4 x 1012vg per eye.

[0310] The term “unit dose formulation" refers to an amount of a composition that is to be administered as a dose. Thus, if the user intends for 5 x 1011vg to be administered, the unit dose formulation will be 5 x 1011vg. The unit dose formulation may initially be split between multiple containers, or may be comprised within a larger container, but a compositionconsisting of the unit dose formulation may be prepared for administration. For example, the presently claimed polynucleotides and viral particles may be used for intravitreal administration via an injection into the eye. In such cases, the syringe that is used for the injection may contain (z.e. the liquid within the syringe may consist of) the unit dose formulation.

[0311] In some embodiments, the step of intravitreal administration of the polynucleotide or composition is a step of intravitreal administration of the polynucleotide, viral particle or composition at a dose of less than 4 x 1012vg, less than 1 x 1012vg, less than 9 x 1011vg, less than 8 x 1011vg, less than 7 x 1011vg, less than 6 x 1011vg, less than 5 x 1011vg, less than 4 x 1011vg, less than 3 x 1011vg, less than 2 x 1011vg, less than 1 x 1011vg, less than 8 x IO10vg, or less than 6 x IO10vg per eye. In some embodiments, the step of intravitreal administration of the polynucleotide, viral particle or composition is a step of intravitreal administration of the polynucleotide, viral particle or composition at a dose of 3 x 1011vg or 2 x 1011vg per eye. In some embodiments, the step of intravitreal administration of the polynucleotide or composition is a step of intravitreal administration of the polynucleotide, viral particle or composition at a dose of between 1 x 109vg and 4 x 1012vg per eye.

[0312] Combination treatment with steroids

[0313] In some embodiments, the viral particle of the invention is administered in combination with a steroid. In some embodiments, the steroid is a corticosteroid. In some embodiments, the steroid is prednisolone. In some embodiments, the steroid is administered systemically. In some embodiments, the steroid is administered topically, for example the steroid may be administered to the eye. In some embodiments, the steroid is administered via periocular administration, intravitreal administration, subtenons administration, subconjunctival administration, or superchoroidal administration. In some embodiments, the steroid is administered via a drop. In some embodiments, the steroid is a glucocorticoid, for example an anti-inflammatory glucocorticoid. In some embodiments, the steroid is difluprednate, medrysone, loteprednol, prednisolone, fluocinolone, triamcinolone, rimexelone, dexamethasone, fluoromethoIone, or prednisone.

[0314] In some embodiments, the steroid is administered before, during, and / or after administration of the viral particle. In some embodiments, the steroid is administered before administration of the viral particle. In some embodiments, the steroid is administered during administration of the viral particle. In some embodiments, the steroid is administered after the viral particle. For example, the steroid maybe administered beginning a week after administration of the viral particle.

[0315] A A V production

[0316] The AAV particles according to the present invention may be produced by means of conventional methods and protocols. For example, viral particles of the invention may be produced according to any of the methods described in Clement and Grieger, Manufacturing of recombinant adeno-associated viral vectors for clinical trials, 2016, Molecular Therapy -Methods & Clinical Development, 3, 16002.Further guidance for the construction and production of viral vectors for use according to the disclosure can be found in Viral Vectors for Gene Therapy, Methods and Protocols. Series: Methods in Molecular Biology, Vol. 737. Merten and Al-Rubeai (Eds.); 2011 Humana Press (Springer); Gene Therapy. M. Giacca. 2010 Springer-Verlag; Heilbronn R. and Weger S. Viral Vectors for Gene Transfer: Current Status of Gene Therapeutics. In: Drug Delivery, Handbook of Experimental Pharmacology 197; M. Schafer-Korting (Ed.). 2010 Springer-Verlag; pp. 143-170; Adeno- Associated Virus: Methods and Protocols. R.O. Snyder and P. Mouillier (Eds). 2011 Humana Press (Springer); Biining H. et al. Recent developments in adeno-associated virus technology, 2008, J. Gene Med.; 10:717-733; Adenovirus: Methods and Protocols. M. Chillon and A. Bosch (Eds.); Third Edition. 2014 Humana Press (Springer).Informal Sequence Listing

[0317] SEQ Description SEQUENCE

[0318] ID NO

[0319] 1 CirpT 1 variant 1 EVRERGHTYVTKNVTVEDGACVYLRNVIPNGETKALNNPCVLSTCYAADRKVNSTLCPNI without signal GVDEGCHVEWTPDGVYPNCCPKHVCPSATASS

[0320] peptide

[0321] 2 CirpT 1 variant 2 DVQERGHTYVTKNVTVEDGACVYLRNVIPNGETKALNNPCVLSTCYAADRKVNSTLCPNI without signal GVDEGCHVEWTPDGVYPNCCPKHVCPSATASS

[0322] peptide

[0323] 3 CirpT2 without DVQERGHTYRTRNVTVEDGACVFERNVIPDGETKALNSPCVLSTCYAAAREVNSTLCPNI signal peptide GVEQGCRVEWTPVGEYPNCCPKHVCPTTS

[0324] 4 CirpT3 without DVQERGHTYVTKNVTVENGACVFERNVIPDGETKALNSPCVLSTCYAADRKVNSTLCPN signal peptide FGVAEGCHVEWTPDGEYPNCCPKHVCPAAPATS

[0325] 5 CirpT4 without DIQEHGHSYLTRNVTVENGACIFERNTLPDGETKALHDPCVIATCYAARREVNATLCRNF signal peptide GVDPGCRFHWRNDGVYPQCCPTQVCDGTD

[0326] 6 CirpT Consensus EXGHXYX{3}NVTXEXGACX{3}RNX{2}PXGETKALX{2}PCVX{2}TCYAXXRXVNXTLCXNX sequence without GVX{2}GCX{3}WX{3}GXYPXCCPX{2}VC

[0327] signal peptide

[0328] 7 CirpT Consensus EXGHXYX{3}NVTXEXGACX{3}RNX{2}PXGETKALX{2}PCVX{2}TCYAXXRXVNXTLCXNX sequence without GVX{2}GCX{3}WX{3}GXYPXCCPX{2}VC

[0329] signal peptide

[0330] 8 RaCH without EEVKTTPIPNHQCVNATCERKLDALGNAVITKCPQGCLCVVRGASNIVPANGTCFQLATT signal peptide KPPMAPGDNKDNKEEESN

[0331] 9 RaCI2 without EEANTTPISVKDQCANVTCRRTVDNRGKRHIDGCPPGCLCVLKGPDSKDNLDGTCYLLA signal peptide TTPKSTTTSTEQSFNMEE

[0332] 10 RaCI3 without SGESQSIQRKGQCEEVICHRKLNHLGERVTSGCPTGCLCVIREPDNVDNANGTCYALMS signal peptide SI I I I I I I PDGTTTSEEEE

[0333] 11 RaCI4 without QEPTTPLKAASQCSNVKCRRRFDHLGNSVTEGCPSGCLCVYQATGYNQEANGTCYELM signal peptide KTS I I I I I EGTPAQ

[0334] 12 RaCI consensus QCX{4}CXRX{5}GX{6}CPXGCLCVX{11}GTCX{2}L

[0335] without signal

[0336] peptide

[0337] 13 Human AHSG MKSLVLLLCLAQLWGCHS

[0338] signal peptide

[0339] 14 Native CirpT4 ATGCGTGCGTTCGTGGCTTTGTTCTGTACCCTCGTCGCTTTTGCGACGGTCATCTGT DNA sequence GACATCCAGGAGCACGGCCACTCGTACTTGACGAGGAATGTGACTGTCGAGAATGGT with signal GCCTGCATCTTTGAGCGTAACACGCTTCCTGATGGTGAAACCAAGGCACTTCATGAT peptide CCCTGTGTCATCGCCACCTGCTACGCTGCGAGGCGCGAAGTGAACGCCACCCTGTG CCGAAACTTCGGTGTCGACCCGGGCTGCAGGTTTCACTGGAGAAATGATGGTGTCTA TCCTCAGTGCTGCCCAACACAAGTCTGCGATGGCACGGAC

[0340] 15 CirpT4 codop 1 ATGAGAGCCTTTGTGGCCCTGTTCTGCACCCTGGTGGCCTTTGCCACAGTGATCTGT with signal GACATCCAGGAGCATGGCCACAGCTACCTGACCAGAAATGTGACAGTGGAGAATGG peptide AGCCTGCATCTTTGAGAGAAACACCCTGCCTGATGGAGAGACCAAGGCCCTGCATGA CCCCTGTGTGATAGCCACCTGCTATGCAGCCAGAAGAGAGGTCAATGCCACCCTGTG CAGAAACTTTGGAGTGGACCCAGGCTGCAGATTCCACTGGAGAAATGATGGAGTGTA CCCCCAGTGCTGCCCCACCCAAGTGTGTGATGGCACAGAC

[0341] 16 CirpT4 codop 2 ATGAGAGCTTTTGTGGCCCTCTTTTGCACCCTTGTGGCTTTTGCTACTGTGATTTGTG with signal ACATTCAGGAGCATGGCCACAGCTACCTCACAAGAAATGTGACAGTGGAAAATGGAG peptide CCTGCAI I I I I GAGAGAAACACTCTCCCTGATGGAGAAACAAAGGCCCTCCATGATCC CTGTGTGATTGCCACTTGCTATGCTGCCAGAAGGGAAGTGAATGCTACACTGTGCAG AAACTTTGGAGTGGACCCTGGCTGCAGATTCCATTGGAGAAATGATGGAGTGTACCC CCAATGCTGCCCCACACAAGTGTGTGATGGCACAGAT

[0342] 17 CirpT4 codop 3 ATGAGGGCTTTTGTGGCTCTGTTTTGTACCCTGGTGGCCTTTGCCACTGTGATCTGTG with signal ACATCCAGGAGCATGGCCACAGCTATCTGACCAGGAATGTGACTGTGGAGAATGGAG peptide CCTGCATCTTTGAGAGGAACACTCTGCCTGATGGAGAGACTAAGGCTCTGCATGACC

[0343]

[0344] CTTGTGTGATTGCCACCTGTTATGCTGCTAGAAGGGAGGTGAATGCTACCCTGTGTAGGAATTTTGGAGTGGACCCAGGCTGCAGGTTTCACTGGAGGAATGATGGAGTGTATC CACAGTGTTGTCCCACCCAAGTGTGTGATGGCACAGAC

[0345] CirpT4 codop 4 ATGAGAGCCTTTGTAGCCCTGTTTTGCACACTTGTAGCATTTGCCACAGTGATCTGTG with signal ACATTCAGGAACATGGGCACAGTTATCTGACTAGGAATGTCACAGTGGAGAATGGAG peptide CTTGTATCTTTGAGAGAAACACTCTGCCTGATGGAGAGACAAAGGCACTGCATGATC CATGTGTTATTGCAACCTGTTATGCTGCTAGAAGGGAGGTTAATGCTACCCTGTGTAG AAACTTTGGAGTAGACCCTGGCTGCAGATTTCATTGGAGGAATGATGGAGTTTACCCT CAGTGTTGTCCTACTCAAGTCTGTGATGGCACAGAC

[0346] CirpT4 codop 5 ATGCGGGCCTTCGTGGCTTTGTTCTGTACCCTCGTCGCTTTTGCGACCGTAATCTGT with signal GACATTCAGGAGCACGGCCACAGCTACCTGACGCGGAATGTGACTGTGGAGAATGG peptide TGCCTGCATATTTGAGAGAAACACACTGCCGGATGGGGAAACAAAGGCACTTCATGA TCCCTGTGTGATCGCCACCTGCTACGCTGCAAGGCGCGAAGTGAACGCCACCCTGT GTCGAAACTTCGGAGTCGACCCCGGATGCAGGTTTCATTGGAGAAATGACGGGGTTT ATCCTCAGTGTTGCCCAACACAAGTTTGCGATGGCACTGAC

[0347] CirpT4 codop 1 ATGAAGTCCCTGGTCCTGCTCCTTTGTCTTGCTCAACTCTGGGGCTGCCACTCAGAC plus AHSG signal ATCCAGGAGCATGGCCACAGCTACCTGACCAGAAATGTGACAGTGGAGAATGGAGC sequence CTGCATCTTTGAGAGAAACACCCTGCCTGATGGAGAGACCAAGGCCCTGCATGACCC CTGTGTGATAGCCACCTGCTATGCAGCCAGAAGAGAGGTCAATGCCACCCTGTGCAG AAACTTTGGAGTGGACCCAGGCTGCAGATTCCACTGGAGAAATGATGGAGTGTACCC CCAGTGCTGCCCCACCCAAGTGTGTGATGGCACAGAC

[0348] CirpT4 DNA GACATCCAGGAGCACGGCCACTCGTACTTGACGAGGAATGTGACTGTCGAGAATGGT sequence without GCCTGCATCTTTGAGCGTAACACGCTTCCTGATGGTGAAACCAAGGCACTTCATGAT signal peptide CCCTGTGTCATCGCCACCTGCTACGCTGCGAGGCGCGAAGTGAACGCCACCCTGTG CCGAAACTTCGGTGTCGACCCGGGCTGCAGGTTTCACTGGAGAAATGATGGTGTCTA TCCTCAGTGCTGCCCAACACAAGTCTGCGATGGCACGGAC

[0349] CirpT4 codop 1 GACATCCAGGAGCATGGCCACAGCTACCTGACCAGAAATGTGACAGTGGAGAATGG without signal AGCCTGCATCTTTGAGAGAAACACCCTGCCTGATGGAGAGACCAAGGCCCTGCATGA peptide CCCCTGTGTGATAGCCACCTGCTATGCAGCCAGAAGAGAGGTCAATGCCACCCTGTG CAGAAACTTTGGAGTGGACCCAGGCTGCAGATTCCACTGGAGAAATGATGGAGTGTA CCCCCAGTGCTGCCCCACCCAAGTGTGTGATGGCACAGAC

[0350] CirpT4 codop 2 GACATTCAGGAGCATGGCCACAGCTACCTCACAAGAAATGTGACAGTGGAAAATGGA without signal GCCTGCAI I I I I GAGAGAAACACTCTCCCTGATGGAGAAACAAAGGCCCTCCATGAT peptide CCCTGTGTGATTGCCACTTGCTATGCTGCCAGAAGGGAAGTGAATGCTACACTGTGC AGAAACTTTGGAGTGGACCCTGGCTGCAGATTCCATTGGAGAAATGATGGAGTGTAC CCCCAATGCTGCCCCACACAAGTGTGTGATGGCACAGAT

[0351] CirpT4 codop 3 GACATCCAGGAGCATGGCCACAGCTATCTGACCAGGAATGTGACTGTGGAGAATGGA without signal GCCTGCATCTTTGAGAGGAACACTCTGCCTGATGGAGAGACTAAGGCTCTGCATGAC peptide CCTTGTGTGATTGCCACCTGTTATGCTGCTAGAAGGGAGGTGAATGCTACCCTGTGT AGGAATTTTGGAGTGGACCCAGGCTGCAGGTTTCACTGGAGGAATGATGGAGTGTAT CCACAGTGTTGTCCCACCCAAGTGTGTGATGGCACAGAC

[0352] CirpT4 codop 4 GACATTCAGGAACATGGGCACAGTTATCTGACTAGGAATGTCACAGTGGAGAATGGA without signal GCTTGTATCTTTGAGAGAAACACTCTGCCTGATGGAGAGACAAAGGCACTGCATGAT peptide CCATGTGTTATTGCAACCTGTTATGCTGCTAGAAGGGAGGTTAATGCTACCCTGTGTA GAAACTTTGGAGTAGACCCTGGCTGCAGATTTCATTGGAGGAATGATGGAGTTTACC CTCAGTGTTGTCCTACTCAAGTCTGTGATGGCACAGAC

[0353] CirpT4 codop 5 GACATTCAGGAGCACGGCCACAGCTACCTGACGCGGAATGTGACTGTGGAGAATGG without signal TGCCTGCATATTTGAGAGAAACACACTGCCGGATGGGGAAACAAAGGCACTTCATGA peptide TCCCTGTGTGATCGCCACCTGCTACGCTGCAAGGCGCGAAGTGAACGCCACCCTGT GTCGAAACTTCGGAGTCGACCCCGGATGCAGGTTTCATTGGAGAAATGACGGGGTTT ATCCTCAGTGTTGCCCAACACAAGTTTGCGATGGCACTGAC AAV2-derived MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNG capsid LDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGR AVFQAKKRVLEPLGLVEEPVKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFG QTGDADSVPDPQPLGQPPAAPSGLGTNTMATGSGAPMADNNEGADGVGNSSGNWHC DSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEY

[0354]

[0355] QLPYVLGSAHQGCLPPFPADVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTNTPSGTTTQSRLQFSQAG ASDIRDQSRNWLPGPCYRQQRVSKTSADNNNSEYSWTGATKYHLNGRDSLVNPGPAM ASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQ RGNLALGETTRPARQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSP LMGGFGLKHPPPQILIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKR WNPEIQYTSNYNKSVNVDFTVDTNGVYSEPRPIGTRYLTRNL AAV2-derived MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNG capsid LDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGR AVFQAKKRVLEPFGLVEEGAKTAPTGKRIDDHFPKRKKARTEEDSKPSTSSDAEAGPSG SQQLQIPAQPASSLGADTMSAGGGGPLGDNNQGADGVGNASGDWHCDSTWMGDRVV TKSTRTWVLPSYNNHQYREIKSGSVDGSNANAYFGYSTPWGYFDFNRFHSHWSPRDW QRLINNYWGFRPRSLRVKIFNIQVKEVTVQDSTTTIANNLTSTVQVFTDDDYQLPYVVGNG TEGCLPAFPPQVFTLPQYGYATLNRDNTENPTERSSFFCLEYFPSKMLRTGNNFEFTYNF EEVPFHSSFAPSQNLFKLANPLVDQYLYRFVSTNNTGGVQFNKNLAGRYANTYKNWFPG PMGRTQGWNLGSGVNRASVSAFATTNRMELEGASYQVPPQPNGMTNNLQGSNTYALE NTMIFNSQPANPGTTATYLEGNMLITSESETQPVNRVAYNVGGQMLAHKFKSGDAPTTG TYNLQEIVPGSVWMERDVYLQGPIWAKIPETGAHFHPSPAMGGFGLKHPPPMMLIKNTP VPGNITSFSDVPVSSFITQYSTGQVTVEMEWELKKENSKRWNPEIQYTNNYNDPQFVDF APDSTGEYRTTRPIGTRYLTRPL AAV2-derived MAADGYLPDW LEDTLSEGIR QWWKLKPGPP PPKAAERHKD DSRGLVLPGY capsid KYLGPFNGLD KGEPVNEADA AALEHDKAYD

