Compositions and methods for SPG11 gene therapy

Abstract

Lentiviral compositions comprising SPG11 are provided herein. The lentiviral compositions are useful in methods of treating lysosome storage related disorders including but not limited to SPG11 related disorders. Methods of treating a patient have a condition associated with SPG11 mutation are provided.

Description

[0001] COMPOSITIONS AND METHODS FOR SPG11 GENE THERAPY

[0002] Cross-Reference

[0003] This application claims the benefit of U.S. Provisional Application No. 63 / 661971, filed June 20, 2024, each of which is incorporated herein by reference in its entirety.

[0004] Sequence Listing Incorporation By Reference

[0005] The application herein incorporates by reference in its entirety the sequence listing material submitted concurrently with the specification as an XML file, with a filename of “23963001W01”, a production date of June 11, 2025, and a size of 49,152 bytes. The ST26 Sequence Listing is part of the specification and is incorporated in its entirety by reference herein.

[0006] BACKGROUND

[0007] SPG11 is the gene encoding for spatacsin. Mutations in the SPG11 gene can result in accumulation of lipids within lysosomes, and result in a broad array of symptoms associated with spastic paraplegia, among other disease conditions. In particular, loss of functional spatacsin is believed to lead to lysosomal dysfunction by perturbing the clearance of lipids such as gangliosides from lysosomes. This lysosomal dysfunction can trigger neuronal cell death.

[0008] There are currently no genetic therapies for SPG11 -related conditions. Ameliorative care for spastic paraplegia, for example, involves physiotherapy and antispastic drugs. Genetic therapies to stop progression or reverse these conditions are needed.

[0009] The coding sequence for the SPG11 gene is approximately 8.5 kilobases. Virus-like particles (VLPs) have a packaging capacity of approximately 4 to 6 kilobases. The dual adeno-associated virus (AAV) intein splicing system requires co-transduction and proper splicing in the same cell. Current AAV vectors used in gene therapy are more than a decade old. Lentiviral vectors remain near the injection site. 1 DESCRIPTION OF THE FIGURES

[0010] 2 FIG. 1 illustrates lysosomal dysfunction caused by SPG11 mutation. From Boutry,

[0011] 3 M. et al. Inhibition of Lysosome Membrane Recycling Causes Accumulation of

[0012] 4 Gangliosides That Contribute to Neurodegeneration. Cell Reports 23 (13): 3813-26 (2018).

[0013] 5 FIG. 2 shows a proof of concept of an SPG11 gene therapy. Patient-derived induced

[0014] 6 pluripotent stem cells (iPSCs) were differentiated into neurons and treated with a lentiviral

[0015] 7 vector carrying the SPG11 gene. Cells were visualized using LysoTracker Red before

[0016] 8 (Diseased Neuron: Pre-treatment) and after treatment (Corrected Neuron: Post treat) with

[0017] 9 the SPG11 -carrying lentiviral vector to visualize lysosomes. Diseased neuronal cells with

[0018] 10 high lysozyme levels are visible prior to treatment. After treatment with lentivirus carrying

[0019] 11 SPG11, the staining is substantially reduced.

[0020] 12 FIG. 3 provides micrographs of frozen induced pluripotent stem cells (iPSC)

[0021] 13 developed from PBMC isolated from whole blood collected from a diseased patient.

[0022] 14 Figure 4 provides an overview of the LysoTracker Red Assay for neural stem cells.

[0023] 15 FIG. 4A provides a panel of 8 micrograph images for red / blue area analysis. FIG. 4B

[0024] 16 provides a panel of 8 micrograph images with filter allowing the areas retaining the

[0025] 17 LysoTracker Red signal to be measured. FIG. 4C provides a panel of 8 micrograph images

[0026] 18 with filter allowing the filters with areas retaining the DAPI signal to be measured. The ratio

[0027] 19 of red area / blue area = relative transduction.

[0028] 20 Figure 5 summarizes results obtained from neuronal stem cells (NSC) derived from

[0029] 21 cells obtained from a patient with lysosome storage disorder either untreated or treated with

[0030] 22 lentivirus vectors comprising the SPG11 gene at varying multiplicities of infection (MOI).

[0031] 23 FIGS. 5A-5E contain micrographs of the cells. FIG. 5A shows untreated NSC cells.

[0032] 24 Untreated cells demonstrate a high ratio of nearly 18% red to blue ratio demonstrating

[0033] 25 profound lysosomal storage disease. FIG. 5B shows NSC cells treated with lentivirus vector

[0034] 26 comprising the SPG11 gene at an MOUL The rediblue ratio is 0.0234. FIG. 5C shows

[0035] 'Ll NSC cells treated with lentivirus vector comprising the SPG11 gene at an MOI=2. The

[0036] 28 red:blue ratio is 0.0247. FIG. 5D shows NSC cells treated with lentivirus vector comprising

[0037] 29 the SPG11 gene at an MOI=5. The red:blue ratio is 0.0287. FIG. 5E shows NSC cells 1 treated with lentivirus vector comprising the SPG11 gene at an MOI=10. The red:blue ratio

[0038] 2 is 0.0653. FIG. 5F provides a chart summarizing the results obtained from lentiviral SPG11

[0039] 3 vector treatment of NSC from a patient at the indicated MOI. Untreated healthy donor NSCs

[0040] 4 (Unrelated donor) and untreated CRISPER Corrected NSCs from a second patient (MD

[0041] 5 CRISPR Corrected) were evaluated in the Lysotracker Assay-Red DAPI-blue assay also,

[0042] 6 and summary results are included in the chart.

[0043] 7 DESCRIPTION OF EMBODIMENTS

[0044] 8 In the various aspects and embodiments, this disclosure provides a therapeutic

[0045] 9 strategy for patients suffering from a condition or disease relating to SPG11 mutation, and

[0046] 10 which can include lysosomal dysfunction. Diseases linked to SPG11 mutations or the lack

[0047] 11 of functional SPG11 genes are characterized by a range of severe symptoms, notably

[0048] 12 neurodegeneration. In embodiments, the disclosure provides lentiviral vectors to introduce

[0049] 13 a functional SPG11 gene into the cells of individuals impacted by such conditions, to correct

[0050] 14 abnormal sphingolipid levels and restore lysosomal health. In embodiments, the disclosure

[0051] 15 provides symptomatic relief, and in embodiments slows, halts, or reverses disease

[0052] 16 progression.