[0356] RQLDSGDNPY LKYNHADAEF QERLKEDTSF GGNLGRAVFQ AKKRVLEPLG LVEEPVKTAP GKKRPVEHSP VEPDSSSGTG KAGQQPARKR LNFGQTGDAD SVPDPQPLGQ PPAAPSGLGT NTMATGSGAP MADNNEGADG VGNSSGNWHC DSTWMGDRVI TTSTRTWALP TYNNHLYKQI SSQSGASNDN HYFGYSTPWG YFDFNRFHCH FSPRDWQRLI NNNWGFRPKR LNFKLFNIQV KEVTQNDGTT TIANNLTSTV QVFTDSEYQL PYVLGSAHQG CLPPFPADVF MVPQYGYLTL NNGSQAVGRS SFYCLEYFPS QMLRTGNNFT FSYTFEDVPF HSSYAHSQSL DRLMNPLIDQ YLYYLSRTNT PSGTTTQSRL QFSQAGASDI RDQSRNWLPG PCYRQQRVSK TSADNNNSEY SWTGATKYHL NGRDSLVNPG PAMASHKDDE EKFFPQSGVL IFGKQGSEKT NVDIEKVMIT DEEEIRTTNP VATEQYGSVS TNLQRGNLAI SDQTKHARQA ATADVNTQGV LPGMVWQDRD VYLQGPIWAK IPHTDGHFHP SPLMGGFGLK HPPPQILIKN TPVPANPSTT FSAAKFASFI TQYSTGQVSV EIEWELQKEN SKRWNPEIQY TSNYNKSVNV DFTVDTNGVY SEPRPIGTRY LTRNL AAV2-derived MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNG capsid LDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGR AVFQAKKRVLEPLGLVEEPVKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFG QTGDADSVPDPQPLGQPPAAPSGLGTNTMATGSGAPMADNNEGADGVGNSSGNWHC DSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEY QLPYVLGSAHQGCLPPFPADVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTG NNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYFLSRTNTPSGTTTQSRLQFSQAG ASDIRDQSRNWLPGPCYRQQRVSKTSADNNNSEFSWTGATKYHLNGRDSLVNPGPAM ASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQ RGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHP PPQILIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSN YNKSVNVDFTVDTNGVYSEPRPIGTRFLTRNL AAV6 capsid MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFN GLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLRYNHADAEFQERLQEDTSFGGNLG RAVFQAKKRVLEPFGLVEEGAKTAPGKKRPVEQSPQEPDSSSGIGKTGQQPAKKRLNFG QTGDSESVPDPQPLGEPPATPAAVGPTTMASGGGAPMADNNEGADGVGNASGNWHC DSTWLGDRVITTSTRTWALPTYNNHLYKQISSASTGASNDNHYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTTNDGVTTIANNLTSTVQVFSDSEY

[0357]

[0358] QLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLNRTQNQSGSAQNKDLLFSRGSP AGMSVQPKNWLPGPCYRQQRVSKTKTDNNNSNFTWTGASKYNLNGRESIINPGTAMAS HKDDKDKFFPMSGVMIFGKESAGASNTALDNVMITDEEEIKATNPVATERFGTVAVNLQS SSTDPATGDVHVMGALPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPP PQILIKNTPVPANPPAEFSATKFASFITQYSTGQVSVEIEWELQKENSKRWNPEVQYTSNY AKSANVDFTVDNNGLYTEPRPIGTRYLTRPL AAV6-derived MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFN capsid GLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLRYNHADAEFQERLQEDTSFGGNLG RAVFQAKKRVLEPLGLVEEGAKTAPGKKRPVEQSPQEPDSSSGIGKTGQQPAKKRLNFG QTGDSESVPDPQPLGEPPATPAAVGPTTMASGGGAPMADNNEGADGVGNASGNWHC DSTWLGDRVITTSTRTWALPTYNNHLYKQISSASTGASNDNHYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTTNDGVTTIANNLTSTVQVFSDSEY QLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGN NFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLNRTQNQSGSAQNKDLLFSRGSP AGMSVQPKNWLPGPCYRQQRVSKTKTDNNNSNFTWTGASKYNLNGRESIINPGTAMAS HKDDKDKFFPMSGVMIFGKESAGASNTALDNVMITDEEEIKATNPVATERFGTVAVNLQS SSTDPATGDVHVMGALPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPP PQILIKNTPVPANPPAEFSATKFASFITQYSTGQVSVEIEWELQKENSKRWNPEVQYTSNY AKSANVDFTVDNNGLYTEPRPIGTRYLTRPL AAV6-derived MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFN capsid GLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLRYNHADAEFQERLQEDTSFGGNLG RAVFQAKKRVLEPLGLVEEGAKTAPGKKRPVEQSPQEPDSSSGIGKTGQQPAKKRLNFG QTGDSESVPDPQPLGEPPATPAAVGPTTMASGGGAPMADNNEGADGVGNASGNWHC DSTWLGDRVITTSTRTWALPTYNNHLYKQISSASTGASNDNHYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTTNDGVTTIANNLTSTVQVFSDSEY QLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGN NFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLNRTQNQSGSAQNKDLLFSRGSP AGMSVQPKNWLPGPCYRQQRVSKTKTDNNNSNFTWTGASKYNLNGRESIINPGTAMAS HKDDKDKFFPMSGVMIFGKESAGASNTALDNVMITDEEEIKATNPVATERFGTVAVNLQN SSTDPATGDVHVMGALPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPP PQILIKNTPVPANPPAEFSATKFASFITQYSTGQVSVEIEWELQKENSKRWNPEVQYTSNY AKSANVDFTVDNNGLYTEPRPIGTRYLTRPL AAV6-derived MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFN capsid GLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLRYNHADAEFQERLQEDTSFGGNLG RAVFQAKKRVLEPFGLVEEGAKTAPGKKRPVEQSPQEPDSSSGIGKTGQQPAKKRLNFG QTGDSESVPDPQPLGEPPATPAAVGPTTMASGGGAPMADNNEGADGVGNASGNWHC DSTWLGDRVITTSTRTWALPTYNNHLYKQISSASTGASNDNHYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNVQVKEVTTNDGVTTIANNLTSTVQVFSDSE YQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTG NNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLNRTQDQSGSAQNKDLLFSRG SPAGMSVQPKNWLPGPCYRQQRVSKTKTDNNNSNFTWTGASKYNLNGRESIINPGTAM ASHKDDKNKFFPMSGVMIFGKESAGASNTALDNVMITDEEEIKATNPVATERFGTVAVNL QSSSTDPATGDVHVMGALPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKN PPPQILIKNTPVPANPPAEFSATKFASFITQYSTGQVSVEIEWELQKENSKRWNPEVQYTS NYAKSANVDFTVDNNGLYTEPRPIGTRYLTRPL AAV2 capsid MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNG LDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGR AVFQAKKRVLEPLGLVEEPVKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFG QTGDADSVPDPQPLGQPPAAPSGLGTNTMATGSGAPMADNNEGADGVGNSSGNWHC DSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEY QLPYVLGSAHQGCLPPFPADVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTG NNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTNTPSGTTTQSRLQFSQAG ASDIRDQSRNWLPGPCYRQQRVSKTSADNNNSEYSWTGATKYHLNGRDSLVNPGPAM ASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQ RGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHP PPQILIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSN

[0359]

[0360] YNKSVNVDFTVDTNGVYSEPRPIGTRYLTRNLRaCI5 without TAEATLSINGGDMCIEKTCNRSIDAAGKKVIAGCPGGCLCVFNVSDVTYPANGTCYQLAT signal peptide TTTNRPGAVMERER

[0361] RaCI7 without SGESQSIQRKGQCEEVTCHRTLNHLGVAVTSGCPSGCLCVISAPDSAVNVNGTCYQLMG signal peptide STSTTTSSTPSSEDQE

[0362] OmCI without DSESDCTGSEPVDAFQAFSEGKEAYVLVRSTDPKARDCLKGEPAGEKQDNTLPVMMTF signal peptide KNGTDWASTDWTFTLDGAKVTATLGNLTQNREVVYDSQSHHCHVDKVEKEVPDYEMW MLDAGGLEVEVECCRQKLEELASGRNQMYPHLKDC CBA promoter CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCC CATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTG ACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTAT CATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCAT TATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACTCGAGGC CACGTTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTA TTTATTTTTTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGGGCGCGC GCCAGGCGGGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTG CGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGCGG CGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGGGAGCGGGATCA GCCACCGCGGTGGCGGCCTAGAGTCGACGAGGAACTGAAAAACCAGAAAGTTAACT GGTAAGTTTAGTC 1 1 1 1 1 GTCTTTTATTTCAGGTCCCGGATCCGGTGGTGGTGCAAAT CAAAGAACTGCTCCTCAGTGGATGTTGCCTTTACTTCTAGGCCTGTACGGAAGTGTTA CTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCACCGGTCCGGAATTC CCGGGATATCGTCGACCCACGCGTCCGGGCCCCACGCTGCGCACCCGCGGGTTTG CT WPRE AATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGC TCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCC GTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAG TTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACC CCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTC CCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGAC AGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGT CCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCT GCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCG GCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTT TGGGCCGCCTCCCCGC

[0363] bGH polyA CTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGAC CCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCAT TGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGG GGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGA

[0364] hGH polyA GGGTGGCATCCCTGTGACCCCTCCCCAGTGCCTCTCCTGGCCCTGGAAGTTGCCAC TCCAGTGCCCACCAGCCTTGTCCTAATAAAATTAAGTTGCATCATTTTGTCTGACTAG GTGTCCTTCTATAATATTATGGGGTGGAGGGGGGTGGTATGGAGCAAGGGGCAAGTT GGGAAGACAACCTGTAGGGCCTGCGGGGTCTATTGGGAACCAAGCTGGAGTGCAGT GGCACAATCTT SV40 polyA TTGGGCACTGACAATTCCGTGGTGTTTATTTGTGAAATTTGTGATGCTATTGCTTTATT TGTAACCATCTAGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTA TAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAG GGGGAGATGTGGGAGG 1 1 1 1 1 1

[0365] Native CirpTI GAAGTCCGGGAGCGCGGCCATACCTACGTGACCAAAAATGTGACGGTGGAAGATGG DNA sequence TGCCTGTGTGTATCTACGCAACGTCATTCCGAATGGTGAAACCAAAGCACTGAACAAC without signal CCATGCGTCCTTTCCACATGCTACGCGGCTGACCGTAAAGTGAACTCGACTCTCTGC peptide CCGAACATTGGAGTTGATGAGGGCTGCCATGTGGAGTGGACCCCCGATGGTGTATA CCCGAACTGCTGCCCGAAGCACGTGTGCCCTTCGGCCACCGCCAGTTCT

[0366] Native CirpT2 GACGTCCAAGAGCGCGGTCACACCTACAGGACCAGAAATGTGACTGTCGAAGATGG DNA sequence AGCCTGCGTCTTTGAGCGCAACGTGATACCGGATGGTGAAACTAAGGCGCTCAACAG

[0367]

[0368] CCCCTGCGTCCTTTCCACCTGCTACGCTGCGGCCCGGGAAGTGAATTCGACGCTCTwithout signal GCCCGAACATTGGAGTCGAGCAGGGCTGCAGGGTGGAGTGGACCCCCGTCGGAGA peptide GTATCCCAACTGCTGCCCGAAGCATGTGTGCCCCACCACCTCT

[0369] Native CirpT3 GACGTCCAGGAGCGCGGCCACACCTACGTGACCAAAAATGTGACGGTCGAAAACGG DNA sequence TGCCTGCGTGTTTGAACGCAACGTCATTCCGGATGGTGAAACCAAAGCACTGAACAG without signal CCCATGCGTCCTTTCCACATGCTACGCAGCTGACCGTAAAGTGAACTCGACTCTCTG peptide CCCGAACTTCGGAGTTGCGGAGGGCTGCCATGTGGAGTGGACCCCCGATGGTGAAT ACCCGAACTGCTGCCCGAAGCATGTGTGCCCTGCGGCCCCTGCTACTTCT

[0370] Native RaCI1 GAAGAGGTTAAGACTACACCTATACCAAATCACCAGTGTGTTAACGCGACTTGTGAGC DNA sequence GGAAACTCGACGCATTGGGAAACGCAGTGATCACCAAATGTCCACAGGGTTGCTTAT without signal GTGTCGTCAGAGGAGCCAGCAATATCGTTCCTGCAAATGGGACATGCTTCCAGCTAG peptide CGACAACCAAACCTCCCATGGCGCCTGGGGATAACAAAGACAATAAGGAGGAGGAAT CAAAC

[0371] Native RaCI2 GAAGAAGCGAACACTACGCCCATTTCAGTGAAAGACCAGTGCGCAAATGTGACATGC DNA sequence CGCCGTACTGTCGACAATAGAGGGAAAAGACATATTGATGGCTGCCCACCGGGTTGC without signal CTATGTGTTCTCAAGGGACCTGATAGTAAAGATAACCTGGATGGCACATGCTACCTTC peptide TGGCCACGACCCCAAAGTCCACTACCACTTCTACTGAACAGTCGTTCAACATGGAAG AG

[0372] Native RaCI3 AGTGGTGAGAGTCAATCCATCCAGAGGAAAGGCCAGTGTGAGGAAGTGATATGCCAC DNA sequence CGTAAACTCAACCACCTCGGTGAACGAGTGACAAGCGGTTGCCCCACTGGCTGTTTA without signal TGTGTCATCAGAGAACCCGATAATGTGGACAACGCAAATGGGACGTGCTATGCACTA peptide ATG

[0373] Native RaCI4 CAAGAGCCAACTACACCCCTGAAAGCGGCGAGCCAATGTTCCAATGTGAAATGTCGT DNA sequence CGTCGTTTCGACCACTTGGGCAACTCAGTGACTGAAGGGTGTCCGTCCGGCTGTTTA without signal TGTGTATACCAGGCAACTGGTTATAACCAAGAGGCAAATGGAACGTGCTACGAACTG peptide ATGAAAACTTCGACTACCACTACGACCGAGGGCACACCAGCACAG

[0374] Native RaCI5 ACTGCGGAGGCGACTTTGTCCATCAACGGAGGAGATATGTGCATCGAAAAGACATGC DNA sequence AACCGTTCCATCGACGCGGCAGGCAAGAAAGTGATAGCCGGTTGCCCAGGAGGTTG without signal TTTGTGTGTCTTCAACGTATCCGACGTCACTTATCCAGCAAACGGAACGTGCTACCAA peptide CTTGCAACCACCACCACCAATCGCCCCGGTGCTGTGATGGAAAGAGAGCGA Native RaCI7 AGTGGTGAGAGTCAATCCATCCAGAGGAAAGGCCAGTGTGAGGAAGTGACATGCCA DNA sequence CCGTACACTCAACCACCTCGGTGTAGCAGTGACAAGCGGTTGCCCGTCTGGCTGTTT without signal ATGTGTCATCAGCGCACCCGATAGTGCAGTCAACGTAAACGGGACGTGCTATCAACT peptide AATGGGTTCCACCAGCACCACGACAAGCAGTACACCCTCGTCGGAGGACCAAGAA Native OmCI DNA GACAGCGAAAGCGACTGCACTGGAAGCGAACCTGTTGACGCCTTCCAAGCTTTCAGT sequence without GAGGGCAAAGAGGCATATGTCCTGGTGAGGTCCACGGATCCCAAAGCGAGGGACTG signal peptide CTTGAAAGGAGAACCAGCCGGAGAAAAGCAGGACAACACGTTGCCGGTGATGATGA CGTTTAAGAATGGCACAGACTGGGCTTCAACCGATTGGACGTTTACTTTGGACGGCG CAAAGGTAACGGCAACCCTTGGTAACCTAACCCAAAATAGGGAAGTGGTCTACGACT CGCAAAGTCATCACTGCCACGTTGACAAGGTCGAGAAGGAAGTTCCAGATTATGAGA TGTGGATGCTCGATGCGGGAGGGCTTGAAGTGGAAGTCGAGTGCTGCCGTCAAAAG CTTGAAGAGTTGGCGTCTGGCAGGAACCAAATGTATCCCCATCTCAAGGACTGC

[0375] Codop CirpTI GAAGTCAGGGAGAGAGGCCACACCTATGTGACCAAGAATGTGACAGTGGAGGATGG DNA sequence AGCCTGTGTGTACCTGAGAAATGTGATCCCCAATGGAGAGACCAAGGCCCTGAACAA without signal CCCCTGTGTGCTGAGCACCTGCTATGCAGCTGACAGAAAAGTCAACAGCACCCTGTG peptide CCCCAACATTGGAGTGGATGAGGGCTGCCATGTGGAGTGGACCCCTGATGGAGTGT ACCCCAACTGCTGCCCCAAGCATGTGTGCCCCAGTGCCACAGCCAGCAGC

[0376] Codop CirpT2 GATGTGCAGGAGAGAGGCCACACCTACAGAACCAGAAATGTGACAGTGGAGGATGG DNA sequence AGCCTGTGTGTTTGAGAGAAATGTGATCCCTGATGGAGAGACCAAGGCCCTGAACAG without signal CCCCTGTGTGCTGAGCACCTGCTATGCAGCTGCCAGAGAAGTGAACAGCACCCTGT peptide GCCCCAACATTGGAGTGGAGCAGGGCTGCAGAGTGGAGTGGACCCCTGTGGGAGA GTACCCCAACTGCTGCCCCAAGCATGTGTGCCCCACCACCAGC

[0377] Codop CirpT3 GATGTGCAGGAGAGAGGACACACCTATGTGACCAAGAATGTGACAGTGGAGAATGG DNA sequence AGCCTGTGTGTTTGAGAGAAATGTGATCCCTGATGGAGAGACCAAGGCCCTGAACAG without signal CCCCTGTGTGCTGAGCACCTGCTATGCAGCTGACAGAAAGGTCAACAGCACCCTGTG peptide CCCCAACTTTGGTGTGGCAGAGGGCTGCCATGTGGAGTGGACCCCTGATGGAGAGT

[0378]

[0379] ACCCCAACTGCTGCCCCAAGCATGTGTGCCCTGCTGCCCCTGCCACCAGCCodop RaCI1 GAGGAGGTGAAGACCACCCCCATCCCCAACCACCAGTGTGTGAATGCCACCTGTGA DNA sequence GAGAAAGCTGGATGCCCTGGGCAATGCAGTGATCACCAAGTGCCCCCAGGGCTGCC without signal TGTGTGTGGTGAGAGGAGCCAGCAACATTGTGCCAGCCAATGGCACCTGCTTCCAAC peptide TGGCCACCACCAAGCCCCCAATGGCCCCTGGAGACAACAAGGACAACAAGGAGGAG GAGAGCAAC

[0380] Codop RaCI2 GAGGAGGCCAACACCACCCCCATCAGTGTGAAGGACCAGTGTGCCAATGTGACCTG DNA sequence CAGAAGGACAGTGGACAACAGAGGCAAGAGACACATAGATGGCTGCCCCCCAGGCT without signal GCCTGTGTGTGCTGAAGGGCCCAGACAGCAAGGACAACCTGGATGGCACCTGCTAC peptide CTGCTGGCCACCACCCCCAAGAGCACCACAACCAGCACAGAGCAGAGCTTCAACAT GGAGGAG

[0381] Codop RaCI3 AGTGGAGAGAGCCAGAGCATCCAGAGAAAGGGCCAGTGTGAGGAGGTGATCTGCCA DNA sequence CAGAAAGCTGAACCACCTGGGAGAGAGGGTGACATCAGGTTGCCCAACAGGCTGTC without signal TCTGTGTAATCAGAGAGCCAGACAATGTGGACAATGCCAATGGCACCTGCTATGCCC peptide TGATGAGCAGCACCACAACCACAACCACCACTCCTGATGGCACCACCACCAGTGAGG AGGAGGAG

[0382] Codop RaCI4 CAGGAGCCCACCACCCCCCTGAAGGCTGCCAGCCAGTGCAGCAATGTGAAGTGCAG DNA sequence GAGAAGGTTTGACCACCTGGGCAACAGTGTGACAGAGGGCTGCCCCAGTGGCTGCC without signal TGTGTGTGTACCAAGCCACAGGCTACAACCAGGAGGCCAATGGCACCTGCTATGAGC peptide TGATGAAGACCAGCACCACAACCACTACAGAGGGCACCCCAGCCCAG

[0383] Codop RaCI5 ACAGCTGAGGCCACCCTGAGCATCAATGGAGGAGACATGTGCATTGAGAAGACCTG DNA sequence CAACAGAAGCATTGATGCTGCTGGCAAGAAGGTCATTGCTGGCTGCCCTGGAGGCT without signal GCCTGTGTGTGTTCAATGTCAGTGATGTGACCTACCCTGCCAATGGCACCTGCTACC peptide AGCTGGCCACCACAACCACTAACAGACCTGGAGCTGTGATGGAGAGAGAGAGG Codop RaCI7 AGTGGAGAGAGCCAGAGCATCCAGAGGAAGGGCCAGTGTGAGGAGGTGACCTGCC DNA sequence ACAGAACCCTGAACCACCTGGGAGTGGCAGTGACCAGTGGCTGCCCCAGTGGCTGC without signal CTGTGTGTGATCAGTGCCCCAGACAGTGCAGTGAATGTGAATGGCACCTGCTACCAG peptide CTGATGGGCAGCACCAGCACAACCACTAGCAGCACCCCCAGCAGTGAGGACCAGGA G

[0384] Codop OmCI GACAGTGAGAGTGACTGCACAGGCAGTGAGCCTGTGGATGCCTTCCAAGCCTTCAGT DNA sequence GAGGGCAAGGAGGCCTATGTGCTGGTCAGAAGCACAGACCCCAAGGCCAGAGACTG without signal CCTGAAGGGAGAGCCAGCAGGAGAGAAGCAGGACAACACCCTGCCAGTGATGATGA peptide CCTTCAAGAATGGCACAGACTGGGCCAGCACAGACTGGACCTTCACCCTGGATGGA GCCAAGGTCACAGCCACCCTGGGCAACCTGACCCAGAACAGAGAGGTGGTGTATGA CAGCCAGAGCCACCACTGCCATGTGGACAAGGTGGAGAAGGAGGTGCCTGACTATG AGATGTGGATGCTGGATGCTGGAGGCCTGGAGGTGGAGGTGGAGTGCTGCAGACAG AAGCTGGAGGAGCTGGCCAGTGGCAGAAACCAGATGTACCCCCACCTGAAGGACTG C

[0385] Native CirpTI ATGGCGACACTAATCGCCGCGAGAACGAAGAGAAAAGCCCCGAGAGTGCGCATCTT DNA sequence CGAAGTCCGGGAGCGCGGCCATACCTACGTGACCAAAAATGTGACGGTGGAAGATG with signal GTGCCTGTGTGTATCTACGCAACGTCATTCCGAATGGTGAAACCAAAGCACTGAACA peptide ACCCATGCGTCCTTTCCACATGCTACGCGGCTGACCGTAAAGTGAACTCGACTCTCT GCCCGAACATTGGAGTTGATGAGGGCTGCCATGTGGAGTGGACCCCCGATGGTGTA TACCCGAACTGCTGCCCGAAGCACGTGTGCCCTTCGGCCACCGCCAGTTCT

[0386] Native CirpT2 ATGCGCACTCTTGTGGCTTCGCTATGTGTTTTCGCGGTGTTTAGTGCCGTCTGCTGC DNA sequence GACGTCCAAGAGCGCGGTCACACCTACAGGACCAGAAATGTGACTGTCGAAGATGG with signal AGCCTGCGTCTTTGAGCGCAACGTGATACCGGATGGTGAAACTAAGGCGCTCAACAG peptide CCCCTGCGTCCTTTCCACCTGCTACGCTGCGGCCCGGGAAGTGAATTCGACGCTCT GCCCGAACATTGGAGTCGAGCAGGGCTGCAGGGTGGAGTGGACCCCCGTCGGAGA GTATCCCAACTGCTGCCCGAAGCATGTGTGCCCCACCACCTCT

[0387] Native CirpT3 ATGCGCACTCTCGGGGTTTCTCTTTTCGTTCTTGTCGGGATTAGTGCCGTTTACTGTG DNA sequence ACGTCCAGGAGCGCGGCCACACCTACGTGACCAAAAATGTGACGGTCGAAAACGGT with signal GCCTGCGTGTTTGAACGCAACGTCATTCCGGATGGTGAAACCAAAGCACTGAACAGC peptide CCATGCGTCCTTTCCACATGCTACGCAGCTGACCGTAAAGTGAACTCGACTCTCTGC CCGAACTTCGGAGTTGCGGAGGGCTGCCATGTGGAGTGGACCCCCGATGGTGAATA CCCGAACTGCTGCCCGAAGCATGTGTGCCCTGCGGCCCCTGCTACTTCT

[0388]

[0389] Codop CirpTI ATGGCCACCCTGATTGCAGCAAGAACCAAGAGAAAGGCCCCCAGAGTGAGAATCTTC DNA sequence GAAGTCAGGGAGAGAGGCCACACCTATGTGACCAAGAATGTGACAGTGGAGGATGG with signal AGCCTGTGTGTACCTGAGAAATGTGATCCCCAATGGAGAGACCAAGGCCCTGAACAA peptide CCCCTGTGTGCTGAGCACCTGCTATGCAGCTGACAGAAAAGTCAACAGCACCCTGTG CCCCAACATTGGAGTGGATGAGGGCTGCCATGTGGAGTGGACCCCTGATGGAGTGT ACCCCAACTGCTGCCCCAAGCATGTGTGCCCCAGTGCCACAGCCAGCAGC

[0390] Codop CirpT2 ATGAGAACCCTGGTGGCCAGCCTGTGTGTGTTTGCTGTGTTCAGTGCAGTGTGCTGT DNA sequence GATGTGCAGGAGAGAGGCCACACCTACAGAACCAGAAATGTGACAGTGGAGGATGG with signal AGCCTGTGTGTTTGAGAGAAATGTGATCCCTGATGGAGAGACCAAGGCCCTGAACAG peptide CCCCTGTGTGCTGAGCACCTGCTATGCAGCTGCCAGAGAAGTGAACAGCACCCTGT GCCCCAACATTGGAGTGGAGCAGGGCTGCAGAGTGGAGTGGACCCCTGTGGGAGA GTACCCCAACTGCTGCCCCAAGCATGTGTGCCCCACCACCAGC

[0391] Codop CirpT3 ATGAGAACCCTGGGAGTGAGCCTGTTTGTGCTGGTGGGCATCAGTGCAGTGTACTGT DNA sequence GATGTGCAGGAGAGAGGACACACCTATGTGACCAAGAATGTGACAGTGGAGAATGG with signal AGCCTGTGTGTTTGAGAGAAATGTGATCCCTGATGGAGAGACCAAGGCCCTGAACAG peptide CCCCTGTGTGCTGAGCACCTGCTATGCAGCTGACAGAAAGGTCAACAGCACCCTGTG CCCCAACTTTGGTGTGGCAGAGGGCTGCCATGTGGAGTGGACCCCTGATGGAGAGT ACCCCAACTGCTGCCCCAAGCATGTGTGCCCTGCTGCCCCTGCCACCAGC

[0392] Native RaCI1 ATGAACGCTATGCTAGTCC 1 1 1 1 IAI CGCCTCTGCCTTGTTTATAAGTGAGCACAACA DNA sequence CGGAAGAGGTTAAGACTACACCTATACCAAATCACCAGTGTGTTAACGCGACTTGTGA with signal GCGGAAACTCGACGCATTGGGAAACGCAGTGATCACCAAATGTCCACAGGGTTGCTT peptide ATGTGTCGTCAGAGGAGCCAGCAATATCGTTCCTGCAAATGGGACATGCTTCCAGCT AGCGACAACCAAACCTCCCATGGCGCCTGGGGATAACAAAGACAATAAGGAGGAGG AATCAAAC

[0393] Native RaCI2 ATGAACGCTGTGACAGTCCTCGCTTTTACCGCATTTGCCCTTATTGTACATGACTGCT DNA sequence ACAGCGAAGAAGCGAACACTACGCCCATTTCAGTGAAAGACCAGTGCGCAAATGTGA with signal CATGCCGCCGTACTGTCGACAATAGAGGGAAAAGACATATTGATGGCTGCCCACCGG peptide GTTGCCTATGTGTTCTCAAGGGACCTGATAGTAAAGATAACCTGGATGGCACATGCTA CCTTCTGGCCACGACCCCAAAGTCCACTACCACTTCTACTGAACAGTCGTTCAACATG GAAGAG

[0394] Native RaCI3 ATGGCTGCACTGAACGGTCTCGTACTTTTGCTGCTTACCATTTCTGCCATGTTCATAA DNA sequence GTGAGTGTTACAGTAGTGGTGAGAGTCAATCCATCCAGAGGAAAGGCCAGTGTGAGG with signal AAGTGATATGCCACCGTAAACTCAACCACCTCGGTGAACGAGTGACAAGCGGTTGCC peptide CCACTGGCTGTTTATGTGTCATCAGAGAACCCGATAATGTGGACAACGCAAATGGGA CGTGCTATGCACTAATG

[0395] Native RaCI4 ATGTCTGCATTCAACATTTTCGCACTAGTGGTTGTCGTCTGTGCCTTGATGATAAATG DNA sequence AGTGCTGCACTAGCCAAGAGCCAACTACACCCCTGAAAGCGGCGAGCCAATGTTCCA with signal ATGTGAAATGTCGTCGTCGTTTCGACCACTTGGGCAACTCAGTGACTGAAGGGTGTC peptide CGTCCGGCTGTTTATGTGTATACCAGGCAACTGGTTATAACCAAGAGGCAAATGGAA CGTGCTACGAACTGATGAAAACTTCGACTACCACTACGACCGAGGGCACACCAGCAC AG

[0396] Native RaCI5 ATGAACGCTGTGATAGTTCTGTGCGTTACCATCTCTGCCGTACTGATACATCAGTGCT DNA sequence ACAGCACTGCGGAGGCGACTTTGTCCATCAACGGAGGAGATATGTGCATCGAAAAGA with signal CATGCAACCGTTCCATCGACGCGGCAGGCAAGAAAGTGATAGCCGGTTGCCCAGGA peptide GGTTGTTTGTGTGTCTTCAACGTATCCGACGTCACTTATCCAGCAAACGGAACGTGCT ACCAACTTGCAACCACCACCACCAATCGCCCCGGTGCTGTGATGGAAAGAGAGCGA

[0397] Native RaCI7 ATGGCTGCACTGAACGGTCTCGTACTTTTGCTGCTTACCATTTCTGCTATGTTCATAA DNA sequence GTGAGTGTTACAGTAGTGGTGAGAGTCAATCCATCCAGAGGAAAGGCCAGTGTGAGG with signal AAGTGACATGCCACCGTACACTCAACCACCTCGGTGTAGCAGTGACAAGCGGTTGCC peptide CGTCTGGCTGTTTATGTGTCATCAGCGCACCCGATAGTGCAGTCAACGTAAACGGGA CGTGCTATCAACTAATGGGTTCCACCAGCACCACGACAAGCAGTACACCCTCGTCGG AGGACCAAGAA

[0398] Native OmCI DNA ATGCTGGTTTTGGTGACCCTGATTTTCTCCTTTTCTGCGAACATCGCATATGCTGACA sequence with GCGAAAGCGACTGCACTGGAAGCGAACCTGTTGACGCCTTCCAAGCTTTCAGTGAGG signal peptide GCAAAGAGGCATATGTCCTGGTGAGGTCCACGGATCCCAAAGCGAGGGACTGCTTG

[0399] AAAGGAGAACCAGCCGGAGAAAAGCAGGACAACACGTTGCCGGTGATGATGACGTT

[0400]

[0401] TAAGAATGGCACAGACTGGGCTTCAACCGATTGGACGTTTACTTTGGACGGCGCAAAGGTAACGGCAACCCTTGGTAACCTAACCCAAAATAGGGAAGTGGTCTACGACTCGCA AAGTCATCACTGCCACGTTGACAAGGTCGAGAAGGAAGTTCCAGATTATGAGATGTG GATGCTCGATGCGGGAGGGCTTGAAGTGGAAGTCGAGTGCTGCCGTCAAAAGCTTG AAGAGTTGGCGTCTGGCAGGAACCAAATGTATCCCCATCTCAAGGACTGC

[0402] Codop RaCI1 ATGAATGCCATGCTGGTGCTGTTCATTGCCAGTGCCCTGTTCATCAGTGAGCACAAC DNA sequence ACAGAGGAGGTGAAGACCACCCCCATCCCCAACCACCAGTGTGTGAATGCCACCTGT with signal GAGAGAAAGCTGGATGCCCTGGGCAATGCAGTGATCACCAAGTGCCCCCAGGGCTG peptide CCTGTGTGTGGTGAGAGGAGCCAGCAACATTGTGCCAGCCAATGGCACCTGCTTCCA ACTGGCCACCACCAAGCCCCCAATGGCCCCTGGAGACAACAAGGACAACAAGGAGG AGGAGAGCAAC

[0403] Codop RaCI2 ATGAATGCTGTGACAGTGCTGGCCTTCACAGCCTTTGCCCTGATAGTGCATGACTGC DNA sequence TACAGTGAGGAGGCCAACACCACCCCCATCAGTGTGAAGGACCAGTGTGCCAATGT with signal GACCTGCAGAAGGACAGTGGACAACAGAGGCAAGAGACACATAGATGGCTGCCCCC peptide CAGGCTGCCTGTGTGTGCTGAAGGGCCCAGACAGCAAGGACAACCTGGATGGCACC TGCTACCTGCTGGCCACCACCCCCAAGAGCACCACAACCAGCACAGAGCAGAGCTT CAACATGGAGGAG

[0404] Codop RaCI3 ATGGCAGCCCTGAATGGCCTGGTGCTGCTGCTGCTGACCATCAGTGCCATGTTCATC DNA sequence TCTGAGTGCTACAGCAGTGGAGAGAGCCAGAGCATCCAGAGAAAGGGCCAGTGTGA with signal GGAGGTGATCTGCCACAGAAAGCTGAACCACCTGGGAGAGAGGGTGACATCAGGTT peptide GCCCAACAGGCTGTCTCTGTGTAATCAGAGAGCCAGACAATGTGGACAATGCCAATG GCACCTGCTATGCCCTGATGAGCAGCACCACAACCACAACCACCACTCCTGATGGCA CCACCACCAGTGAGGAGGAGGAG

[0405] Codop RaCI4 ATGAGTGCCTTCAACATCTTTGCCCTGGTGGTGGTGGTGTGTGCCCTGATGATCAAT DNA sequence GAGTGCTGCACCAGCCAGGAGCCCACCACCCCCCTGAAGGCTGCCAGCCAGTGCAG with signal CAATGTGAAGTGCAGGAGAAGGTTTGACCACCTGGGCAACAGTGTGACAGAGGGCT peptide GCCCCAGTGGCTGCCTGTGTGTGTACCAAGCCACAGGCTACAACCAGGAGGCCAAT GGCACCTGCTATGAGCTGATGAAGACCAGCACCACAACCACTACAGAGGGCACCCC AGCCCAG

[0406] Codop RaCI5 ATGAATGCTGTGATTGTGCTGTGTGTGACCATCAGTGCTGTGCTGATCCACCAGTGC DNA sequence TACAGCACAGCTGAGGCCACCCTGAGCATCAATGGAGGAGACATGTGCATTGAGAAG with signal ACCTGCAACAGAAGCATTGATGCTGCTGGCAAGAAGGTCATTGCTGGCTGCCCTGGA peptide GGCTGCCTGTGTGTGTTCAATGTCAGTGATGTGACCTACCCTGCCAATGGCACCTGC TACCAGCTGGCCACCACAACCACTAACAGACCTGGAGCTGTGATGGAGAGAGAGAG G

[0407] Codop RaCI7 ATGGCAGCCCTGAATGGCCTGGTGCTGCTGCTGCTGACCATCAGTGCCATGTTCATA DNA sequence TCAGAGTGCTACAGTAGTGGAGAGAGCCAGAGCATCCAGAGGAAGGGCCAGTGTGA with signal GGAGGTGACCTGCCACAGAACCCTGAACCACCTGGGAGTGGCAGTGACCAGTGGCT peptide GCCCCAGTGGCTGCCTGTGTGTGATCAGTGCCCCAGACAGTGCAGTGAATGTGAAT GGCACCTGCTACCAGCTGATGGGCAGCACCAGCACAACCACTAGCAGCACCCCCAG CAGTGAGGACCAGGAG

[0408] Codop Omcl DNA ATGCTGGTGCTGGTGACCCTGATCTTCAGCTTCAGTGCCAACATTGCCTATGCAGAC sequence with AGTGAGAGTGACTGCACAGGCAGTGAGCCTGTGGATGCCTTCCAAGCCTTCAGTGA signal peptide GGGCAAGGAGGCCTATGTGCTGGTCAGAAGCACAGACCCCAAGGCCAGAGACTGCC TGAAGGGAGAGCCAGCAGGAGAGAAGCAGGACAACACCCTGCCAGTGATGATGACC TTCAAGAATGGCACAGACTGGGCCAGCACAGACTGGACCTTCACCCTGGATGGAGC CAAGGTCACAGCCACCCTGGGCAACCTGACCCAGAACAGAGAGGTGGTGTATGACA GCCAGAGCCACCACTGCCATGTGGACAAGGTGGAGAAGGAGGTGCCTGACTATGAG ATGTGGATGCTGGATGCTGGAGGCCTGGAGGTGGAGGTGGAGTGCTGCAGACAGAA GCTGGAGGAGCTGGCCAGTGGCAGAAACCAGATGTACCCCCACCTGAAGGACTGC