[0053] 17 The spastic paraplegia 11 (SPG11) gene encodes a 40 exon, 8-kb mRNA (encoding

[0054] 18 2443 amino acids, (SEQ ID NO: 1)) that is broadly expressed in cells and tissues. Within the

[0055] 19 brain, SPG11 is expressed, for example, in the adult cerebellum, cerebral cortex,

[0056] 20 hippocampus, and pineal gland. Diverse clinical features have been reported with SPG11

[0057] 21 mutations, and these include diseases recognized as hereditary spastic paraplegia (HSP),

[0058] 22 Kjellin syndromejuvenile amyotrophic lateral sclerosis, and Parkinsonism.

[0059] 23 HSP is characterized by progressive spasticity and weakness of the lower limbs, and

[0060] 24 can be associated with mild intellectual disability in childhood and / or progressive cognitive

[0061] 25 decline, as well as peripheral neuropathy, pseudobulbar involvement, and increased reflexes

[0062] 26 in the upper limbs. HSP can also be associated with cerebellar signs (ataxia, nystagmus,

[0063] 'Ll saccadic pursuit), retinal degeneration, pes cavus, scoliosis, amyotrophy, and parkinsonism.

[0064] 28 Onset of HSP usually occurs during infancy or adolescence. Most affected individuals are

[0065] 29 wheelchair bound within one or two decades after disease onset. Approximately ten years

[0066] 30 after onset, most affected individuals have the complete clinical picture of HSP, including progressive lower-limb spasticity, atrophy of the corpus callosum with intellectual disability, and / or progressive cognitive decline. Thinning of the corpus callosum often correlates with disease severity.

[0067] The diagnosis of SPG11-related disorder can be confirmed by molecular genetic testing. SPG11 mutations are inherited in an autosomal recessive manner. In embodiments, the patient has at least one pathogenic SPG11 mutation in one or both alleles. In embodiments, the mutations are compound heterozygous mutations. Without limitation, pathogenic mutations may be in introns or exons (e.g., exons 1 to 40). In exemplary embodiments, at least one pathogenic mutation is in exon 1, 3, 4, 6, 7, 8, 10, 11, 12, 15, 16, 25, 30, 31, 32, 34, 36, or 37. In various embodiments, the pathogenic mutation is a loss-of- function mutation, and may cause one or more of a frameshift, truncation (e.g., premature stop or nonsense mutation), an amino acid change, and deletion or insertion of one or more amino acids.

[0068] As used herein, the term “SPG11 gene” and similar terms used in connection with virus vector delivery and expression refers to a polynucleotide (DNA) that comprises an SPG11 coding sequence that does not include any pathogenic mutations, and which may code for the amino acid sequence ofSEQ ID NO: 1. The term “SPG11 gene” may be a cDNA sequence (as represented by SEQ ID NO: 2), and which may be codon optimized (an example of which is provided as SEQ ID NO: 3), and / or may comprise one or more introns, which in embodiments may impart some gene expression regulation. For example, the full gene sequence with introns and splice variants are provided by gene Reference: ENSG00000104133. The term “SPG11 gene” includes naturally occurring, non-pathogenic nucleotide polymorphisms in the human population that are not associated with disease or loss of SPG11 function or expression. In various embodiments, the SPG11 gene can be a functional equivalent of the SPG11 cDNA provided herein as SEQ ID NO: 2 or 3, that is, the SPG11 gene may encode one or more amino acid modifications (such as from one to ten, e.g., one, two, three, four, or five amino acid modifications) independently selected from amino acid insertions, deletions, or substitutions, and which do not significantly impact SPG11 activity. Generally, a functional derivative will encode an amino acid sequence having at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 1. A SPG1 1 gene may further comprise regulatory elements, including 5'- and 3'- untranslated regions, although these regulatory elements are not restricted to the naturally occurring SPG11 gene sequences, but instead can be selected to achieve the desired level of mRNA expression or turnover. In some embodiments, the SPG11 gene does not include substantial untranslated regions, that is, may consist essentially of or consist of the SPG11 coding sequence to reduce the size of the cargo for packaging into the viral vector. For example, in embodiments, the SPG11 gene contains less than 500 base pairs, or less than 400 base pairs, or less than 250 base pairs, or less than 100 base pairs of nucleotide sequence that is transcribed but untranslated.

[0069] As used herein, the terms “sequence identity”, “percent identity” and “percent sequence identity” refer to the degree of identity between two or more polynucleotides when aligned using a nucleotide sequence alignment program; or between two or more polypeptide sequences when aligned using an amino acid sequence alignment program. Similarly, the terms “identical” and percent “identity” when used herein in the context of two or more nucleotide or amino acid sequences refers to two sequences that are the same or have a specified percentage of amino acid residues or nucleotides when compared and aligned for maximum correspondence, for example as measured using a sequence comparison algorithm, e.g., the Smith -Waterman algorithm, etc. or by visual inspection. The percent identity between amino acid sequences may be determined using by, for example, the Needleman and Wunsch (1970, J. Mol. Biol. 48:444-453) algorithm which has been incorporated into the GAP program in the GCG software package, using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6 or 4 and a length weight of 1 , 2, 3, 4, 5, or 6. As another example, the percent identity between two nucleotide sequences may be determined using the GAP program in the GCG software package, using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and length weight of 1, 2, 3, 4, 5 or 6. A particularly preferred set of parameters (and the one that should be used unless otherwise specified) are a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid or nucleotide sequences may also be determined using the algorithm of E. Meyers and W. Miller (1989, Cabios, 4: 11-17) which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12, and 1 a gap penalty of 4. Additional search and alignment tools known in the art include, but are

[0070] 2 not limited to, the NBLAST and XBLAST programs (version 2.0) of Altschul et al (1990)

[0071] 3 J. Mol. Biol. 215:403-410 and the Gapped BLAST program.

[0072] 4 In accordance with embodiments of the disclosure, the SPG11 gene is under control

[0073] 5 of a promoter, which in various embodiments is a heterologous promoter. As used herein, a

[0074] 6 “heterologous promoter” is a promoter that is placed in a non-native location, that is, in

[0075] 7 position to control expression of a coding sequence that it does not control in nature. As

[0076] 8 used herein, the terms “under control of’ and “operably linked” refers to a juxtaposition of

[0077] 9 genetic elements, e.g., promoter, enhancer, termination signal sequence, polyadenylation

[0078] 10 sequence, Kozak sequence, etc., wherein the elements are in a relationship permitting them

[0079] 11 to operate in the expected manner. For instance, a promoter is operably linked to a coding

[0080] 12 region if the promoter helps initiate transcription of the coding sequence. There may be

[0081] 13 intervening residues between the promoter and the coding sequence or between any two

[0082] 14 elements so long as the functional relationship is maintained.