[0409]

[0410] CirpT consensus X1X2X3EX4GHX5YX6TX7NVTVEX8GACX9X10X11RNX12X13PX14GETKALX15X16PCVX17X18T without signal CYAAX19RX20VNX21TLCX22NX23GVX24X25GCX26X27X28WX29X30X31GX32YPX33CCPX34X35V peptide CX36X37X38X39X40X41X42

[0411] CirpT consensus X1X2X3EX4GHX5YX6X43X7NVTX44EX8GACX9X10X11RNX12X13PX14GETKALX15X16PCVX17X1 without signal 8TCYAX45X19RX20VNX21TLCX22NX23GVX24X25GCX26X27X28WX29X30X31GX32YPX33CCPX34 peptide X35VCX36X37X38X39X40X41X42

[0412] RaCI consensus X1CX2X3X4X5CX6RX7X8X9X10X11GX12X13X14X15X16X17CPX18GCLCVX19X20X21X22X23X24X25X without signal 26X27X28X29GTCX30X31L

[0413] peptide

[0414] RaCI consensus QCX1X2X3X4CX5RX6X7X8X9X10GX11X12X13X14X15X16CPX17GCLCVX18X19X20X21X22X23X24X25 without signal X26X27X28GTCX29X30L

[0415] peptide

[0416] Truncated 5’ ITR CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGA CCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAA CTCCATCACTAGGGGTTCCT

[0417] Truncated 3’ ITR AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTG AGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGT GAGCGAGCGAGCGCGCAG AAV 5 capsid MSFVDHPPDWLEEVGEGLREFLGLEAGPPKPKPNQQHQDQARGLVLPGYNYLGPGNGL DRGEPVNRADEVAREHDISYNEQLEAGDNPYLKYNHADAEFQEKLADDTSFGGNLGKAV FQAKKRVLEPFGLVEEGAKTAPTGKRIDDHFPKRKKARTEEDSKPSTSSDAEAGPSGSQ QLQIPAQPASSLGADTMSAGGGGPLGDNNQGADGVGNASGDWHCDSTWMGDRVVTK STRTWVLPSYNNHQYREIKSGSVDGSNANAYFGYSTPWGYFDFNRFHSHWSPRDWQR LINNYWGFRPRSLRVKIFNIQVKEVTVQDSTTTIANNLTSTVQVFTDDDYQLPYVVGNGTE GCLPAFPPQVFTLPQYGYATLNRDNTENPTERSSFFCLEYFPSKMLRTGNNFEFTYNFEE VPFHSSFAPSQNLFKLANPLVDQYLYRFVSTNNTGGVQFNKNLAGRYANTYKNWFPGP MGRTQGWNLGSGVNRASVSAFATTNRMELEGASYQVPPQPNGMTNNLQGSNTYALEN TMIFNSQPANPGTTATYLEGNMLITSESETQPVNRVAYNVGGQMATNNQSSTTAPATGT YNLQEIVPGSVWMERDVYLQGPIWAKIPETGAHFHPSPAMGGFGLKHPPPMMLIKNTPV PGNITSFSDVPVSSFITQYSTGQVTVEMEWELKKENSKRWNPEIQYTNNYNDPQFVDFA PDSTGEYRTTRPIGTRYLTRPL AAV5-derived MSFVDHPPDW LEEVGEGLRE FLGLEAGPPK PKPNQQHQDQ ARGLVLPGYN YLGPG capsid NGLDR

[0418] GEPVNRADEV AREHDISYNE QLEAGDNPYL KYNHADAEFQ EKLADDTSFG GNLGKAV FQA KKRVLEPFGL VEEGAKTAPT GKRIDDHFPK RKKARTEEDS KPSTSSDAEA GPSGSQQ LQI PAQPASSLGA DTMSAGGGGP LGDNNQGADG VGNASGDWHC DSTWMGDRVV TKST RTWVLP SYNNHQYREI KSGSVDGSNA NAYFGYSTPW GYFDFNRFHS HWSPRDWQRL INNYW GFRPR SLRVKI FNIQ VKEVTVQDST TTIANNLTST VQVFTDDDYQ LPYVVGNGTE GCLPAFPPQ V FTLPQYGYAT LNRDNTENPT ERSSFFCLEY FPSKMLRTGN NFEFTYNFEE VPFHSSFA PS QNLFKLANPL VDQYLYRFVS TNNTGGVQFN KNLAGRYANT YKNWFPGPMG RTQGW NLGSG VNRASVSAFA TTNRMELEGA SYQVPPQPNG MTNNLQGSNT YALENTMIFN SQPANP GTTA TYLEGNMLIT SESETQPVNR VAYNVGGQML AHKFKSGDAP ATGTYNLQEI VPGSVWM ERD VYLQGPIWAK IPETGAHFHP SPAMGGFGLK HPPPMMLIKN TPVPGNITSF SDVPVSSFI T QYSTGQVTVE MEWELKKENS KRWNPEIQYT NNYNDPQFVD FAPDSTGEYR TTRPIGT RYL TRPL

[0419]

[0420] AAV5-derived MSFVDHPPDW LEEVGEGLRE FLGLEAGPPK PKPNQQHQDQ ARGLVLPGYN YLGPG capsid NGLDR

[0421] GEPVNRADEVAREHDISYNE QLEAGDNPYL KYNHADAEFQ EKLADDTSFG GNLGKAV FQA KKRVLEPFGL VEEGAKTAPT GKRIDDHFPK RKKARTEEDS KPSTSSDAEA GPSGSQQ LQI PAQPASSLGA DTMSAGGGGP LGDNNQGADG VGNASGDWHC DSTWMGDRVV TKST RTWVLP SYNNHQYREI KSGSVDGSNA NAYFGYSTPW GYFDFNRFHS HWSPRDWQRL INNYW GFRPR SLRVKI FNIQ VKEVTVQDST TTIANNLTST VQVFTDDDYQ LPYVVGNGTE GCLPAFPPQ V FTLPQYGYAT LNRDNTENPT ERSSFFCLEY FPSKMLRTGN NFEFTYNFEE VPFHSSFA PS QNLFKLANPL VDQYLYRFVS TNNTGGVQFN KNLAGRYANT YKNWFPGPMG RTQGW NLGSG VNRASVSAFA TTNRMELEGA SYQVPPQPNG MTNNLQGSNT YALENTMIFN SQPANP GTTA TYLEGNMLIT SESETQPVNR VAYNVGGQML AHKFKSGDAP ATGTYNLQEI VPGSVWM ERD VYLQGPIWAK IPETGAHFHP SPAMGGFGLK HPPPMMLIKN TPVPGNITSF SDVPVSSFI T QYSTGQVTVE MEWELKKENS KRWNPEIQYT NNYNDPQFVD FAPDSTGEYR TTRPIGT RYL TRPL AAV25’ ITR TTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGT CGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGA GAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT AAV23’ ITR AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTG AGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGT GAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAA

[0422] OmCI variant DSESDCTGSEPVDAFQAWSEGKEAYVLVRSTDPKARDCLKGEPAGEKQDNTLPVMMTF without signal KNGTDWASTDWTFTLDGAKVTATLGNLTQNREVVYDSQSHHCHVDKVEKEVPDYEMW peptide MLDAGGLEVEVECCRQKLEELASGRNQMYPHLKDC

[0423] OmCI variant DSESDCTGSEPVDAFQAFSEGKEAYVLVRSTDPKARDCLKWEPAGEKQDNTLPVMMTF without signal KNGTDWASTDWTFTLDGAKVTATLGNLTQNREVVYDSQSHHCHVDKVEKEVPDYEMW peptide MLDAGGLEVEVECCRQKLEELASGRNQMYPHLKDC

[0424] OmCI variant DSESDCTGSEPVDAFQAFSEGKEAYVLVRSTDPKARDCLKGEPAGEKQDNTLPVMMTF without signal KNGTDWASTDWTFTLDGAKVTATLGNLTQNREVVYDSQSHHCHVDKVEKEVPDYEQW peptide QSNGSADDKEVECCRQKLEELASGRNQMYPHLKDC

[0425] OmCI variant DSESDCTGSEPVDAFQAFSEGKEAYVLVRSTDPKARDCLKGEPNGEKQDNTLPVMMTF without signal KNGTDWASTDWTFTLDGAKVTATLGNLTQNREVVYDSQSHHCHVDKVEKEVPDYEMW peptide QSDAGADAVEVECCRQKLEELASGRNQMYPHLKGC

[0426] OmCI variant DSESDCTGSEPVDAFQAFSEGKEAYVLVRSTDPKARDCLKGEPNGEKQDNTLPVMMTF without signal KNGTDWASTDWTFTLDGAKVTATLGNLTQNREVVYDSQSHHCHVDKVEKEVPDYEMW peptide QLDAGGDEVEVECCRQKLEELASGRNQMYPHLKGC

[0427] OmCI variant DSESDCTGSEPVDAFQAFSEGKEAYVLVRSTDPKARDCLKGEPNGEKQDNTLPVMMTF without signal KNGTDWASTDWTFTLDGAKVTATLGNLTQNREVVYDSQSHHCHVDKVEKEVPDYEMW peptide MLDAGGLEVEVECCRQKLEELASGRNQMYPHLKDC

[0428] OmCI variant DSESDCTGSEPVDAFQAFSEGKEAYVLVRSTDPKARDCLKGEPAGEKQDNTLPVMMTF without signal KQGTDWASTDWTFTLDGAKVTATLGQLTQNREVVYDSQSHHCHVDKVEKEVPDYEMW peptide MLDAGGLEVEVECCRQKLEELASGRNQMYPHLKDC

[0429] H1 protein QDLPGNDNSTATLCLGHHAVPNGTLVKTITNDQIEVTNATELVQSSSTGKICNNPHRILDGI sequence NCTLIDALLGDPHCDVFQDETWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFITE

[0430] GFTWTGVTQNGGSNACKRGPDSGFFSRLNWLTKSGSTYPVLNVTMPNNDNFDKLYIWG

[0431]

[0432] VHHPSTNQEQTSLYVQASGRVTVSTRRSQQTIIPNIGSRPWVRGLSSRISIYWTIVKPGDVLVINSNGNLIAPRGYFKMRTGKSSIMRSDAPIDTCISECITPNGSIPNDKPFQNVNKITYGA CPKYVKQNTLKLATGMRNVPEKQT

[0433] Codop AHSG ATGAAGTCCCTGGTCCTGCTCCTTTGTCTTGCTCAACTCTGGGGCTGCCACTCA sequence

[0434] Native CirpT4 ATGAAGTCCCTGGTCCTGCTCCTTTGTCTTGCTCAACTCTGGGGCTGCCACTCAGAC sequence with ATCCAGGAGCACGGCCACTCGTACTTGACGAGGAATGTGACTGTCGAGAATGGTGC AHSG signal CTGCATCTTTGAGCGTAACACGCTTCCTGATGGTGAAACCAAGGCACTTCATGATCCC peptide TGTGTCATCGCCACCTGCTACGCTGCGAGGCGCGAAGTGAACGCCACCCTGTGCCG

[0435] AAACTTCGGTGTCGACCCGGGCTGCAGGTTTCACTGGAGAAATGATGGTGTCTATCC

[0436]

[0437] TCAGTGCTGCCCAACACAAGTCTGCGATGGCACGGACTGAEMBODIMENTS OF THE INVENTION

[0438] 1. A vector for use in a method of treating an ocular disorder, wherein the vector comprises a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor and wherein the method comprises a step of intravitreal administration of the vector.

[0439] 2. A method of treating an ocular disorder comprising a step of intravitreal administration of a vector comprising a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor.

[0440] 3. Use of a vector in the manufacture of a medicament for use in a method of treating an ocular disorder, wherein the method comprises a step of intravitreal administration of the vector, wherein the vector comprises a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor.

[0441] 4. The vector for use, the method of treatment, or the use according to any one of the preceding claims, wherein the C5 inhibitor can bind to C5, and / or prevent cleavage of C5 to C5b by C5 convertase.

[0442] 5. The vector for use, the method of treatment, or the use according to any one of the preceding claims,, wherein the C5 inhibitor can bind to C5 with an affinity (Kd) of less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than InM.

[0443] 6. The vector for use, the method of treatment, or the use according to embodiment 5, wherein the affinity is measured by surface plasmon resonance (SPR).

[0444] 7. The vector for use, the method of treatment, or the use according to any one of embodiments 4 to 6, wherein preventing cleavage of C5 to C5b by C5 convertase is measured by a complement activity assay.

[0445] 8. The vector for use, the method of treatment, or the use according to embodiment 7, wherein the complement activity assay comprises incubating a solution comprising a known concentration of the C5 inhibitor and human serum, measuring the concentration of terminal complement complex (TCC) formed, for example using an antibody specific to the TCC, and calculating the level of reduction in TCC formation compared to an equivalent solution not comprising the C5 inhibitor.

[0446] 9. The vector for use, the method of treatment, or the use according to embodiment 7 or embodiment 8, wherein the C5 inhibitor can reduce TCC formation by at least 50%, or at least 70% of the level of reduction shown by a C5 inhibitor consisting of the amino acid sequence of SEQ ID NO: 5 in an equivalent complement activity assay.

[0447] 10. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor is a C5 inhibitor that can bind to the peripheral macro globulin domain 4 (C5 MG4) and / or peripheral macro globulin domain 5 (C5 MG5) of C5.11. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor is a polypeptide from the CirpT family. 12. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of any one of SEQ ID NOs: 6, 7, 84 or 85.

[0448] 13. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor is a polypeptide selected from the group consisting of a CirpT 1 polypeptide, a CirpT2 polypeptide, a CirpT3 polypeptide, and a CirpT4 polypeptide.

[0449] 14. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor is a CirpT4 polypeptide.

[0450] 15. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is:

[0451] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 50, at least 60, at least 70, or at least 80 amino acids of any one of SEQ ID NOs: 1-5;

[0452] (b) at least 95% identical to a fragment of at least 80 amino acids of any one of SEQ ID NOs: 1-5;

[0453] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 1- 5; or

[0454] (d) identical to any one of SEQ ID NOs: 1-5.

[0455] 16. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to a fragment of at least 80 amino acids of SEQ ID NO: 5.

[0456] 17. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to SEQ ID NO: 5.

[0457] 18. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 98% identical to SEQ ID NO: 5.

[0458] 19. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is:

[0459] (a) identical to SEQ ID NO: 5, but for one or more conservative amino acid substitutions at one or more positions corresponding to positions 1, 2, 3, 5, 8, 12, 18, 22, 23, 28, 30, 37, 42, 43, 51, 54, 60, 67, 78, 83 and 88 of SEQ ID NO: 5; and / or (b) identical to SEQ ID NO: 5, but for one or more conservative amino acid substitutions corresponding to one or more of DIE, I2V, Q3R, R5H, S8T, R12K,N18D, 122 V, F23Y, L28I, D30N, H37N, I42L, A43S, E51K, A54S, F60I, R67H, Q78N, Q83H, and T88A.

[0460] 20. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is identical to SEQ ID NO: 5.

[0461] 21. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 21 and 44 to 46.

[0462] 22. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9, wherein the C5 inhibitor can bind to the MG1, MG2, and / or C5d domains of C5.

[0463] 23. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22, wherein the C5 inhibitor is from the RaCI family.

[0464] 24. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22 to 23, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence of any one of SEQ ID NOs: 12, 86 or 87.

[0465] 25. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22 to 24, wherein the C5 inhibitor is a polypeptide selected from the group consisting of a RaCIl polypeptide, a RaCI2 polypeptide, a RaCI3 polypeptide, a RaCI4 polypeptide, a RaCI5 polypeptide, and a RaCI7 polypeptide.

[0466] 26. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22 to 25, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is:

[0467] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 50, at least 60, at least 65, at least 70, or at least 75 amino acids of any one of SEQ ID NOs: 8-11, 36, or 37;

[0468] (b) at least 95% identical to a fragment of at least 70 amino acids of any one of SEQ ID NOs: 8-11, 36, or 37;

[0469] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 8- 11, 36, or 37; and / or

[0470] (d) identical to any one of SEQ ID NOs: 8-11, 36, or 37.

[0471] 27. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22 to 26, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to any one of SEQ ID NOs: 8-11, 36, or 37.

[0472] 28. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22 to 27, wherein the C5 inhibitor comprises a polypeptide comprisingan amino acid sequence which is at least 98% identical to any one of SEQ ID NOs: 8-11, 36, or 37.

[0473] 29. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22 to 28, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is identical to any one of SEQ ID NOs: 8-11, 36, or 37.

[0474] 30. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 22 to 29, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 47 to 52. 31. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9, wherein the C5 inhibitor can bind to C5 and displaces the C5 C345C domain.

[0475] 32. The vector for use, the method of treatment, or the use of according to any one of embodiments 1 to 9 or 31, wherein the C5 inhibitor is an OmCI polypeptide.

[0476] 33. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 31 to 32, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is:

[0477] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 100, at least 110, at least 120, or at least 130 amino acids of any one of SEQ ID NOs: 38 or 95 to 101;

[0478] (b) at least 95% identical to a fragment of at least 130 amino acids of any one of SEQ ID NOs: 38 or 95 to 101;

[0479] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 38 or 95 to 101; and / or

[0480] (d) identical to any one of SEQ ID NO: 38 or 95 to 101.

[0481] 34. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 31 to 33, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to a fragment of at least 130 amino acids of any one of SEQ ID NOs: 38 or 95 to 101.

[0482] 35. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 31 to 34, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to any one of SEQ ID NOs: 38 or 95 to 101.

[0483] 36. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 31 to 35, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 98% identical to any one of SEQ ID NOs: 38 or 95 to 101.

[0484] 37. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 31 to 36, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is identical to any one of SEQ ID NOs: 38 or 95 to 101.38. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 9 or 31 to 37, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 53.

[0485] 39. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the polynucleotide and / or the transgene is codon optimised.

[0486] 40. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the polynucleotide and / or the transgene comprises a reduced number of CpGs compared to a wild type nucleotide sequence, optionally wherein the polynucleotide and / or the transgene is CpG-free.

[0487] 41. The vector for use, the method of treatment, or the use according to embodiment 39 or embodiment 40, wherein the polynucleotide and / or the transgene promotes expression of the C5 inhibitor at a higher level than an equivalent polynucleotide and / or transgene that is not codon-optimised.

[0488] 42. The vector for use, the method of treatment, or the use according to any one of embodiments 39 to 41, wherein the polynucleotide and / or the transgene promotes expression of the C5 inhibitor at a higher level than a polynucleotide and / or transgene comprising an equivalent native transgene, optionally the native transgene of SEQ ID NO: 21.

[0489] 43. The vector for use, the method of treatment, or the use according to any one of embodiments 39 to 42, wherein the polynucleotide and / or the transgene promotes at least 70% of the level of expression of a polynucleotide and / or transgene comprising SEQ ID NO: 22.

[0490] 44. The vector for use, the method of treatment, or the use according to any one of embodiments 41 to 43, wherein the expression is measured by transfecting a HEK293T cell sample with a plasmid or an AAV vector comprising the polynucleotide and / or the transgene, harvesting the supernatant of the cell sample and measuring the level of the C5 inhibitor, for example using an antibody specific for the C5 inhibitor.

[0491] 45. The vector for use, the method of treatment, or the use according to any one of embodiments 39 to 44, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 54 to 63. 46. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 21, or 39 to 44, wherein the polynucleotide comprises a nucleotide sequence which is:

[0492] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 175, at least 200, at least 225, or at least 250 nucleotides of any one of SEQ ID NOs: 22- 26;

[0493] (b) at least 95% identical to a fragment of at least 250 nucleotides of any one of SEQ ID NOs: 22-26;

[0494] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 22-26; and / or

[0495] (d) identical to any one of SEQ ID NOs: 22-26.47. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 21, 39 to 44 or 46, wherein the polynucleotide comprises a nucleotide sequence which is:

[0496] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 175, at least 200, at least 225, or at least 250 nucleotides of any one of SEQ ID NOs: 22, 23, 25 or 26;

[0497] (b) at least 95% identical to a fragment of at least 250 nucleotides of any one of SEQ ID NOs: 22, 23, 25 or 26;

[0498] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 22, 23, 25 or 26; and / or

[0499] (d) identical to any one of SEQ ID NOs: 22, 23, 25 or 26.

[0500] 48. The vector for use, the method of treatment, or the use according to embodiment 47, wherein the polynucleotide comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 22.

[0501] 49. The vector for use, the method of treatment, or the use according to embodiment 47 or embodiment 48, wherein the polynucleotide comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 22.

[0502] 50. The vector for use, the method of treatment, or the use according to any one of embodiments 47 to 49, wherein the polynucleotide comprises a nucleotide sequence which is identical to SEQ ID NO: 22.

[0503] 51. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor comprises a signal peptide, optionally the inhibitor’s native signal peptide.

[0504] 52. The vector for use, the method of treatment, or the use according to embodiment 51, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 64 to 66 or 70 to 76.

[0505] 53. The vector for use, the method of treatment, or the use according to embodiment 51, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 67 to 69 or 77 to 83.

[0506] 54. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 21, 39 to 44, or 46 to 51, wherein the polynucleotide comprises a nucleotide sequence which is:

[0507] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 200, at least 225, at least 250, at least 275, or at least 300 nucleotides of any one of SEQ ID NOs: 14-19;

[0508] (b) at least 95% identical to a fragment of at least 300 nucleotides of any one of SEQ ID NOs: 14-19;

[0509] (c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 14-19; and / or

[0510] (d) identical to any one of SEQ ID NOs: 14-19.55. The vector for use, the method of treatment, or the use according to embodiment 54, wherein the polynucleotide comprises a nucleotide sequence which is at least 95% identical to a fragment of at least 300 nucleotides of SEQ ID NO: 15.

[0511] 56. The vector for use, the method of treatment, or the use according to embodiment 54 or embodiment 55, wherein the polynucleotide comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 15.

[0512] 57. The vector for use, the method of treatment, or the use according to any one of embodiments 54 to 56, wherein the polynucleotide comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 15.

[0513] 58. The vector for use, the method of treatment, or the use according to any one of embodiments 54 to 57, wherein the polynucleotide comprises a nucleotide sequence which is identical to SEQ ID NO: 15.

[0514] 59. The vector for use, the method of treatment, or the use according to embodiment 51, wherein the signal peptide is a human protein-derived signal peptide, optionally an alpha-2-HS-glycoprotein / fetuin-A (AHSG) signal peptide.

[0515] 60. The vector for use, the method of treatment, or the use according to embodiment 51 or embodiment 59, wherein the signal peptide comprises an amino acid sequence:

[0516] (a) at least 90%, or at least 95% identical to SEQ ID NO: 13;

[0517] (b) identical to SEQ ID NO: 13 but for one, two, or three amino acid substitutions; and / or

[0518] (c) identical to SEQ ID NO: 13.

[0519] 61. The vector for use, the method of treatment, or the use according to any one of embodiments 51, 59 or 60, wherein the polynucleotide comprises a nucleotide sequence which is:

[0520] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 200, at least 225, at least 250, at least 275, or at least 300 nucleotides of SEQ ID NO: 20; (b) at least 95% identical to a fragment of at least 300 nucleotides of SEQ ID NO: 20;

[0521] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 20; and / or (d) identical to SEQ ID NO: 20.

[0522] 62. The vector for use, the method of treatment, or the use according to any one of embodiments 51 or 59 to 61, wherein the polynucleotide comprises a nucleotide sequence which is at least 95% identical to a fragment of at least 300 nucleotides of SEQ ID NO: 20.

[0523] 63. The vector for use, the method of treatment, or the use according to any one of embodiments 51 or 59 to 62, wherein the polynucleotide comprises a nucleotide sequence which is at least 95% identical to SEQ ID NO: 20.64. The vector for use, the method of treatment, or the use according to any one of embodiments 51 or 59 to 63, wherein the polynucleotide comprises a nucleotide sequence which is at least 98% identical to SEQ ID NO: 20.

[0524] 65. The vector for use, the method of treatment, or the use according to any one of embodiments 51 or 59 to 64, wherein the polynucleotide comprises a nucleotide sequence which is identical to SEQ ID NO: 20.

[0525] 66. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the vector is a viral particle, or a non-viral particle, optionally wherein the non-viral particle is a polynucleotide such as a plasmid or a nucleic acid minicircle, optionally formulated as a liposome, lipid nanoparticle, cationic polymer, microvesicle or exosome.

[0526] 67. The vector for use, the method of treatment, or the use according to embodiment 66, wherein the viral particle is an AAV particle, an adenovirus particle, a lentivirus particle, a retrovirus particle, or a herpes simplex virus particle.

[0527] 68. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the vector is an AAV particle comprising a capsid.

[0528] 69. The vector for use, the method of treatment, or the use according to embodiment 68, wherein the capsid is selected from the group consisting of an AAV1 capsid, an AAV2 capsid, an AAV3 capsid, an AAV4 capsid, an AAV5 capsid, an AAV6 capsid, an AAV7 capsid, an AAV8 capsid, an AAV9 capsid, an AAV10 capsid, an AAV11 capsid, or an AAV-derived capsid.

[0529] 70. The vector for use, the method of treatment, or the use according to embodiment 68 or embodiment 69, wherein the capsid is:

[0530] (a) an AAV2 capsid;

[0531] (b) an AAV2-derived capsid;

[0532] (c) an AAV6 capsid; or

[0533] (d) an AAV6-derived capsid.

[0534] 71. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of SEQ ID NO: 27-30.

[0535] 72. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 71, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of SEQ ID NO: 27-30.

[0536] 73. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 72, wherein the capsid comprises a polypeptide comprising an amino acidsequence at least 98% identical to a fragment of at least 700 amino acids of SEQ ID NO: 27- 30.

[0537] 74. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 73, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO: 27-30.

[0538] 75. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 74, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to SEQ ID NO: 27-30.

[0539] 76. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 75, wherein the capsid comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 27-30.

[0540] 77. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of SEQ ID NO: 31.

[0541] 78. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or embodiment 77, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of SEQ ID NO: 31.

[0542] 79. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70, 77 or 78, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of SEQ ID NO: 31.

[0543] 80. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 77 to 79, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO: 31.

[0544] 81. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 77 to 80, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to SEQ ID NO: 31.

[0545] 82. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 77 to 81, wherein the capsid comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 31.

[0546] 83. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.84. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 83, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

[0547] 85. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70, 83 or 84, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

[0548] 86. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 83 to 85, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

[0549] 87. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 83 to 86, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

[0550] 88. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 83 to 87, wherein the capsid comprises a polypeptide comprising an amino acid sequence of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

[0551] 89. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of SEQ ID NO: 35.

[0552] 90. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 89, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to a fragment of at least 650 amino acids of SEQ ID NO: 35.

[0553] 91. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70, 89 or 90, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to a fragment of at least 700 amino acids of SEQ ID NO: 35.

[0554] 92. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 89 to 91, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO: 35.

[0555] 93. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 89 to 92, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 98% identical to SEQ ID NO: 35.94. The vector for use, the method of treatment, or the use according to any one of embodiments 68 to 70 or 89 to 93, wherein the capsid comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 35.

[0556] 95. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the polynucleotide further comprises a transcription regulatory element.

[0557] 96. The vector for use, the method of treatment, or the use according to embodiment 95, wherein the transgene is operably linked to the transcription regulatory element.

[0558] 97. The vector for use, the method of treatment, or the use according to embodiment 95 or embodiment 96, wherein the transcription regulatory element comprises at least one element selected from the group consisting of a promoter, an enhancer, a post-transcriptional regulatory element, an intron, a 3’ UTR, and a 5’ UTR.

[0559] 98. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the polynucleotide comprises a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE).

[0560] 99. The vector for use, the method of treatment, or the use according to embodiment 98, wherein the WPRE comprises a polynucleotide comprising a nucleotide sequence which is:

[0561] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, or at least 550 nucleotides of SEQ ID NO: 40;

[0562] (b) at least 95% identical to a fragment of at least 550 nucleotides of SEQ ID NO: 40;

[0563] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 40; or (d) identical to SEQ ID NO: 40.

[0564] 100. The vector for use, the method of treatment, or the use according to embodiment 96 or embodiment 99, wherein the WPRE comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 550 nucleotides of SEQ ID NO: 40.

[0565] 101. The vector for use, the method of treatment, or the use according to any one of embodiments 98 to 100, wherein the WPRE comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 40.

[0566] 102. The vector for use, the method of treatment, or the use according to any one of embodiments 98 to 101, wherein the WPRE comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 40.

[0567] 103. The vector for use, the method of treatment, or the use according to any one of embodiments 98 to 102, wherein the WPRE comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 40.104. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the polynucleotide comprises a CBA promoter.

[0568] 105. The vector for use, the method of treatment, or the use according to embodiment 104, wherein the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is:

[0569] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, or at least 800 nucleotides of SEQ ID NO: 39;

[0570] (b) at least 95% identical to a fragment of at least 800 nucleotides of SEQ ID NO: 39;

[0571] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 39; or (d) identical to SEQ ID NO: 39.

[0572] 106. The vector for use, the method of treatment, or the use according to embodiment 104 or embodiment 105, wherein the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 800 nucleotides of SEQ ID NO: 39.

[0573] 107. The vector for use, the method of treatment, or the use according to any one of embodiments 104 to 106, wherein the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 39.

[0574] 108. The vector for use, the method of treatment, or the use according to any one of embodiments 104 to 107, wherein the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 39.

[0575] 109. The vector for use, the method of treatment, or the use according to any one of embodiments 104 to 108, wherein the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 39.

[0576] 110. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the recombinant genome further comprises 1 or 2 ITR sequences.

[0577] 111. The vector for use, the method of treatment, or the use according to embodiment 110, wherein the ITR sequences are AAV ITRs, optionally AAV2 ITRs.

[0578] 112. The vector for use, the method of treatment, or the use according to embodiment 110 or 111, wherein the ITR sequences comprise a 5’ ITR and a 3’ ITR.

[0579] 113. The vector for use, the method of treatment, or the use according to embodiment 112, wherein the 5’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 88 or 93.

[0580] 114. The vector for use, the method of treatment, or the use according to embodiment 112 or 113, wherein the 3’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 89 or 94.115. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the recombinant genome further comprises a polyA sequence.

[0581] 116. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the recombinant genome comprises at least one, at least two, or at least three polyA sequences.

[0582] 117. The vector for use, the method of treatment, or the use according to embodiment 115 or 116, wherein the polyA sequences are selected from a bGH polyA, a hGH polyA, or an SV40 polyA.

[0583] 118. The vector for use, the method of treatment, or the use according to embodiment 117, wherein the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is:

[0584] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 100, at least 125, at least 150, at least 175, or at least 200 nucleotides of SEQ ID NO: 41; (b) at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 41;

[0585] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 41; or (d) identical to SEQ ID NO: 41.

[0586] 119. The vector for use, the method of treatment, or the use according to embodiment 115 or embodiment 118, wherein the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 41.

[0587] 120. The vector for use, the method of treatment, or the use according to any one of embodiments 117 to 119, wherein the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 41.

[0588] 121. The vector for use, the method of treatment, or the use according to any one of embodiments 117 to 120, wherein the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 41.

[0589] 122. The vector for use, the method of treatment, or the use according to any one of embodiments 117 to 121, wherein the bGH polyA comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 41.

[0590] 123. The vector for use, the method of treatment, or the use according to embodiment 117, wherein the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is:

[0591] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 100, at least 125, at least 150, at least 175, or at least 200 nucleotides of SEQ ID NO: 42; (b) at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 42;

[0592] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 42; or(d) identical to SEQ ID NO: 42.

[0593] 124. The vector for use, the method of treatment, or the use according to embodiment 117 or embodiment 123, wherein the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 200 nucleotides of SEQ ID NO: 42.

[0594] 125. The vector for use, the method of treatment, or the use according to any one of embodiments 117, 123 or 124, wherein the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 42.

[0595] 126. The vector for use, the method of treatment, or the use according to any one of embodiments 117 or 123 to 125, wherein the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 42.

[0596] 127. The vector for use, the method of treatment, or the use according to any one of embodiments 117 or 123 to 126, wherein the hGH polyA comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 42.

[0597] 128. The vector for use, the method of treatment, or the use according to embodiment 117, wherein the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is:

[0598] (a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 100, at least 125, at least 150, or at least 175 nucleotides of SEQ ID NO: 43;

[0599] (b) at least 95% identical to a fragment of at least 175 nucleotides of SEQ ID NO: 43;

[0600] (c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 43; or (d) identical to SEQ ID NO: 43.

[0601] 129. The vector for use, the method of treatment, or the use according to embodiment 115 or embodiment 128, wherein the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to a fragment of at least 175 nucleotides of SEQ ID NO: 43.

[0602] 130. The vector for use, the method of treatment, or the use according to any one of embodiments 117, 128, or 129, wherein the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 95% identical to SEQ ID NO: 43.

[0603] 131. The vector for use, the method of treatment, or the use according to any one of embodiments 117 or 128 to 130, wherein the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is at least 98% identical to SEQ ID NO: 43.

[0604] 132. The vector for use, the method of treatment, or the use according to any one of embodiments 117 or 128 to 131, wherein the SV40 polyA comprises a polynucleotide comprising a nucleotide sequence which is identical to SEQ ID NO: 43.133. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 3, wherein the vector comprises, in 5’ to 3’ order, a 5’ ITR, a promoter, a transgene encoding a C5 inhibitor, a WPRE sequence, a polyA sequence, and a 3’ ITR.

[0605] 134. The vector for use, the method of treatment, or the use according to any one of embodiments 1 to 3, wherein the vector comprises, in 5’ to 3’ order, a 5’ ITR, a promoter, a transgene encoding a C5 inhibitor, a WPRE sequence, three polyA sequences, and a 3’ ITR.

[0606] 135. The vector for use, the method of treatment, or the use of according to embodiment 133 or embodiment 134, wherein the 5’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 88 or 92.

[0607] 136. The vector for use, the method of treatment, or the use of according to any one of embodiments 133 to 135, wherein the promoter comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 39.

[0608] 137. The vector for use, the method of treatment, or the use of according to any one of embodiments 133 to 136, wherein the transgene comprises a nucleotide sequence which is at least 98% identical or identical to SEQ ID NO: 15 or SEQ ID NO: 20.

[0609] 138. The vector for use, the method of treatment, or the use of according to any one of embodiments 133 to 137, wherein the WPRE sequence comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 40.

[0610] 139. The vector for use, the method of treatment, or the use of according to any one of embodiments 133 and 135 to 138, wherein the polyA sequence comprises a nucleotide sequence which is at least 95% identical, 98% identical, or identical to SEQ ID NO: 41. 140. The vector for use, the method of treatment, or the use of according to any one of embodiments 134 to 138, wherein the three polyA sequences comprise a nucleotide sequence comprising (a) a sequence which is at least 95% identical, at least 98% identical or identical to SEQ ID NO: 41; (b) a sequence which is at least 95% identical, at least 98% identical or identical to SEQ ID NO: 42; and (c) a sequence which is at least 95% identical, at least 98% identical or identical to SEQ ID NO: 43.

[0611] 141. The vector for use, the method of treatment, or the use of according to any one of embodiments 133 to 140, wherein the 3’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical or identical to SEQ ID NO: 89 or 93.

[0612] 142. The vector for use, the method of treatment, or the use of according to any one of the preceding embodiments, wherein the vector can prevent cleavage of C5 to C5b by C5 convertase.

[0613] 143. The vector for use, the method of treatment, or the use of according to embodiment 142, wherein the vector has at least 70% of the C5 inhibitory activity of an equivalent vector comprising a promoter comprising the nucleotide sequence which is identical to SEQ ID NO: 39 and a transgene comprising a nucleotide sequence which is identical to SEQ ID NO: 21 in an equivalent complement activity assay.144. The vector for use, the method of treatment, or the use of according to embodiment 142 or embodiment 143, wherein preventing cleavage of C5 to C5b by C5 convertase and / or C5 inhibitory activity is measured by a complement activity assay.

[0614] 145. The vector for use, the method of treatment, or the use of according to embodiment 144, wherein the complement activity assay comprises transfecting a HEK293T cell sample with the vector, collecting the supernatant, preparing supernatant solutions by serial dilution, incubating a known volume of the supernatant solutions with human serum, measuring the concentration of terminal complement complex (TCC) formed, for example using an antibody specific to the TCC, and calculating the level of reduction in TCC formation compared to an equivalent solution comprising human serum and the supernatant of an equivalent untransfected HEK293T cell sample.

[0615] 146. The vector for use, the method of treatment, or the use of according to any one of the preceding embodiments wherein the method of treatment is a method of gene therapy.