[0083] 15 In embodiments, the SPG11 gene is a codon-optimized cDNA coding for SPG11

[0084] 16 protein (e.g., SEQ ID NO: 1). Codon optimization involves modifying the DNA sequence

[0085] 17 to use codons that are more frequently used in the target organism (e.g., human), enhancing

[0086] 18 the efficiency of protein synthesis. This optimization process can significantly improve the

[0087] 19 expression level of the SPG11 protein in human cells, potentially increasing the therapeutic

[0088] 20 benefits. In embodiments, the codon-optimization improves the efficiency of transcription,

[0089] 21 and / or mRNA folding in a manner that improves transcription, mRNA stability, and / or

[0090] 22 translation.

[0091] 23 In some embodiments, the SPG11 gene encodes the amino acid sequence of SEQ ID

[0092] 24 NO: 1. In some embodiments, the SPG11 gene is a cDNA sequence, which in some

[0093] 25 embodiments comprises the nucleotide sequence of SEQ ID NO: 2 or 3.

[0094] 26 The term “promoter” refers to a DNA sequence capable of controlling the expression

[0095] 'Ll of an mRNA, such as transcription of the SPG11 gene. Promoter sequences contain at least

[0096] 28 proximal elements for controlling gene expression, and may optionally further comprise

[0097] 29 more distal upstream elements, the latter elements often referred to as enhancers.

[0098] 30 Accordingly, an “enhancer” is a DNA sequence that can stimulate promoter activity, or is an innate element of the promoter or a heterologous element inserted to enhance the level or tissue specificity of a promoter. Promoters may be derived in their entirety from a native gene, or may be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments. It is further recognized that the exact boundaries of regulatory sequences may not be completely defined, and thus DNA fragments of some variation may have identical promoter activity.

[0099] In embodiments, the SPG11 gene is under the control of a ubiquitous promoter (e.g., a promoter that broadly drives expression in many human tissues and cell types), such as the human cytomegalovirus (CMV) immediate early promoter, EFl -alpha (EFla) promoter, or promoter of UBC. In some embodiments, the promoter is a strong ubiquitous promoter, such as EFla promoter. Other ubiquitous promoters include 13-actin promoter, GAPDH promoter, and HSP70 promoter, among others known to the skilled person. The ubiquitous promoter can be selected to provide a level of expression similar to native SPG11 in core tissues or cells (e.g., neurons). In embodiments, the promoter is the SPG11 endogenous promoter or core promoter elements thereof. In embodiments, the promoter is selected for its ability to initiate transcription in a wide array of cell types, ensuring the broad expression of the SPG11 functional gene across different tissues affected by the disorder. In embodiments, the promoter provides expression in the brain (e.g., in neurons). In embodiments, the promoter is EFla promoter. The phrase “not from the natural SPG11 gene” is intended to encompass any sequence not typically associated with an unmodified H. sapiens SPG11 gene.

[0100] In embodiments, the promoter directs expression of the SPG11 gene at least in neurons, and in embodiments is a neuron-specific promoter such as synapsin I promoter, CamKII promoter, MeCP2 promoter, NSE promoter, and Hb9 promoter, or a derivative thereof. In embodiments, the promoter provides expression in other cells of the brain or CNS, such as glial cells. In embodiments, the promoter allows expression in oligodendrocytes and / or astrocytes. Exemplary promoters include, but are not limited to, MBP promoter, PLP1 promoter, CNP promoter, GFAP promoter, SI 00b promoter, or a derivative thereof. By “neuron-specific promoter” is intended a promoter that preferentially functions in neuron type cells versus other cell types; the preferential functioning may be an increased activity in neuron-specific cell as compared to in a non-neuron type cell. In embodiments, the promoter comprises core elements of the SPG1 1 endogenous promoter, and optionally comprises one or more enhancer sequences, which can comprise distal portions of the SPG11 endogenous gene or other promoter (e.g., a ubiquitous promoter or neuron- specific promoter). In embodiments, the SPG11 expression cassette comprises at least 200 bps, or at least 300 bps, or at least 500 bps, or at least 800 bps, or at least 1000 bps, or at least 1500 bps upstream from the SPG11 transcription start site. The 1000 bases upstream of the SPG11 transcription start site is provided herein as SEQ ID NO: 4.

[0101] In various embodiments, the SPG11 gene comprises untranslated sequences (e g., a 3'-UTR) that enhance mRNA stability. In embodiments, the SPG11 gene comprises a polyadenylation signal. A polyadenylation signal is included to ensure proper RNA processing and stability. The presence of a polyadenylation signal is crucial for the termination of transcription, the addition of a poly(A) tail to the mRNA, and subsequent stability and export of the mRNA from the nucleus. In embodiments, the polyadenylation signal is derived from the bovine growth hormone (BGH) gene or simian virus 40 (SV40), providing reliable mRNA processing and stability in a wide range of mammalian cells.

[0102] In some embodiments, the SPG11 gene is delivered with one or more detectable labels, including but not limited to a fluorescent protein, such as a GFP or RFP, allowing for visualization of expression of the SPG11 -containing expression construct.

[0103] In embodiments, the viral vector is a lentivirus vector (LV) comprising an SPG11 gene operatively linked to a promoter. The lentivirus vector comprising the SPG11 gene may be used for delivering an SPG11 gene into patient cells. In embodiments, the viral vector is a lentivirus vector (LV) delivering an SPG11 gene into patient cells. The lentivirus vector is a replication-defective retroviral vector that integrates into the host cell genome. The term “lentivirus vector” is intended to encompass wild-type and modified lentivirus vectors. Lentivirus contains an RNA genome that is reverse-transcribed into a double-strand DNA molecule, which is then stably integrated into the host cell genome mediated by the viral pre-integration complex (PIC) and the integrase protein (IN). Once integrated in the host genome, the provirus is composed of two long terminal repeats (LTRs) flanking the structural and accessory genes and sequences necessary for reverse transcription and packaging. The LTRs are tripartite elements containing essential regulatory sequences: a sequence at the 5' end (U5) contains the polyadenylation signal, followed by a repeated sequence (R) used as primer during reverse transcription and a large sequence at the 3' end (U3) containing the viral promoter and enhancer elements binding host cell transcription factors. Simple retroviruses contain only four fundamental genes, gag, pro, pol and env, necessary for the viral particle production, assembly and post-entry processing eventually leading to proviral integration. However, lentiviruses contain other accessory genes for regulating viral transcription, RNA processing, nuclear entry, and viral-host interactions. The design of gene transfer vectors from lentiviruses involves the replacement of viral genes with a therapeutic gene expression cassette (e.g., comprising a SPG11 functional gene), with retention of only the sequences necessary for vector packaging, reverse transcription, and integration. Modified lentivirus vectors may include, but are not limited to, modified lentivirus capsids such as those with improved ability to cross the blood-brain barrier, capsids with improved tropism for target cells of interest, lentiviral pseudotypes and integrase deficient recombinant lentiviruses (IDLV). Any suitable lentiviral pseudotype known in the art may be used in the compositions and methods of the invention.

[0104] Lentiviral pseudotypes include, but are not limited to, pseudotypes comprising VSV- G, BaEV, NiV, SeV, ASLV, EnvA / EnvB, LCMV, MuLV, RV-G, fusion envelope glycoproteins comprising RV-G and VSV-G segments, Hi-Ret, NeuRet, modified Sinbis envelope, a chimeric envelope comprising a growth factor, single-chain antibody variable fragments (scFvs), and combined fragments from different viral envelopes. Growth factors may include, but are not limited to IGF-I, EGF, EPO and SDF-la. Combined fragments from different envelopes may include, but are not limited to, GALV-Env and GALV- C4070A.

[0105] The main viral determinants of the integration process are the PIC, with its major functional component, the IN protein, and the IN binding sites in the LTRs. The IN protein is specific to each retrovirus type and is responsible for most of its integration preferences. Since IN is an indispensable component of a retroviral vector packaging process, it will accurately reproduce in the vector the integration characteristics of the parental virus. Lentiviral PIC enters the nucleus by an active import mechanism allowing a lentivirus to transduce both dividing and non-dividing cells. In some embodiments, the lentiviral vector is derived from HIV-1 , and which integrates in actively transcribed genes. The integration pattern is therefore determined by the specific transcriptional program of the host cell.

[0106] In embodiments, LVs are produced as replication-defective viral particles in which the RNA genome contains a so-called self-inactivating (SIN) 3 ' LTR, which upon reverse transcription into a linear, double-stranded DNA genome gives rise to an U3-deleted, enhancer- and promoter-less 5' LTR. The AU3 LTRs maintain a minimal, 18-bp long HIV- 1 sequence necessary for integration, plus the R and U5 region containing the vector polyadenylation sequence. A SIN LV provirus is transcriptionally inactive, allowing the vector to carry a fully independent transgene expression cassette. In addition, a LV genome maintains the cis-acting viral sequences necessary for encapsidation, reverse transcription and integration in the host cell genome, and lacks all other viral regulatory elements and genes, replaced by the transgene cargo (e.g., SPG11 expression cassette). These necessary sequences are the packaging signal (T), the primer binding site (PBS) and polypurine tract (PPT) required for reverse transcription, the major HIV-1 intron with donor and acceptor splice sites, and the Rev-responsive element (RRE).

[0107] Packaging a vector genome is achieved by complementation in trans of all necessary viral functions in the context of a packaging cell, where the genes encoding for these functions are transfected as independent plasmids. In embodiments, the packaging cells contain two plasmids, the first expressing the gag, pol, vif, vpr, vpu, nef, tat and rev genes and a second expressing an env gene. In embodiments, the HIV-1 env gene is replaced, for example, with a coding sequence for the glycoprotein from bovine vesicular stomatitis virus (VSV-G), which binds the ubiquitous low-density lipoprotein (LDL) receptor. VSV-G pseudotyping expands the LV tropism and enables the production of high-titer LV preparations. In embodiments, the packaging system is devoid of all HIV accessory genes and designed to package SIN vector transcribed from a heterologous promoter, such as cytomegalovirus (CMV) promoter, in a Tat-independent fashion. This system improves the biosafety of viral production by splitting the structural gag / pol and env genes into two independent plasmids to reduce the chances of generating replication-competent lentiviruses by plasmid recombination during packaging. In embodiments, the LV carries and expresses an SPG11 gene in the form of an independent expression cassette, containing an intron-less protein-coding region (cDNA) transcribed by a promoter and accessory regulatory regions and polyadenylated at the viral signal located in the 3' LTR. The desired regulation of transgene expression can be obtained at the level of transcription by the use of tissue-, cell type- or differentiation stage-specific promoters / enhancers, and / or post-transcriptionally, by adding 5' or 3' untranslated regions (UTRs) enhancing ribosome binding or mRNA stability or target sequences for specific micro RNAs (miRNAs) to regulate protein expression by physiological RNA interference mechanisms.

[0108] The size, and most importantly the sequence of the elements introduced in an LV can have a negative impact on vector titer and infectivity. As a general rule, the longer is an expression cassette, the higher is the likelihood that an LV will have low titer and / or infectivity. However, the complexity and the nature of the incorporated sequences can have an impact on vector performance. During packaging, cryptic splicing and polyadenylation signals or repeated sequences inadvertently introduced in the sense strand of the LV may slow down transcription of the vector plasmid or produce defective or prematurely terminated vector genomes. Defective genomes will reduce the proportion of vectorcontaining virions in an LV preparation or cause defective or incomplete reverse transcription upon infection of the target cell. Reverse transcription is a crucial step in LV transduction, and any sequence that slows down or halts progression of the HIV-1 reverse transcriptase will ultimately reduce the amount of double-stranded linear genomes that can be integrated in the target cell. Defects in genome packaging or integration will read out as low titer in a conventional infectious titer assay. In embodiments, the constructs provided herein produce high titer of LV particles for therapy.

[0109] In embodiments, the lentivirus vector comprises a polyadenylation signal following the SPG1 1 cDNA sequence. A polyadenylation signal is included to ensure proper RNA processing and stability. The presence of a polyadenylation signal is crucial for the termination of transcription, the addition of a poly(A) tail to the mRNA, and subsequent stability and export of the mRNA from the nucleus. In embodiments, the polyadenylation signal is derived from the bovine growth hormone (BGH) gene or simian virus 40 (SV40), providing reliable mRNA processing and stability in a wide range of mammalian cells.