[0616] 147. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the vector is part of a composition.

[0617] 148. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the ocular disorder is a complement-mediated disorder, a disorder associated with C5 and / or a condition that would benefit from reducing complement-mediated inflammation.

[0618] 149. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the ocular disorder is selected from age-related macular degeneration (AMD), geographic atrophy (GA), diabetic macular edema (DME), diabetic retinopathy, ocular angiogenesis (ocular neovascularization affecting choroidal, corneal or retinal tissue), uveitis, glaucoma, and neuromyelitis optica.

[0619] 150. The vector for use, the method of treatment, or the use according to embodiment 149, wherein the ocular disorder is age-related macular degeneration (AMD) or geographic atrophy (GA).

[0620] 151. The vector for use, the method of treatment, or the use according to embodiment 150, wherein the AMD is dry AMD, optionally wherein the AMD comprises GA.

[0621] 152. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the subject is heterozygous or homozygous for the R885C / H polymorphism of C5.

[0622] 153. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein intravitreal administration comprises administration of the polynucleotide, polypeptide, viral particle, or the composition to the vitreous humour using a needle.154. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the ocular disorder is a degenerative disorder, optionally wherein the affected cells may have an impaired ability, relative to unaffected cells, to be successfully transduced by a gene therapy vector and produce therapeutic protein.

[0623] 155. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the step of intravitreal administration of the vector is a step of intravitreal administration of the vector at a dose of less than 1 x 1012vg, less than 9 x 1011vg, less than 8 x 1011vg, less than 7 x 1011vg, less than 6 x 1011vg, less than 5 x 1011vg, less than 4 x 1011vg, less than 3 x 1011vg, less than 2 x 1011vg, less than 1 x 1011vg, less than 8 x IO10vg, or less than 6 x IO10vg per eye.

[0624] 156. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein C5 inhibitor is nor an antibody or a fragment or variant thereof.

[0625] 157. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitors consists of 200 amino acids or fewer. 158. The vector for use, the method of treatment, or the use according to any one of the preceding embodiments, wherein the C5 inhibitor is a non-mammalian C5 inhibitor.

[0626] 159. The vector for use, the composition for use, the method of treatment, the use, or the method of any one of the preceding embodiments, wherein the vector or composition is administered at a dose of 2.5 x IO10vg, 7.5 x IO10vg, or 2.5 x 1011vg / eye.

[0627] 160. The vector for use, the composition for use, the method of treatment, the use, or the method of any one of the preceding embodiments, wherein the vector or composition is administered at a dose of between 2.5 x IO10vg per eye and 2.5 x 1011vg per eye, between 2.5 x IO10vg per eye and 7.5 x IO10vg per eye or between 7.5 x 1010vg per eye and 2.5 x 1011vg per eye.EXAMPLES

[0628] Example 1 - In vitro evaluation of potential complement pathway inhibitors

[0629] To assess the potency of potential complement inhibitors an ELISA assay that centres on detection of the terminal complement complex (TCC; also called the membrane-attack complex (MAC)), in vitro in serum was used. In this assay, activation of the complement alternative pathway (AP) is triggered by lipopolysaccharide (LPS) which leads the deposition of the TCC. The TCC is then detected by a monoclonal anti-C5b-9 + C5b-8 antibody [aEl 1] (ab66768; Abeam) that binds the interfaces between C5b and both C8 & C9. The aEl 1 antibody does not react with native C9 or C8 proteins.

[0630] Transgenes encoding the potential inhibitor proteins were codon optimized. A His-tag was added to the C-terminal of the transgenes to allow for detection and quantification, and the respective native signal peptide coding sequence was replaced with the coding sequence (codon-optimised) for the human Alpha-2-HS-Glycoprotein (AHSG) signal peptide (SEQ ID NO: 103), with the aim of efficient protein secretion. The transgenes were synthesized and cloned into a common plasmid backbone under the control of a CMV promoter, and the nucleotide sequence was validated by sequencing.

[0631] To express the transgenes, 3 x 105HEK293T cells were seeded per well in 24-well plates in 600 pL DMEM (Gibco cat. # 11995073) supplemented with 10% FBS (Cytiva HyClone cat. #: SH30071.03HI). 24 hours after seeding, the cells were transfected with 600 ng plasmid DNA per well carrying the respective inhibitor transgenes using 1.8 pl FuGENE® HD transfection reagent (Promega Cat. #: E2311) in 47 pl Opti-MEM (Gibco, cat. #: 31-985-062) following manufacturer’s instructions and incubated at 37°C, 5% CO2. 24 hours posttransfection, the complete medium was removed and replaced with 600 pL pre-warmed serum free Opti-MEM medium. Cells were then incubated at 37°C, 5% CO2 for 72 hours. The culture conditioned medium (supernatant) was harvested, cleared by centrifugation at 2000 x g for 10 minutes at 4°C, and stored at -80°C for further characterization.

[0632] For the TCC formation inhibition assay, a 96-well microtiter plate was coated with lipopolysaccharide (LPS: O26:B6 Cat #. L3755-100MG) at llpg / mL (70 pL per well) overnight at 4°C. After the incubation, the plate was washed four times with wash buffer (PBS:Tween 0.05%). 60 pL undiluted supernatant from HEK293T transfection was mixed with 10 pL human serum complement-preserved gender-pooled from BioIVT (BIOIVT HLMANSRM-0103036) pre-diluted 1:1 with alternative pathway buffer (lOmM HEPES pH7.3, 140mM NaCl, lOmM EGTA, 2mM MgCl). 50 pl of the mixture was applied to the LPS coated plate and incubated for 3 hours at 37°C with agitation (150rpm). Recombinant human complement receptor 1 (CR1, CD35) was used as positive control as it inhibits C3 and C5 convertases of the three complement pathways, namely the classical, alternative, and lectin. At the end of the incubation, the plate was washed four times with wash buffer and 50 pl aEl 1 monoclonal antibody diluted to 2.5 pg / ml in ELISA General Assay Diluent buffer (Bio-Rad cat. #: BUF037C) was applied, followed by incubation for 1 hour at room temperature. The plate was then washed four times and 50 pl secondary Peroxidase AffiniPure™ Donkey Anti-Mouse IgG (H+L) antibody (Jackson Immuno Research 715-035-151) was applied at 1:10,000 dilution. After 1 hour incubation at room temperature, the plate was washed four times followed by addition of 50 pl of 1-Step™ TMB ELISA Substrate Solutions (Thermo Fisher cat. #: 34028) and incubated in the dark for 15 minutes. 50 pl Stopsolution (2 M H2SO4) was applied to the plate and the OD was measured at 450 nm using SpectraMax i3x plate reader (Molecular Devices) and Softmax pro 7 software.

[0633] As shown in Figure 1, the inhibitors showed strong inhibition of the TCC formation as reflected by low absorbance values compared to the GFP control supernatant used as negative control. The human recombinant CR1 protein (rCRl) used as positive control showed a strong inhibitory activity on the TCC formation as expected.

[0634] C5 Inhibitor SEQ ID NO

[0635] CirpTl 1

[0636] CirpT2 3

[0637] CirpT3 4

[0638] CirpT4 5

[0639] OmCI 38

[0640] RaCIl 8

[0641] RaCI2 9

[0642] RaCI3 10

[0643] RaCI4 11

[0644] RaCI5 36

[0645]

[0646] RaCI7 37

[0647] Table 5 - Amino acid sequences for each of the C5 inhibitors without signal peptide.

[0648] Example 2 - Enhancement of CirpT4 expression levels

[0649] A number of codon-optimized CirpT4 genes were synthesized (see Table 5), cloned into an AAV production plasmid under the control of a CBA promoter, and assessed for protein expression relative to the native CirpT4 coding sequence. With the aim of further enhancing CirpT4 expression levels, a Woodchuck hepatitis virus Posttranscriptional Regulatory Element (WPRE) was placed at the 3' end of the CirpT4 gene, followed by a bovine growth hormone (bGH) polyadenylation sequence (SEQ ID NO: 41).

[0650] Construct SEQ ID NO

[0651] NATIVEsp-CirpT4-wt 14

[0652] AHSGsp-CirpT4-wt 104

[0653] NATIVEsp-CirpT4-Co02 16

[0654] NATIVEsp-CirpT4-Co03 17

[0655] NATIVEsp-CirpT4-Co04 18

[0656] NATIVEsp-CirpT4-Co05 19

[0657]

[0658] AHSGsp-CirpT4-Col 20

[0659] Table 6 - Sequences for each of the transgenes including signal peptide and the gene, but not including WPRE.

[0660] Five codon-optimized CirpT4 genes (with or without a C-terminal His-tag) were generated, in each case including either the native CirpT4 signal peptide coding sequence (NATIVEsp) or instead carrying the heterologous human alpha-2-HS-glycoprotein signal peptide (AHSGsp) coding sequence. Plasmids were transformed into XLIO-Gold ultracompetent E. coli cells for amplification, and clones were selected for validation. Next generation sequencing of wholeplasmids, and restriction digests (with appropriate restriction sites to check for transgene insertion and ITR integrity), were performed to validate the codon-optimized plasmids.

[0661] First, the impact of the AHSGsp on the expression level, in supernatant of transfected HEK293T cells, of the native (i.e. non-codon-optimized) CirpT4-6xHis gene (“CirpT4-wt”) was analyzed by Western blot and ELISA assay. Briefly, 5 x 105HEK293T cells were seeded in 6-well plates in 2 ml DMEM medium (Gibco cat. #: 11995073) supplemented with 10% FBS (Cytiva HyClone cat. #: SH30071.03HI). 24 hours after seeding, the cells were transfected with 2 pg plasmid DNA encoding CirpT4-wt incorporating either the native or AHSG signal peptides, using ATCC GenXPlus transfection reagent (Cat. # ACS-4004) following manufacturer’s instructions. Negative controls consisted of untransfected cells. 24 hours post-transfection, the complete medium was removed, cells were washed with 1ml prewarmed serum-free Opti-MEM, and the medium was then replaced with 2 ml of pre-warmed Opti-MEM. Cells were incubated at 37°C, 5% CO2 for 72 hours. The culture medium (supernatant) was harvested, cleared by centrifugation at 2000 x g for 10 minutes at 4°C, ali quoted and stored at -80°C for further characterization.

[0662] CirpT4 protein concentrations were measured by a His-tag ELISA detection kit (Genscript Cat. #: L00436) following manufacturer’s instructions and using supernatant diluted 1 in 100 with ELISA Assay Diluent. For Western blot analysis, 37.5 pl cleared supernatant was mixed with 12.5 pl 4x SDS Laemmli buffer (Bio-Rad Cat #: 1610747) containing 2-Mercaptoethanol (Bio-Rad Cat# 1610710), incubated at 95°C for 10 min, resolved by SDS-PAGE and transferred to a PVDF membrane. Immunoblot analysis was performed with the indicated antibodies and visualized with SuperSignal West Femto Chemiluminescent substrate (Thermo Scientific Cat #: 34096) and images were taken using Bio-Rad ChemiDoc XRS+ with Image Lab software.

[0663] As shown in Figure 2, CirpT4 expression levels were comparable when using NATIVEsp or AHSGsp signal peptides. CirpT4 with the NATIVEsp produced a mean of 893 ng / ml and with the human AHSGsp a mean of 876 ng / ml CirpT4 protein (Figure 2B). Both CirpT4-wt proteins migrated at a molecular weight of 25 kDa as shown in the Western blot analysis (Figure 2A).

[0664] Next, the protein expression from codon-optimized CirpT4 genes (numbered “CoOl” to “Co05”) in supernatant of transfected HEK293T cells was analyzed by Western blot and a newly-generated CirpT4 polyclonal antibody (T00710) at 1: 1000 dilution. (The native “CirpT4-wt” and codon-optimised “CirpT4-Co..” genes used in this experiment, resulting in the data shown in Figure 3, did not encode a HIS-tag.) Western blot revealed that CirpT4-CoOl with AHSGsp and a WPRE element placed at the 3’ end of the CirpT4-Co01 coding sequence (AHSGsp-CirpT4-Co01_WPRE) resulted in an around 4-fold increase in expression compared to the native CirpT4 sequence. AHSGsp-CirpT4-Co01_WPRE produced the highest CirpT4 protein levels compared to other CirpT4 codon optimizations (Figure 3B). The use of WPRE enhanced CirpT4-Co01 expression compared to CirpT4-Co01 without WPRE. CirpT4 protein expressed from the codon-optimized genes migrated at a molecular weight of 25 kD on SDS-PAGE, comparable to CirpT4-wt (Figure 3 A).

[0665] Example 3 - In vitro Alternative Complement Pathway (AP) inhibition ELISA assay to assess the activity of codon-optimised CirpT4 genesHycult Human Alternative Complement Pathway ELISA Kit (HK3012) was used to assess and compare the activity of codon-optimized CirpT4 genes in alternative complement pathway inhibition. The HK3012 AP complement activity assay measures complement function in vitro in serum.

[0666] The wells of the assays are coated with lipopolysaccharide (LPS) and when the cascade has elapsed, the in vitro-formed terminal complement complex (TCC), also known as the MAC, is detected with a C9 neo-epitope antibody. The TCC contains interfaces between C5b and both C8 & C9. These interfaces are detectable via the C9 neo-epitope antibody. The assay was performed using supernatant of HEK293T cells transfected with plasmids carrying respective CirpT4 codon-optimized, as well as native, coding sequences, and human serum complement-preserved gender-pooled from BioIVT (BIOIVT HUMANSRM-0103036). In the assay, 13.3 % human serum was used and mixed with same volume of supernatant that had been serially diluted (50 % to 2.9 %) in ELISA dilution buffer. 100 pl mixture was then loaded into the Hycult ELISA plate wells and the assay was performed following manufacturer’s instructions. The results shown in Figure 4 reveal that CirpT4 codon-optimization resulted in a greater inhibitory effect on the TCC formation compared to the native CirpT4 gene, due to the observed increase in protein expression levels (Figure 3). Increasing levels of CirpT4 in the assay lead to stronger inhibition of the TCC formation. AHSGsp-CirpT4-Co01_WPRE showed 95 % TCC inhibition at 9.9 % supernatant, followed by AHSGsp-CirpT4-Co01, which reached 93 % TCC inhibition at 14.8 % supernatant. AHSGsp-CirpT4-wt reached 91 % TCC inhibition at 50% supernatant. This data correlates with the increased expression levels of AHSGsp-CirpT4-Co01_WPRE and AHSGsp-CirpT4-Co01 as shown in the Figure 3 Western blot. Untransfected cell supernatant (Neg Ctrl) showed no inhibition.

[0667] Example 4 - Expression of CirpT4 in HEK293T cells following transduction with crude AAV-CirpT4

[0668] Small scale AAV production

[0669] Small scale AAV production by triple transfection of HEK293T cells was performed using TransIT-VirusGEN (Minis cat# MIR 6700). Briefly, 1 x 106HEK293T cells were seeded in 6-well plate in 2 ml DMEM supplemented with 10 % FBS. 24 hours post seeding, medium was replaced with DMEM supplemented with 2 % FBS. 3 pg total DNA comprising the ‘gene-of-interest’ plasmid pAAV-CirpT4, the AAV rep and cap gene plasmid pRep2Cap, and the pALD-X80 helper plasmid, with equal molar ratios 1:1:1, was added to 200 pL of Opti-MEM in a sterile tube. 6 pl TransIT-VirusGEN® Reagent was added to the Opti-Mem / DNA mixture. After 15 minutes incubation at room temperature, the mixture was added dropwise to the well and the plate incubated at 37°C, 5% CO2 for 72 hours. Cells and supernatant were collected and transferred to a 15 ml falcon. After three cycles of freeze on dry ice ethanol and thaw in 37°C water bath, the samples were treated with Benzonase endonuclease EMPROVE® EXPERT (Millipore Sigma cat. #: 101697) at a final concentration of 25 U / mL and incubated in 37°C water bath for 90 minutes with vortexing every 20 minutes. The samples were then spun down at 10,000 x g for 10 min at 4°C to remove cell debris, and cleared supernatant containing the AAV particles was aliquoted and stored at -80°C for further characterization. Titration of the AAV particles was performed using qPCR with ITR primers and probe.Expression of CirpT4 from crude AAV in HEK293T cells.

[0670] I x IO5HEK293T cells were seeded in a 24-well plate coated with poly-D-Lysine (Thermo scientific cat. # 152025) in 500 pl DMEM supplemented with 10% FBS. 24 hours post seeding, the cells were transduced with crude AAV from the small scale AAV production as described above at an MOI of 1 x 105in a total volume of 500 pl plain DMEM in the presence of Calpain Inhibitor I (Sigma cat. #: A6185) at 75 pM as transduction enhancer. 5 hours post transduction, the wells were topped with 500 pl DMEM supplemented with 10% FBS and the plate was incubated at 37°C, 5% CO2 for 72 hours. The conditioned supernatant was collected, centrifuged at 2000 x g for 10 min at 4°C to remove cell debris, and the cleared supernatant was aliquoted and stored at -80°C for further characterization.

[0671] Assessment of CirpT4 potency in HEK293T cells supernatant following crude AAV-CirpT4 transduction

[0672] Supernatants were used in the in vitro Hycult TCC inhibition ELISA assay as described above. AHSGsp-CirpT4-Co01_WPRE showed 90% inhibition of TCC formation at 14.8% supernatant in the assay, compared to AHSGsp-CirpT4-Co01 without WPRE which showed 41% TCC inhibition at the same supernatant concentration (Figure 5). The untransduced cell supernatant (Negative control) showed no inhibition.

[0673] Example 5 - Immunogenicity risk assessments of CirpT4: in silica prediction

[0674] As the first step to assess the immunogenicity risk of CirpT4 (SEQ ID NO: 5), the NetMHC4.0 server (https : / / services. healthtech. dtu.dk / services / NetMHC-4.0 / ; Reliable prediction of T-cell epitopes using neural networks with novel sequence representations, Nielsen M et al, Protein Sci., (2003) 12:1007-17; Gapped sequence alignment using artificial neural networks: application to the MHC class I system, Andreatta M and Nielsen M, Bioinformatics (2016) Feb 15;32(4): 511-7) was used to predict binding of peptides to MHC class I molecules, and NETMHCIIpan4.1 (https: / / services.healthtech.dtu.dk / services / NetMHCIIpan-4.1 / ; Improved prediction of MHC II antigen presentation through integration and motif deconvolution of mass spectrometry MHC eluted ligand data, Reynisson B et al, J Proteome Res 2020 Apr 30) to predict binding of peptides to MHC class II molecules.

[0675] The NetMHC4.0 server was used to screen inhibitors for 8-12mer HLA-A*02:01 binders with a threshold of 0.5 for strong binders and a threshold of 2.0 for weak binders.