[0110] An exemplary cassette for preparing lentivirus vector harboring the SPG11 gene under control of a promoter is provided herein as SEQ ID NO: 5 (EFla promoter) and SEQ ID NO: 6 (UBC promoter). Constructs can be driven by a suitable promoter (e.g., CMV promoter) for expression and packaging of the vector RNA using a producer strain, which are commercially available. In embodiments, the SPG11 gene is under the control of an EFl a promoter, which may comprise at least 200 bps, or at least 400 bps, or at least 500 bps, or at least 750 bps of the EFla promoter included in SEQ ID NO: 5. In embodiments, the SPG11 gene is under the control of the EFla promoter included in SEQ ID NO: 5. In embodiments, the SPG11 gene is under the control of a UBC promoter, which may comprise at least 200 bps, or at least 400 bps, or at least 500 bps, or at least 750 bps of the UBC promoter included in SEQ ID NO: 6. In embodiments, the SPG11 gene is under the control of the UBC promoter included in SEQ ID NO: 6.

[0111] Upon administration, the lentivirus efficiently transduces target cells, integrating the therapeutic SPG11 gene into the host genome. Without being limited by mechanism, this leads to the stable, long-term expression of SPG11, correcting the deficiency responsible for the accumulation of sphingolipids and the subsequent lysosomal storage disorder. The use of a ubiquitous promoter as described ensures that the therapeutic gene is expressed in a wide range of cell types, including those most affected by the disorder, such as neurons and other cell types within the central nervous system.

[0112] In some embodiments, the individual to be treated is a pediatric or neonatal patient (e.g., a patient with HSP). In some embodiments, early treatment avoids manifestation of some clinical parameters of disease, such as neurodegeneration, spasticity and weakness in lower or upper limbs, intellectual disability, peripheral neuropathy. In some embodiments, early treatment slows or halts disease progression. In embodiments, treatment prevents substantial or significant thinning of the corpus callosum. In some embodiments, the individual is an adult patient (e.g., with HSP), and treatment can ameliorate one or more of the clinical parameters. In various embodiments, the individual or patient for treatment exhibits one or more of the clinical symptoms selected of HSP. 1 In embodiments, the individual to be treated exhibits clinical symptoms of Kjellin

[0113] 2 syndrome, juvenile amyotrophic lateral sclerosis, and parkinsonism. In embodiments, the

[0114] 3 individual for treatment carries pathogenic mutations in both SPG11 genes, that is, mutations

[0115] 4 associated with disease. Pathogenic mutations can be as already described, and may include

[0116] 5 a loss-of-function mutation, and may cause one or more of a frameshift, truncation (e g.,

[0117] 6 premature stop or nonsense mutation), an amino acid change, and deletion or insertion of

[0118] 7 one or more amino acids.

[0119] 8 In embodiments, the individual for treatment does not show any signs of clinical

[0120] 9 symptoms, but carries pathogenic mutations in both SPG11 genes, that is, mutations

[0121] 10 associated with disease.

[0122] 11 Routes of administration of the viral vector may be systemic delivery; or may be

[0123] 12 systemic in connection with various manipulations (such as microbubble-enhanced

[0124] 13 diagnostic ultrasound (MEUS), transcranial magnetic stimulation (TMS)) to transiently open

[0125] 14 the blood brain barrier. In other embodiments, delivery is by nasal administration. In some

[0126] 15 embodiments, the viral vector is administered by systemic deliver. In some embodiments,

[0127] 16 the viral vector is administered to the brain or CNS.

[0128] 17 In still other embodiments, delivery to the brain is by delivery to the cerebrospinal

[0129] 18 fluid. Another option is delivery to the cistema magna route of injection, which is an

[0130] 19 alternative method for delivery into cerebrospinal fluid (CSF) which results in wide-spread

[0131] 20 gene delivery throughout the CNS. In some embodiments, the administration is by direct

[0132] 21 injection into the parenchyma, or by injection into the cerebrospinal fluid via the

[0133] 22 intracerebroventricular, or by intrathecal (cisternal or lumbar) route.

[0134] 23 In various embodiments, there are no more than 10, 9, 8, 7, 6, 5, or 4 administration

[0135] 24 episodes to an individual. In various embodiments, there are no more than three

[0136] 25 administration episodes. In some embodiments, there are no more than two administration

[0137] 26 episodes. For example, in various embodiments there is one administration episode.

[0138] 'Ll In some embodiments, the vector is administered at a multiplicity of infection (MOI)

[0139] 28 selected from the group consisting of 0.5 to 20, 0.5 to 15, 0.5 to 14, 0.5 to 13, 0.5 to 12, 0.5

[0140] 29 to 11, 0.5 to 10, and 1 to 10. The MOI may be 0.25, 0.5 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 1 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24 or 25. Tn some instance administering the

[0141] 2 viral vector at a lower MOI results in increased activity than occurs with administration of

[0142] 3 viral vector at a high MOI. "Multiplicity of Infection" or "MOI" refers to the number of

[0143] 4 virions or virus particles per cell during an infection. A "low MOI" is an MOI below about

[0144] 5 5, below about 2, below about 1, below about 0.1, below about 0.01, below about 0.009,

[0145] 6 below about 0.008, below about 0.007, below about 0.006, below about 0.005, below about

[0146] 7 0.004, below about 0.003, below about 0.002, about 0.001, about 0.0005 or about 0.0001.

[0147] 8 A “high MOI” is an “MOI” above that of 10.

[0148] 9 The term “a” or “an” refers to one or more of that entity, i.e. can refer to a plural

[0149] 10 referent. As such, the terms “a” or “an”, “one or more” and “at least one” are used

[0150] 11 interchangeably herein. In addition, reference to “an element” by the indefinite article “a” or

[0151] 12 “an” does not exclude the possibility that more than one of the elements is present, unless

[0152] 13 the context clearly requires that there is one and only one of the elements.

[0153] 14 The term “about”, unless the context requires otherwise, means ±10% of an

[0154] 15 associated value.

[0155] 16 Numeric ranges are inclusive of the numbers defining the range. The term about is

[0156] 17 used herein to mean plus or minus ten percent (10%) of a value. For example, “about 100”

[0157] 18 refers to any number between 90 and 110. The term “about” a range refers to that range

[0158] 19 minus 10% of its lowest value and plus 10% of its greatest value. Reference to “about” a

[0159] 20 value or parameter herein includes (and describes) embodiments that are directed to that

[0160] 21 value or parameter per se.