[0676] NETMHCIIpan4.1 was used to find 15-22mer peptides which bind a range of HLA composed of chains from the HLA-DP locus. Hemagglutinin was used as the positive control. As shown in Figure 6A, CirpT4 showed very few HLA-I binding peptides with 5 weak binders and 1 strong binder compared to the Influenza hemagglutinin (HA) protein positive control with 29 weak and 7 strong binders. In addition, CirpT4 did not show any predicted peptide binders to HLA-II, compared to Hemagglutinin Hl positive control with 26 weak and 8 strong binders (Figure 6B).

[0677] Example 6 - B-cell reactivity to CirpT4

[0678] To assess CirpT4 protein pre-existing humoral immunity, human B-cell supernatant was screened for IgG reactive to CirpT4 (SEQ ID NO: 5). In this assay, clonal B-cell populations were differentiated into B cells that produce IgG. The B cells were isolated from PBMCs (BioIVT, HUMANPBMC-0002409) from normal donors using a Human B-Cell Isolation Kit II (Miltenyi Biotec cat. #: 130-091-151). The B-cells were then resuspended in Lymphocytesupplemented medium and 16,000 cells were applied per well in a final volume of 100 pL / well into a 384-well plate coated with CD40L (Biotechne cat. # 6420-CL) at Ipg / mL. After 14 days of incubation at 37°C, 5% CO2, 70 pl supernatant containing secreted IgG was harvested and the concentration of IgG was determined by ELISA-based assay.

[0679] Briefly, a 96-well microplate was coated with recombinant human Complement Factor H (Complement Technology cat. #: A137), Hemagglutinin 1 (Abeam cat. #: AB217651), or CirpT4 protein (Genescript C5IT4 AMBAM) at 0.03 pM in lx Coating Buffer (Bio-rad cat. # BUF030A) and incubated overnight at 4°C. The next day, the plate was washed three times with 250 pL wash buffer (PBS:Tween 0.05%), followed by a blocking step with 5.0% BSA for 1 hour at room temperature. The plate was then washed 3 times. 60 pl B cell supernatant was mixed with 190 pl assay diluent and 50 pl of the mixture was added to the wells, followed by incubation for 1 hour at 37°C with agitation (150rpm). After three washes, 50 pl horseradish peroxidase (HRP)-conjugated goat anti-human IgG (Jackson Immuno Research cat. # 109-035-003) at 1 : 10,000 dilution in assay diluent was added to the wells, followed by incubation for 1 hour at room temperature. The plate was washed three times followed by addition of 50 pl of 1-Step™ TMB ELISA Substrate Solutions (Thermo Fisher cat. # 34028) and incubation in the dark for 15 minutes. 50 pl Stop solution (2 M H2SO4) was applied to the plate and the OD was measured at 450 nm using SpectraMax i3x plate reader (Molecular Devices) using Softmax pro 7 software.

[0680] The results showed that 60% of B-cell populations from donor 1 and 28% of B-cell populations from donor 2 secrete IgG reactive to Influenza Hemagglutinin 1 protein, while a very low percentage of B-cell populations secrete IgG reactive to CirpT4 and CFH (Figure 7).

[0681] Example 7 - Intravenous immunoglobulin reactivity to CirpT4

[0682] Intravenous immunoglobulin (IVIg) is a mixture of IgG and other antibodies derived from thousands of healthy human plasmas.

[0683] To determine whether IVIg has detectable CirpT4 reactive antibodies, an IgG detection ELISA assay was developed. Briefly, a 96-well microplate was coated with recombinant human Complement Factor H (Complement Technology cat. #: A137), Hemagglutinin 1 (Abeam cat. # AB217651), or CirpT4 protein (SEQ ID NO: 5) (Genescript C5IT4_AMBAM) at a range of concentrations and incubated overnight at 4°C. The next day, the plate was washed three times with 250 pL wash buffer (PBS:Tween 0.05%), followed by a blocking step with 5 % BSA for 1 hour at room temperature. The plate was then washed 3 times. 50 pL IVIg (Creative Biomart Cat#: THP-0108) at 23.3 pg / ml assay diluent solution was applied per well and the plate was incubated for Ihour at 37°C. IVIg contains the same distribution of IgG antibody subclasses as is found in the general human population. IgG subclasses are fully represented in the following proportions: 70.3% IgGl, 24.7% IgG2, 3.1% IgG3, and 1.9% IgG4. After washing the plate four times with wash buffer, 50 pl horseradish peroxidase (HRP)-conjugated goat anti -human IgG (Jackson Immuno Research cat. #: 109-035-003) at 1 : 10,000 dilution in assay diluent was added to the wells, followed by incubation for 1 hour at room temperature. The plate was washed four times followed by addition of 50 pl of 1-Step™ TMB ELISA Substrate Solutions (Thermo Fisher cat. # 34028) and incubation in the dark for 15 minutes. 50 pl Stop solution (2M H2SO4) was applied to the plate and the OD was measured at 452 nm using SpectraMax i3x plate reader (Molecular Devices) using Softmax pro 7 software.The results of three tested IVIg lots were combined and showed that IVIg from thousands of healthy donors have no presence of pre-existing anti-CirpT4 antibodies as compared to the CFH negative control (Figure 8). In contrast, IgG binding of Hemagglutinin was detected in a dose dependent manner.

[0684] Example 8 - Optimising transgene cassette size

[0685] The transduction efficiency of two vectors (with and without additional hGH and SV40 polyA sequences) was assessed to ensure no decrease in CirpT4 expression was observed with the longer transgene cassette / vector genome. The sequences of the elements of the two vectors are provided in Table 7 below.

[0686] Sequence element SEQ ID NO

[0687] CBA promoter 39

[0688] CirpT4 transgene including AHSG signal 20

[0689] peptide

[0690] WPRE 40

[0691] bGH polyA 41

[0692] hGH polyA 42

[0693]

[0694] SV40 polyA 43

[0695] Table 7: Vector element sequences

[0696] 1 x 105HEK293T cells were seeded in a 24-well plate coated with poly-D-Lysine (Thermo scientific cat. # 152025) in 500 pl DMEM supplemented with 10% FBS. 24 hours post seeding, the cells were transduced with crude AAV from the small scale AAV production as described above at an MOI of 1 x 105in a total volume of 500 pl plain DMEM in the presence of Calpain Inhibitor I (Sigma cat. #: A6185) at 75 pM as transduction enhancer. 5 hours post transduction, the wells were topped with 500 pl DMEM supplemented with 10% FBS and the plate was incubated at 37°C, 5% CO2 for 72 hours. The conditioned supernatant was collected, centrifuged at 2000 x g for 10 min at 4°C to remove cell debris, and the cleared supernatant was aliquoted and stored at -80°C for further characterization.

[0697] It was found that the longer vector genome did not negatively impact in vitro CirpT4 expression and even led to a modest increase in protein expression (Figure 10(A)).

[0698] Expanding the transgene beyond half of the length of the wild type AAV genome (approximately 4.7 kb) by adding extra polyA sequences prevented dimerization from occurring. Although CB A-CirpT4-WPRE-bGH expressed well, due to the relatively small size of this vector genome (~2.3 kb ITR-ITR) dimerization was frequently observed when vector quality was assessed by alkaline gel electrophoresis. Expanding the vector genome to 2.8 kb by adding non-homologous polyA sequences -well beyond the 2.35 kb threshold (half of the length of the wild type AAV genome) - significantly reduced dimer formation (Figure 10(B)). Overall, expanding the vector genome size by incorporating additional polyA elements significantly reduced genome dimerization without compromising expression.

[0699] Example 9 - CirpT4 binding partners mass spectroscopy study

[0700] CirpT4 (SEQ ID NO: 5) was conjugated to ThermoFisher Pierce NHS magnetic beads according to manufacturer’s instructions. Unconjugated beads were prepared in the same waywithout adding CirpT4 to the reaction mixture. Beads were then incubated in vitreous humour or serum diluted in PBS at the following dilutions:

[0701] • Neat (100% matrix)

[0702] • 1 in 10 (10% matrix)

[0703] • 1 in 50 (2% matrix)

[0704] • 1 in 100 (1% matrix)

[0705] • 1 in 500 (0.2% matrix)

[0706] Beads were washed in 0.5M NaCl PBS (x2) and PBS (x2). Beads were centrifuged and collected before carrying out mass spectroscopy.

[0707] Mass spectroscopy data were analysed as follows. Peak Area data were gathered for each protein (n=836 total) across twenty samples. The data were split by matrix (vitreous and serum). Proteins with no detectable signal in a given matrix were removed. To limit the influence of extreme background signals, naive bead intensities exceeding the matrix and dilution specific 95th percentile were excluded from the noise estimation. The background noise level (limit of blank, LOB) was calculated from non-zero naive bead intensities for each matrix dilution combination. For serum, LOB was defined as the median + 1.645 mean adjusted deviation (MAD) when more than five non-zero values were available, and as the median otherwise. For the vitreous, the same criterion was applied only to neat and 1 in 10 dilutions. For higher dilutions where naive signals were sparse, the median alone was used. To ensure numerical stability, an effective LOB (LOB eff) was defined as the raw LOB when this was finite and positive or otherwise set to 1. Missing or 0 data were replaced with LOB eff. Fold-changes were calculated according to the following equation:

[0708] / Conjugated + LOBeff\

[0709] logFC = log10- - - — — - -

[0710]

[0711] \ Naive + LOBeff J

[0712] Binders were classified according to the criteria listed in Table 8.

[0713] Strict Binders At least two mid-dilutions >2-fold enrichment

[0714] At least 3 mid-dilutions present Monotonic decreasing trend across dilutions Conjugate Only Binders No naive-bead detection in any mid-dilution At least 2 conjugated detections

[0715] All mid-dilutions measurable Monotonic or decreasing trend

[0716] Weak Binders At least one mid-dilution >2-fold enrichment

[0717] Monotonic trend present

[0718] At least two mid-dilutions present

[0719] Not strict or conjugate only

[0720]

[0721] Non-binders Fails all criteria

[0722] Table 8: Binder classification criteriaCirpT4 binders were identified as set out in Table 9.

[0723] Protein Matrices Classification ATP6AP2 Vitreous Vitreous: weak

[0724] C5 Serum; vitreous Serum: strict; vitreous: weak KRT10 Vitreous Vitreous: weak

[0725]

[0726] KRT2 Vitreous Vitreous: weak

[0727] Table 9: CirpT4 binders and classification

[0728] C5 is the only strict binding partner identified for CirpT4. The remainder are identified as weak binders in the vitreous only. The non-standardised results are shown in Figures 11(A) and 11(B). Following subtraction of blank (naive beads), binders other than C5 show unreliable binding (Figures 11(C) and 11(D)). Only C5 exhibits consistent binding behaviour (e.g. not turning negative when blank subtracted). Figure 11(E) shows the binding of C5 with unreliable binders removed. C5 is classified as a weak binder in the vitreous likely due to low protein concentration in vitreous (1 concentration >2-fold-change, at least 2 mid-dilutions present, monotonic relationship).

[0729] These data show that CirpT4 has a singular binding partner (C5) in serum and vitreous.

[0730] Example 10 - Seroprevalence of Pre-Existing Anti-CirpT4 Antibodies in Human Donors A total of 100 human serum samples from US donors were acquired and tested for the presence of pre-existing antibodies against CirpT4 using the following assay. First, serial-diluted human donor samples were tested in a direct ELISA using CirpT4-coated plates. Bound anti-CirpT4 antibodies were detected with a goat anti-human IgG antibody (Figure 12). Antiserum from rabbits immunised with AAV-CirpT4 viral vector was used as a positive control. Secondly (confirmatory), all samples were reevaluated in an inhibitory ELISA in which serum samples were pre-incubated with either 100 or 200pg / mL of purified, recombinant CirpT4 protein before being applied to a CirpT4-coated plate. The supplemented CirpT4 protein concentrations were chosen to ensure that enough CirpT4 antigen is left for sequestering potential anti-CirpT4 antibodies after binding to is natural target C5 which is present in serum at concentrations of approximately 55-113 pg / mL. In the inhibitory ELISA, specific anti-drug antibody -mediated binding is demonstrated by a reduction in absorbance following competitive inhibition, whereas non-specific or matrix-related signals are not affected by the pre-incubation step. Combined results from the direct ELISA and the inhibitory assay (data not shown) indicated that none of the 100 human donor serum samples contained pre-existing antibodies to CirpT4 protein.

[0731] Example 11 - Complement-dependent Cytotoxicity assay

[0732] iLite® CD20(+) Svar Luciferase Assay Ready Cells (Svar Life Science, BM5028) derived from human Ramos B-lymphocytes (ATCC CRL-1596) and engineered for constitutive Svar luciferase expression were used to assess inhibition of complement-dependent cytotoxicity (CDC).

[0733] Cells were thawed at 37 °C and diluted according to the manufacturer’s instructions. Each well received 20 pL of 0.5 pg / mL Rituximab (final 0.18 pg / mL) followed by 20 pL of diluted cells, and plates were incubated for 30 min at 37 °C, 5% CO2. No mixing or plate shaking was performed to avoid cell damage.Three C5 inhibitors: CirpT4 (SEQ ID NO: 5) (final concentration: 50.0-0.2 pg / mL), an anti-05 antibody (final concentration: 70.0-3.1 pg / mL), and a commercially-available C5 inhibitor (final concentration: 50.0-0.2 pg / mL), were serially diluted in 1:2 serum solution and pre-incubated for 30 min at 37 °C, 5% CO2. 5 pL of each inhibitor dilution were added to the primed cells, followed by 10 pL of diluent to reach a final assay volume of 55 pL containing 4.5% human serum. Plates were incubated for 5 h at 37 °C, 5% CO2.

[0734] After incubation, plates were equilibrated to room temperature and 75 pL / well of luciferase substrate were added. Following 10 min incubation, luminescence was measured on a Varioskan LUX plate reader with 0.1 s integration time. Data were normalized to serum only control wells (100%) to calculate relative CDC inhibition.

[0735] In this assay all three C5 inhibitors effectively protected cells from complemented attack and lysis. Across three independent experiments, the mean IC50 (± SD) values were 6.5 ± 0.5 nM for CirpT4, 5.2 ± 1.1 nM for the anti-C5 antibody, and 2.3 ± 0.4 nM for the commercially-available C5 inhibitor (Figure 13).

[0736] Example 12 - In vitro membrane attack complex (MAC) deposition assay

[0737] To evaluate membrane attack complex (MAC) deposition on retinal pigmented epithelium (RPE), assays were performed using either the ARPE-19 cell line or iPSC-derived RPE (FUJIFILM CDI, 01279).

[0738] ARPE-19 cells were maintained in a 1:1 mixture of DMEM and DMEM / F12 supplemented with 10% FBS. For experiments, cells were seeded onto 24-well permeable supports (PET membrane, 0.4 pm pores, Falcon Cat. #: 353095) at 2.5 x 105 cells / well, adding 500 pL of complete medium to the basolateral chamber and 200 pL to the apical chamber. Confluent monolayers were visually confirmed the following day.

[0739] Transwells (PET, 0.4 pm; Falcon Cat. #: 353095) were coated with mouse laminin

[0740] (20 pg / mL; 50 pL / well) for 45 min at room temperature, washed with PBS, and prepared for seeding. Thawed iPSC-derived RPE progenitor cells were washed and resuspended in complete medium (MEM-a, 5% FBS, 1% N-2 supplement, 55 nM hydrocortisone, 250 pg / mL taurine, 14 pg / mL triiodo-L-thyronine, 25 pg / mL gentamicin). Cells were adjusted to

[0741] 5 x io5cells / mL and seeded at 5 x 104cells (100 pL) per apical chamber; 500 pL of medium was added basally. Differentiation proceeded for a minimum of 28 days with medium changes three times per week.

[0742] To sensitize monolayers to complement attack, cells were subjected to oxidative stress. Media were removed from the apical chamber and monolayers washed three times with 300 pL HBS+ (10 mM HEPES, 150 mM NaCl, 2 mM CaCb, 1 mM MgCb). Cells were incubated with 200 pL HBS+ containing 100 pM tert-butyl hydroperoxide (tBHP; Merck Cat. # 458139) for 1 h at 37 °C, 5% CO2.

[0743] Complement-preserved human serum (20%) was prepared in HBS+ and kept on ice. CirpT4 (SEQ ID NO: 5) was added to final concentrations of 10 nM, 20 nM, or 200 nM. After tBHP incubation, monolayers were washed twice in HBS+ and 350 pL of serum-containing HBS+ was applied to the apical compartment. Transwells were incubated for 1.5 h at 37 °C, 5% CO2.Cells were then washed three times with warm PBS (apical and basal) and fixed in 4% formaldehyde (Sigma Cat. #: 47608) in PBS for 10 min at room temperature, followed by three additional PBS washes. Inserts were stored in PBS at 4 °C until staining.

[0744] Monolayers were blocked in 10% normal goat serum (Abeam Cat. #: ab7481) in PBS for 1 h at room temperature. Primary staining was performed with mouse anti-C5b-9 (AE11; Abeam Cat. #: ab66768; 1:200) diluted in 10% goat serum and incubated for 1 h at room temperature. iPSC-RPE were additionally stained with rabbit anti-ZO-1 (Life Technologies Cat. #:

[0745] 61-7300; 1:200) to confirm differentiation status.

[0746] Secondary antibodies consisted of Alexa Fluor 594 goat anti-mouse IgG2a (Invitrogen Cat. #: A21135; 1 : 1,000) for C5b-9 and Alexa Fluor 488 goat anti-rabbit IgG (Invitrogen Cat. #: Al 1006; 1 : 1,000) for ZO-1. Samples were washed, counterstained with Hoechst (1 pg / mL, 10 min, room temperature), washed again, and stored in PBS until imaging.

[0747] Images were acquired on an EVOS M5000 microscope. Illumination settings, light intensity, and gain were optimized per channel (594 nm, 488 nm, 405 nm) and fixed per experiment. Single-channel images were captured for analysis.

[0748] MAC puncta were quantified in FIJI using a custom macro. Images from the 594 nm channel were batch-processed by conversion to 8-bit, Gaussian blur denoising, and puncta detection using Find Maxima with defined noise tolerance and background polarity settings. The macro recorded puncta counts, saved QC images with detected maxima overlays, and exported a consolidated CSV file. Data were analyzed and plotted in GraphPad Prism 7.

[0749] In ARPE 19 and iPSC derived RPE monolayers, serum dependent MAC puncta were readily detected (Figure 14(A) and 14(B)), whereas no puncta were observed in monolayers treated with heat inactivated serum. A dose dependent reduction was observed in those treated with complement inhibitors (Figure 15). Automated quantification (Figure 15(A) and 15(B)) showed a median MAC deposition of 117.5 puncta (IQR 97.75-138) in ARPE 19 cells and 78.5 puncta (IQR 73.25-114.3) in iPSC derived RPE. The addition of CirpT4 produced a clear, dose dependent reduction in MAC puncta (Figure 15(C) and (D)) across both cell types.