[0161] 22 By “polynucleotide cassette” is meant a polynucleotide sequence comprising two or

[0162] 23 more functional polynucleotide sequences, e.g., regulatory elements, translation initiation

[0163] 24 sequences, coding sequences, termination sequences, etc. typically in operable linkage to at

[0164] 25 least one other functional polynucleotide sequence in the polynucleotide cassette. Generally,

[0165] 26 a subject polynucleotide cassette is composed of DNA. The polynucleotide cassettes of the

[0166] 'Ll present disclosure typically comprise a promoter region.

[0167] 28 Furthermore, to the extent that the terms “including”, “includes”, “having”, “has”,

[0168] 29 “with”, or variants thereof are used in either the detailed description and / or the claims, such terms are intended to be inclusive in a manner similar to the term “comprising”. The term “comprising” as used herein is synonymous with “including” or “containing,” and is inclusive or open-ended.

[0169] By “consisting essentially of’, is intended a limitation of the scope of the, for example, composition, method, kit, etc., described to the specified materials that do not materially affect the basic and novel character! stic(s) of the, for example, composition, method, kit, etc. For example, an expression cassette “consisting essentially of’ a coding sequence encoding a polynucleotide operably linked to a promoter and a polyadenylation sequence may include additional sequences, e.g., linker sequences so long as they do not materially affect the transcription or translation of the coding sequence. As another example, a variant or mutant polypeptide “consisting essentially of’ a recited sequence has the amino acid sequence of the recited sequence plus or minus about 10 amino acid residues at the boundaries of the sequence based upon the full length naive polypeptide from which it was derived, e.g. 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 residue less than the recited bounding amino acid residue or 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 residues more than the recited bounding amino acid residue.

[0170] Any reference to “or” herein is intended to encompass “and / or” unless otherwise stated.

[0171] The terms “treat,” “treating”, “treatment,” “ameliorate” or “ameliorating” and other grammatical equivalents as used herein, refer to alleviating, abating or ameliorating a disease or disorder, or symptoms of a disease or disorder, preventing additional symptoms of the disease or disorder, ameliorating or preventing the underlying causes of symptoms, inhibiting a disease or disorder, e.g., arresting the development of a disease or disorder, relieving a disease or disorder, causing regression of a disease or disorder, or stopping the symptoms of a disease or disorder, and are intended to include prophylaxis and prevention. The terms further include achieving a therapeutic benefit and / or a prophylactic benefit. The term “therapeutic benefit” refers to eradication or amelioration of a disease or disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with a disease or disorder such that an improvement is observed in the subject, notwithstanding that, in some embodiments, the subject is still afflicted with a disease or disorder. For prophylactic benefit, the pharmaceutical compositions are administered to a subject at risk of developing a disease or disorder, or to a subject reporting one or more of the physiological symptoms of a disease or disorder, even if a diagnosis of the disease or disorder has not been made.

[0172] The terms “treat,” “treating”, “treatment,” “ameliorate” or “ameliorating” and other grammatical equivalents as used herein may refer to alleviating, abating or ameliorating a lysosome storage-related disease or disorder, or symptoms of lysosome storage-related disease or disorder, preventing additional symptoms of the lysosome storage-related disease or disorder, ameliorating or preventing the underlying causes of symptoms, inhibiting lysosome storage-related disease or disorder, e.g., arresting the development of lysosome storage-related disease or disorder, relieving lysosome storage-related disease or disorder, causing regression of lysosome storage-related disease or disorder, or stopping the symptoms of lysosome storage-related disease or disorder and may include prophylaxis and prevention of lysosome storage-related disease or disorder. The terms further include achieving a therapeutic benefit and / or a prophylactic benefit. The term “therapeutic benefit” of lysosome storage-related disease or disorder refers to eradication or amelioration of the LSD disease or disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with an LSD disease or disorder such that an improvement is observed in the subject, notwithstanding that, in some embodiments, the subject is still afflicted with an LSD disease or disorder. For prophylactic benefit, the pharmaceutical compositions are administered to a subject at risk of developing an LSD disease or disorder, or to a subject reporting one or more of the physiological symptoms of an LSD disease or disorder, even if a diagnosis of the disease or disorder has not been made.

[0173] LSD diseases and disorders include, but are not limited to, spastic paraplegia 11 (SPG11), autosomal recessive hereditary spastic paraplegia (HSP), Charcot-Marie-Tooth (CMT) disease and progressive juvenile-onset amyotrophic lateral sclerosis (ALS).

[0174] Signs and symptoms of an LSD disease or disorder include, but are not limited to, changes in lysosomal number, lysosomal size and lysosomal acidity, upper and lower motor neuron changes, abnormally large abdominal organs, changes to skeletal muscle, coarse 1 facial features, developmental delay, progressive spasticity and weakness of the lower limbs,

[0175] 2 mild intellectual disability, learning difficulties, progressive cognitive decline, peripheral

[0176] 3 neuropathy, pseudobulbar involvement, increased reflexes in the upper limbs, ataxia,

[0177] 4 nystagmus, saccadic pursuit, retinal degeneration, pes cavus, scoliosis, parkinsonism,

[0178] 5 thinning of the corpus callosum, cortical atrophy, and white matter hyerintensities.

[0179] 6 The terms “administer,” “administering”, “administration,” and the like, as used

[0180] 7 herein, can refer to the methods that are used to enable delivery of therapeutics or

[0181] 8 pharmaceutical compositions to the desired site of biological action. These methods include

[0182] 9 injection to a neuron.

[0183] 10 The terms “effective amount”, “therapeutically effective amount” or

[0184] 11 “pharmaceutically effective amount” as used herein, can refer to a sufficient amount of at

[0185] 12 least one pharmaceutical composition or compound being administered which will relieve

[0186] 13 to some extent one or more signs or symptoms of the LSD related disease, LSD disorder or

[0187] 14 LSD related condition being treated. An “effective amount”, “therapeutically effective

[0188] 15 amount” or “pharmaceutically effective amount” of a pharmaceutical composition may be

[0189] 16 administered to a subject in need thereof as a unit dose (as described in further detail

[0190] 17 elsewhere herein). The subject may be a human or non-human mammal.

[0191] 18 The term “pharmaceutically acceptable” as used herein, can refer to a material, such

[0192] 19 as a carrier or diluent, which does not abrogate the biological activity or properties of a

[0193] 20 compound disclosed herein, and is relatively nontoxic (i.e., when the material is

[0194] 21 administered to an individual it does not cause undesirable biological effects nor does it

[0195] 22 interact in a deleterious manner with any of the components of the composition in which it

[0196] 23 is contained).

[0197] 24 The term “pharmaceutical composition,” or simply “composition” as used herein,

[0198] 25 can refer to a biologically active compound, optionally mixed with at least one

[0199] 26 pharmaceutically acceptable chemical component, such as, though not limited to carriers,

[0200] 'Ll stabilizers, diluents, dispersing agents, suspending agents, thickening agents, excipients and

[0201] 28 the like.

[0202] 29 EXAMPLES

[0203] SPG11 is the gene encoding for spatacsin, and is expressed in a broad array of tissues and cells. Mutations in the SPG11 gene can result in a form of autosomal recessive hereditary spastic paraplegia (HSP) as well as Charcot-Marie-Tooth (CMT) disease and progressive juvenile-onset amyotrophic lateral sclerosis (ALS). Examination of the cellular alterations in a mouse model suggests that the loss of spatacsin leads to the accumulation of lipids in lysosomes by perturbing their clearance from these organelles. Brachu J., et al. Loss of spatacsin function alters lysosomal lipid clearance leading to upper and lower motor neuron degeneration. NeurobioL of Dis. Vol. 102: 21-37 (2017). The mechanism is illustrated in FIG. 1.

[0204] Induced pluripotent stem cells (iPSCs) were created from cells of a patient having a SPG11 deficiency. The iPSCs were differentiated into neurons using known techniques.

[0205] Lentivirus vector was prepared encoding the SPG11 gene under the control of EFla promoter (SEQ ID NO: 5). Lentivirus vector was prepared according to known processes using a producer strain (Charles River Laboratory). Lentivirus constructs included a CMV promoter controlling expression of the vector RNA. Lentivirus vector carrying the SPG11 gene were used to infect the neurons derived from patient cells. All LentiViral preps yielded greater than 109Transfecting unit per mL.

[0206] Cells were visualized using LysoTracker Red before and after treatment with lentiviral vector carrying the SPG11 gene. LysoTracker Red is a fluorescent dye specifically designed to label and track acidic organelles in live cells, such as lysosomes. The dye is highly selective for acidic environments, enabling it to accumulate selectively within lysosomes due to their lower internal pH relative to the cytoplasm. Once inside the lysosome, LysoTracker Red becomes protonated and, consequently, trapped within the organelle, preventing its diffusion back into the cytoplasm or other cellular compartments. This selective accumulation allows LysoTracker Red to fluoresce brightly when excited by the appropriate wavelength of light, thereby providing a vivid, real-time visualization of lysosomal morphology and dynamics. As shown in FIG. 2, the diseased neuron fluoresces brightly with LysoTracker Red, and this fluorescence is lost after treatment with the lentiviral vector carrying the SPG11 gene. These studies demonstrate that delivery of the SPG11 gene (including via the EFla promoter) can restore lysosomal dynamics, including in neurons.

[0207] Example 1. IPSC Development

[0208] Whole blood was collected from a patient with an SPG11 disorder. Peripheral blood mnocytes (PBMC) were isolated via Ficoll gradient. The PBMCs were cultured with cytokines (e.g. IL -2) for 3-5 days. Yamanaka factors (OCT4, SOX2, KLF4, c-MYC) using nucleofection. The cells were plated on feeder-free matrix, and the media was switched to reprogramming media. After approximately 2-3 weeks, colonies with iPSC morphology were selected. Markers (OCT4, TRA-1-60, SSEA-4) were confirmed. Chromosomal integrity was confirmed by karyotyping (G-band). Ectoderm, mesoderm and endoderm markers were verified. Validated clones were frozen for storage. Micrographs of representative iPSCs are shown in Fig. 3. G-band karyotyping results not shown.

[0209] Example 2, LysoTracker™ Red Assay

[0210] Lysosomal storage diseases (LSDs) exhibit pathologies including, but not limited to, changes in lysosomal number, lysosomal size and lysosomal acidity. LysoTracker™, a fluorescent dye, selectively accumulates in acidic organelles. LysoTracker™ becomes more intensely retained in dysfunctional lysosomes. Increased LysoTracker™ red accumulation reflects increased lysosomal burden and altered homeostasis. The LysoTracker™ Red Assay is effective in LSDs analysis because it detects changes in lysosomal number, size and acidity.

[0211] Neural stem cells (NSCs) were plated on poly-L-ornithine / laminin-coated surfaces. LysoTracker™ Red was prepared (50-100 nM in warm culture media). Cells were incubated with dye at 37°C for 30 min while protected from light. Cells were washed gently with PBS. Cells were imaged live (Ex / Em 577 / 590 nm) or analyzed by flow cytometry. Unstained controls and treated controls were included. Example 3. Red / Blue Analysis

[0212] NSC’s were stained as described in the LysoTracker™ assay. The cells were also stained with DAPI. Images were obtained. The scale was calibrated using a 150 pm bar. Split channels were used (blue revealing nuclei stained with DAPI), red revealing lysosomes transduced with LysoTracker™ Red). The areas of red signal and blue signal were measured. The ratio of red area / blue area was calculated. The red area / blue area ratio indicates relative transduction. Assays were validated by checking for artifacts and consistent thresholding. Images obtained from one such workflow (8 fields) are shown in Figure 4. The unmasked images are shown in Fig. 4A. The filtered images are shown in Fig. 4B (red Lysotracker area) and 4C (blue DAPI only).