[0750] Example 13 - GLP-compliant Safety Study

[0751] A GLP-compliant safety (including biodistribution) study of “AAV-C5inhib_GLP” vector (CBA-CirpT4Col-WPRE-bGH-hGH-SV40) in cynomolgus macaques was carried out. A schematic diagram of AAV-C5inhib_GLP is provided in Figure 22. The sequences of the various elements of AAV-C5inhib_GLP are provided in Table 10. A detailed description of the experimental design is provided in Table 11.

[0752] Element of AAV-C5inhib GLP SEQ ID NO

[0753] Capsid 33

[0754] 5’ ITR 88

[0755] Promoter 39

[0756] CirpT4 transgene including signal peptide 20

[0757] WPRE 40

[0758] 3xPolyA (bGH+hGH+SV40) 41+42+43

[0759]

[0760] 3’ ITR 89Table 10: Sequences of AAV-C5inhib_GLP vector elements

[0761] Group Treatment Dose Dose Dose Animals (n)

[0762] No. (OU) Level Volume Concentration

[0763] (vg / eye) (pL / eye) (vg / mL) Day 91 (±1) Day 181 (±1)

[0764] Necropsy Necropsy M F M F

[0765] 1 Vehicle 0 50 0 1 2 2 1 Control

[0766] 2 AAV- 7.5 x 50 1.5 x 10122 1 1 2 C5inhib_GLP IO10

[0767] - Low Dose

[0768] 3 AAV- 2.5 x 50 5 x 10121 2 2 1 C5inhib_GLP 1011

[0769] - Mid Dose

[0770] 4 AAV- 7.5 x 50 1.5 x 10132 1 1 2 C5inhib_GLP 1011

[0771] - High Dose

[0772]

[0773] Table 11: Experimental Design of the GLP-compliant Safety Study in cynomolgus macaques

[0774] Animals received corticosteroid immunosuppression intramuscularly prior to dosing, and then weekly for four weeks, with the final dose at Week 5 (i.e., Day 35). Immunosuppression was discontinued for the remainder of the study. The safety of AAV-C5inhib_GLP was evaluated post-treatment over a 181-day observation period. For each of the treatment groups, half of the animals were euthanized on Day 91 (Week 13, n=3) and the remaining half on Day 181 (Week 25, n =3).

[0775] Safety

[0776] Overall, AAV-C5inhib_GLP demonstrated a good tolerability at doses up to 2.5 x io11vg / eye (low and mid dose); mild, self-limiting inflammation was noted by Week 25 in the 7.5 x 1011vg / eye (high dose) group. See Figures 17, 18 and 19. Up to the end of the Day 181 study period, there has been no AAV-C5inhib_GLP -related mortality or related effects on body weights, body weight gains, intraocular pressure (IOP; Figure 16), clinical pathology parameters, clinical observations, organ weights, or macroscopic findings.

[0777] There were no trends in electroretinogram (ERG) responses to suggest an AAV-C5inhib_GLP effect on any ERG parameter up to Day 181. None of the optical coherence tomography (OCT) observations reported across treatment groups were deemed related to AAV-C5inhib_GLP treatment.

[0778] By confocal scanning laser ophthalmoscope, minimal to mild multifocal perivascular hyporeflectivity within the inferior retina were observed in 5 of 12 eyes treated with 7.5 x 1011vg / eye at Day 181, indicating an increase in perivascular tissue density consistent with retinal oedema or cellular infiltrate in 5 / 6 eyes. One of 12 eyes treated with 7.5 x io11vg / eye exhibited a minimal focal hyporeflectivity within the inferior macula consistent with a retinal hemorrhage. These findings in animals treated with 7.5 x 1011vg / eye were consistent with minimal chorioretinitis.Overall, findings at the high dose were consistent with a persistent but mild immune mediated ocular response.

[0779] In conclusion, a single bilateral IVT administration of AAV-C5inhib_GLP to male and female cynomolgus macaques was well tolerated with no adverse effects up to a dose level of 7.5 x 1011vg / eye through Day 181.

[0780] Expression of CirpT4 and development of anti-CirpT4 antibodies

[0781] Expression of CirpT4 protein was measured by an in-house ELISA. All animals expressed detectable CirpT4 levels in vitreous (Figure 20(A)) and aqueous humor (Figure 20(B)) (only data from animals for which longitudinal data is available, is shown). Dose-dependent expression of CirpT4 protein was observed between treatment groups; CirpT4 protein levels were sustained in all animals up to Week 25 except for a couple of animals in the 7.5 x io11vg / eye treatment group.

[0782] Analysis of serum showed that none of the animals had pre-existing antibodies to CirpT4 protein, nor had developed any by Week 25 (data not shown).

[0783] Assessment of systemic CirpT4 activity level

[0784] To test a potential impact of circulating CirpT4, a classical pathway functional complement assay (TCC) was performed. Complement function was tested in serum samples of Cynomolgus macaques that received in each eye a single intravitreal injection of vehicle control or AAV-C5inhib_GLP at either 7.5 x 1O10(low dose), 2.5 x 1011(mid-dose), or 7.5 x 1011(high dose) vector genome / eye on Day 0. Samples were taken at Week 1 and on Terminal timepoints (Day 91 and Day 181 post-AAV-C5inhib_GLP dosing) and compared to baseline complement function of each animal. No significant change in systemic complement activity was detected (Figure 21).

[0785] Example 14 - Clinical study protocol

[0786] A phase I open label, dose escalation, multicentre study will be carried out to evaluate the safety and tolerability of AAV-C5inhib_GLP in participants with GA secondary to dry AMD. AAV-C5inhib_GLP will be administered via a single IVT injection in male and female participants who have GA secondary to dry AMD.

[0787] Approximately 13 eligible participants will be sequentially assigned to one of three cohorts. Cohort 1 (Low dose 2.5 x IO10vg per eye) may enroll a total of three participants and Cohorts 2 and 3 (Mid dose 7.5 x IO10vg and High dose 2.5 x 1011vg per eye, respectively) may enroll a total of five participants each.

[0788] Subsequent to a screening visit, and beginning with Cohort 1, AAV-C5inhib_GLP will be administered at the assigned dose level as a single dose (50 pL) intravitreal (IVT) injection in the study eye. All treated participants will undergo topical corticosteroid treatment starting 3 days prior to AAV-C5inhib_GLP administration and taper over the course of several months. Patient inclusion and exclusion criteria:

[0789] Inclusion criteria to include the following:

[0790] • >60 years of age at the time of informed consent.• Clinical diagnosis of GA secondary to AMD as per fundus autofluorescence (FAF) in one eye.

[0791] • Best-corrected visual acuity (BCVA) via the Early Treatment for Diabetic Retinopathy (ETDRS) score of <58 letters using ETDRS charts and >19 letters for the sentinel participants in each cohort and <73 letters and >34 letters for the remaining participants in each cohort. If both eyes otherwise qualify, the worse-seeing eye will be designated as the study eye. If vision is the same in both eyes, the eye with the largest total GA lesion size will be designated as the study eye as defined below. • Total GA area must be >1.25 and <20 mm2in the study eye.

[0792] o If GA is multifocal, at least one focal lesion must be >1.25 mm2, with the overall area of GA >1.25 and <20 mm2.

[0793] o Presence of hyper autofluorescence, any pattern, in the junctional zone of the GA.

[0794] o The entire GA lesion must be completely visualized on the macula centered image and must be able to be imaged in its entirety and not contiguous with any peripapillary atrophy.

[0795] Exclusion criteria to include the following:

[0796] • Complicating systemic diseases (e.g., medical conditions causing immunosuppression, active systemic infection) that would preclude the gene transfer or the IVT injection.

[0797] • Study eye:

[0798] o Absence of hyper autofluorescence.

[0799] o Prior use of any medication to treat GA, such as Syfovre® (pegcetacolpan) or Izervay® (avacincaptad pegol).

[0800] o Pre-existing eye conditions that would preclude IVT injection, interfere with the interpretation of study endpoints, or increase the risk of complications (e.g., corneal opacities, diabetic retinopathy, retinal vasculitis, active cystoid macular edema [CME]).

[0801] o History of retinal detachment.

[0802] o History or evidence of uncontrolled glaucoma or uncontrolled ocular hypertension as defined as an IOP greater than 22 mmHg on 2+ IOP lowering medications (combination medications are two or more medications by definition).

[0803] o Visual loss due to causes other than GA.

[0804] o History of vitrectomy surgery, sub-macular surgery, or other surgical interventions for AMD.

[0805] o Previous “macular” laser photocoagulation.

[0806] o Prior treatment with Visudyne® (verteporfm), external beam radiation therapy (for intraocular conditions), or transpupillary thermotherapy.

[0807] o History of intraocular inj ection within three months prior to the Screening Visit or previously received intravitreal anti-vascular endothelial growth factor injections.

[0808] o Intraocular surgery within 90 days of study administration.

[0809] o Significant media opacity impacting evaluation or imaging of the retina or vitreous. This includes cataracts considered to be a major contributor to reducing visual acuity and / or if the participant is likely to require cataract extraction within 12 months of study drug administration or during the study period.o Previous participation in interventional clinical studies, except for those only involving vitamins and minerals, irrespective of the route of administration (ie, ocular or systemic) within the last six months.

[0810] • Both eyes:

[0811] o GA secondary to a condition other than dry AMD (eg, high myopia or monogenic macular dystrophies such as Stargardt disease, cone rod dystrophy, toxic maculopathies, or myopic degeneration).

[0812] o Evidence or history of choroidal neovascularization (CNV).

[0813] o Any active ocular / intraocular infection or inflammation (eg, severe blepharitis, infectious conjunctivitis, keratitis, scleritis, endophthalmitis, idiopathic or autoimmune-associated uveitis, or herpetic lesions).

[0814] o History of steroid-induced raised intraocular pressure (IOP) of >25 mmHg following corticosteroid exposure, despite topical IOP -lowering pharmacologic therapy.

[0815] o History of artificial retinal implant or prosthesis.

[0816] o History of active bacterial, viral, fungal, or parasitic infection in the 3 months prior to the screening visit.

[0817] Patients will be assessed at Months 1 (30 days), 3, 6, 9, and 12 (and then less frequent longer-term follow-up) following administration of the study drug in order to monitor progress of the study eyes.

[0818] Primary and secondary endpoints to include the following:

[0819] • The number and proportion of participants experiencing severe ocular and / or nonocular treatment-emergent adverse events (TEAEs), including serious adverse events (SAEs) through Month 12.

[0820] • The number and proportion of participants experiencing ocular and / or non-ocular TEAEs, including treatment-emergent SAEs.

[0821] • Change in the area of GA lesion size, as measured by FAF.

[0822] • Change in EZ attenuation and area as measured by optical coherence tomography (OCT).

[0823] • Change from baseline in BCVA via the ETDRS chart.

[0824] • Change from baseline in low-luminance visual acuity (LEVA) via the ETDRS chart.

[0825] • Change from baseline on the Functional Reading Independence Index (FRII).

[0826] • Change from baseline in reading performance as assessed by Minnesota Low Vision Reading Test (MNRead) chart.

[0827] • Change in antibody titre to AAV6 derived capsid, CirpT4, and T-cell response to AAV-6 derived capsid and CirpT4.

Claims

1. CLAIMS1. A method of treating an ocular disorder comprising a step of intravitreal administration of a vector comprising a recombinant genome comprising a polynucleotide comprising a transgene encoding a C5 inhibitor.

2. The method of claim 1, wherein the C5 inhibitor:(a) can bind to C5, and / or prevent cleavage of C5 to C5b by C5 convertase;(b) can reduce TCC formation by at least 50%, or at least 70% of the level of reduction shown by a C5 inhibitor consisting of the amino acid sequence of SEQ ID NO: 5 in an equivalent complement activity assay;(c) is a C5 inhibitor that can bind to the peripheral macro globulin domain 4 (C5 MG4) and / or peripheral macro globulin domain 5 (C5 MG5) of C5; and / or(d) is a polypeptide from the CirpT family, optionally wherein the C5 inhibitor is a polypeptide selected from the group consisting of a CirpTl polypeptide, a CirpT2 polypeptide, a CirpT3 polypeptide, and a CirpT4 polypeptide.

3. The method of claim 1 or claim 2, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence:(a) of any one of SEQ ID NOs: 6, 7, 84 or 85;(b) which is:(i) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 50, at least 60, at least 70, or at least 80 amino acids of any one of SEQ ID NOs: 1-5;(ii) at least 95% identical to a fragment of at least 80 amino acids of any one of SEQ ID NOs: 1-5;(iii) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 1-5; or(iv) identical to any one of SEQ ID NOs: 1-5; and / or(c) which is:(i) identical to SEQ ID NO: 5, but for one or more conservative amino acid substitutions at one or more positions corresponding to positions 1, 2, 3, 5, 8, 12, 18, 22, 23, 28, 30, 37, 42, 43, 51, 54, 60, 67, 78, 83 and 88 of SEQ ID NO: 5; and / or (ii) identical to SEQ ID NO: 5, but for one or more conservative amino acid substitutions corresponding to one or more of DIE, I2V, Q3R, R5H, S8T, R12K, N18D, 122 V, F23Y, L28I, D30N, H37N, I42L, A43S, E51K, A54S, F60I, R67H, Q78N, Q83H, and T88A.

4. The vector of any one of the preceding claims, wherein the C5 inhibitor comprises a polypeptide comprising an amino acid sequence which is at least 95% identical to SEQ ID NO: 5.

5. The method of any one of the preceding claims, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 95% identical to any one of SEQ ID NOs: 21 and 44 to 46.

826. The method of any one of the preceding claims, wherein the polynucleotide and / or the transgene(a) is codon optimised; and / or(b) comprises a reduced number of CpGs compared to a corresponding portion of the corresponding wild type nucleotide sequence, optionally wherein the polynucleotide and / or the transgene is CpG-free.

7. The method of claim 6, wherein the polynucleotide and / or the transgene comprises a nucleotide sequence which is at least 98% identical to any one of SEQ ID NOs: 54 to 56.

8. The method of any one of the preceding claims, wherein the polynucleotide comprises a nucleotide sequence which is:(a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 175, at least 200, at least 225, or at least 250 nucleotides of any one of SEQ ID NOs: 22, 23, 25 or 26;(b) at least 95% identical to a fragment of at least 250 nucleotides of any one of SEQ ID NOs: 22, 23, 25 or 26;(c) at least 90%, at least 95%, or at least 98% identical to any one of SEQ ID NOs: 22, 23, 25 or 26; and / or(d) identical to any one of SEQ ID NOs: 22, 23, 25 or 26.

9. The method of any one of the preceding claims, wherein the C5 inhibitor comprises a signal peptide, optionally a human protein-derived signal peptide, further optionally an alpha-2-HS-glycoprotein / fetuin-A (AHSG) signal peptide.

10. The method of claim 9, wherein the signal peptide comprises an amino acid sequence:(a) at least 90%, or at least 95% identical to SEQ ID NO: 13;(b) identical to SEQ ID NO: 13 but for one, two, or three amino acid substitutions; and / or(c) identical to SEQ ID NO: 13.

11. The method of claim 9 or claim 10, wherein the polynucleotide comprises a nucleotide sequence which is:(a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 200, at least 225, at least 250, at least 275, or at least 300 nucleotides of SEQ ID NO: 20; (b) at least 95% identical to a fragment of at least 300 nucleotides of SEQ ID NO: 20;(c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 20; and / or (d) identical to SEQ ID NO: 20.

12. The method of any one of the preceding claims, wherein the vector is an AAV particle comprising a capsid, optionally wherein the capsid is:(a) an AAV2 capsid;(b) an AAV2-derived capsid;(c) an AAV6 capsid; or(d) an AAV6-derived capsid.

13. The method of claim 12, wherein the capsid comprises a polypeptide comprising an amino acid sequence at least 90%, at least 95%, at least 98%, at least 99%, or at least 99.5% identical to a fragment of at least 600, at least 650, at least 675, or at least 700 amino acids of any one of SEQ ID NO: 32-34, preferably SEQ ID NO: 33.

14. The method of any one of the preceding claims, wherein the polynucleotide comprises a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), optionally wherein the WPRE comprises a polynucleotide comprising a nucleotide sequence which is:(a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, or at least 550 nucleotides of SEQ ID NO: 40;(b) at least 95% identical to a fragment of at least 550 nucleotides of SEQ ID NO: 40;(c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 40; or (d) identical to SEQ ID NO: 40.

15. The method of any one of the preceding claims, wherein the polynucleotide comprises a CBA promoter, optionally wherein the CBA promoter comprises a polynucleotide comprising a nucleotide sequence which is:(a) at least 90%, at least 95%, or at least 98% identical to a fragment of at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, or at least 800 nucleotides of SEQ ID NO: 39;(b) at least 95% identical to a fragment of at least 800 nucleotides of SEQ ID NO: 39;(c) at least 90%, at least 95%, or at least 98% identical to SEQ ID NO: 39; or (d) identical to SEQ ID NO: 39.

16. The method of any one of the preceding claims, wherein the polynucleotide comprises a 5’ ITR and a 3’ ITR, optionally wherein:(a) the 5’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 88 or 93; and / or(b) the 3’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 89 or 94.

17. A vector comprising a recombinant genome comprising, in 5’ to 3’ order, a 5’ ITR, a promoter, a transgene encoding a C5 inhibitor, a WPRE sequence, three polyA sequences, and a 3’ ITR.

18. The vector of claim 17, wherein:(a) the promoter comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 39;(b) the transgene comprises a nucleotide sequence which is at least 98% identical or identical to SEQ ID NO: 20;(c) the WPRE sequence comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 40;(d) the three polyA sequences comprise a nucleotide sequence comprising (a) a sequence which is at least 95% identical, at least 98% identical or identical to SEQ ID NO: 41; (b) a sequence which is at least 95% identical, at least 98% identical or identical to SEQ ID NO:42; and (c) a sequence which is at least 95% identical, at least 98% identical or identical to SEQ ID NO: 43;(e) the 5’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 88; and / or(f) the 3’ ITR comprises a nucleotide sequence which is at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 89.

19. The method of any one of claims 1 to 16, wherein the ocular disorder is a complement-mediated disorder, a disorder associated with C5 and / or a condition that would benefit from reducing complement-mediated inflammation.

20. The method of any one of claims 1 to 16 or 19, wherein the ocular disorder is age-related macular degeneration (AMD) or geographic atrophy (GA).85

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