[0213] Example 4, Evaluation of Lentivirus SPG11 on NSCs

[0214] NSCs were developed from a patient exhibiting an LSDs. Cell samples were either untreated (Fig. 5A) or incubated with lentivirus comprising SPG11 at the indicated multiplicity of infection (MOI). Cells were treated with lentivirus at 1 MOI, 2 MOI, 5 MOI or 10 MOI. After incubation, the cells were stained with LysoTracker™ Red and DAPI. The stained cells were evaluated by the Red / Blue Analysis process. Photomicrographs of stained cells treated with 1 MOI lentivirus comprising SPG11 (Fig. 5B), 2 MOI lentivirus comprising SPG11 (Fig. 5C), 5 MOI lentivirus comprising SPG11 (Fig. 5D) or 10 MOI lentivirus comprising SPG11 (Fig. 5E) are shown. Untreated cells from the diseased subject show a substantially higher red / blue ratio than treated cells. The red / blue ratio is substantially reduced at all MOI tested. Treatment with lentivirus comprising SPG11 at an MOI of 1, 2 or 5 was particularly effective, demonstrating red / blue ratios below those observed in cells obtained from a separate subject with an LSDs corrected by CRISPER. Treatment with lentivirus comprising SPG11 at an MOI of 10 resulted in a red / blue ratio comparable to results obtained from cells obtained from a separate subject with an LSDs corrected by CRISPER. The red-blue ratio results from one such experiment are summarized in Fig. 5F. SEQ ID NO:5 and SEQ ID NO:6 were used in various experiments. Example 5. Lentiviral Treatment of SPG11 -Null Mice Trial

[0215] High-titer equine infectious anemia virus (EIAV)-based vectors pseudotyped with the rabies-virus glycoprotein (RV-G) comprising SGP11 are produced. 4 groups of 12 SPG11 null mice / group (allows 6 males and 6 females) for a total of 48 mice are treated with RV-G comprising SGP11 at a predetermined concentration. Body weight is evaluated weekly. Survival is monitored. Blood and tissue samples are collected at intervals and at study completion. Clinical chemistry assays are performed on blood samples from the mice. SPG11 expression is evaluated by qPCR on 6 mice / group (3M / 3F) and 3 tissues / mouse.

[0216] SEQUENCES

[0217] SEQ ID NO : 1 SPG11 Amino Acid Sequence ( 2443 aa )

[0218] MAAEEGVASAASAGGSWGTAAMGRVLPMLLVPVPAEAMGQLGSRAQLRTQPEALGSLTAAGSLQVL SLTPGSRGGGRCCLEGPFWHFLWEDSRNSSTPTEKPKLLALGENYELLIYEFNLKDGRCDAT ILYS CSREALQKLIDDQDI SI SLLSLRILSFHNNTSLLFINKCVILHI I FPERDAAIRVLNCFTLPLPAQ AVDMI IDTQLCRGILFVLSSLGWIY IFDVVDGTYVAHVDLALHKEDMCNEQQQEPAKI SS FTSLKV SQDLDVAVIVSSSNSAVALNLNLYFRQHPGHLLCERILEDLPIQGPKGVDEDDPVNSAYNMKLAKF S FQIDRSWKAQLSSLNETIKNSKLEVSCCAPWFQDILHLESPESGNHSTSVQSWAFIPQDIMHGQY NVLQKDHAKTSDPGRSWKIMHISEQEEPIELKCVSVTGFTALFTWEVERMGYTITLWDLETQGMQC FSLGTKCIPVDSSGDQQLCFVLTENGLSLILFGLTQEEFLNRLMIHGSASTVDTLCHLNGWGRCSI PIHALEAGIENRQLDTVNFFLKSKENLFNPSSKSSVSDQFDHLSSHLYLRNVEELI PALDLLCSAI RESYSEPQSKHFSEQLLNLTLSFLNNQIKELFIHTEELDEHLQKGVNILTSY INELRT FMIKFPWK LTDAIDEYDVHENVPKVKESNIWKKLS FEEVIASAILNNKI PEAQTFFRIDSHSAQKLEELIGIGL NL V FDNL KKNN I KE AS E LL KNMG FD VKGQLL KI C F YT TNKN I RD FL VE I L KE KN Y F S E KE KRT I D F VHQVEKLYLGHFQENMQIQSFPRYWIKEQDFFKHKSVLDSFLKYDCKDEFNKQDHRIVLNWALWWD QLTQESILLPRISPEEYKSYSPEALWRYLTARHDWLNIILWIGEFQTQHSYASLQQNKWPLLTVDV INQNTSCNNYMRNEILDKLARNGVFLASELEDFECFLLRLSRIGGVIQDTLPVQNYKTKEGWDFHS QFILYCLEHSLQHLLYVYLDCYKLSPENCPFLEKKELHEAHPWFE FLVQCRQVASNLTDPKLIFQA SLANAQILI PTNQASVSSMLLEGHTLLALATTMYSPGGVSQVVQNEENENCLKKVDPQLLKMALTP YPKLKTALFPQCTPPSVLPSDIT IYHLIQSLSPFDPSRLFGWQSANTLAIGDAWSHLPHFSSPDLV NKYAIVERLNFAYYLHNGRPS FAFGTFLVQELIKSKTPKQLIQQVGNEAYVIGLSS FHIPSIGAAC VCFLELLGLDSLKLRVDMKVANI ILSYKCRNEDAQYS FIRESVAEKLSKLADGEKTTTEELLVLLE EGTWNSIQQQE IKRLSSESSSQWALWQFCRLHNMKLSI SYLRECAKANDWLQFIIHSQLHNYHPA EVKSLIQYFSPVIQDHLRLAFENLPSVPTSKMDSDQVCNKCPQELQGSKQEMTDLFEILLQCSEEP DSWHWLLVEAVKQQAPILSVLASCLQGASAI SCLCVWIITSVEDNVATEAMGHIQDSTEDHTWNLE DLSVIWRTLLTRQKSKTLIRGFQLFFKDSPLLLVMEMYELCMFFRNYKEAEAKLLE FQKSLETLNT

[0219] AATKVHPVI PAMWLEDQVCFLLKLMLQQCKTQYELGKLLQLFVEREHLFSDGPDVKKLCILCQILK DTS IAINHT IITSYS IENLQHECRS ILERLQTDGQFALARRVAELAELPVDNLVIKEITQEMQTLK HIEQWSLKQARIDFWKKCHENFKKNSI SSKAASSFFSTQAHVACEHPTGWSSMEERHLLLTLAGHW LAQEDVVPLDKLEELEKQIWLCRITQHTLGRNQEETEPRFSRQISTSGELSFDSLASE FS FSKLAA LNTSKYLELNSLPSKETCENRLDWKEQESLNFLIGRLLDDGCVHEASRVCRY FHFYNPDVALVLHC RALASGEASMEDLHPEIHALLQSAELLEEEAPDIPLRRVHSTSSLDSQKFVTVPSSNEVVTNLEVL TSKCLHGKNYCRQVLCLYDLAKELGCSYTDVAAQDGEAMLRKILASQQPDRCKRAQAFISTQGLKP DTVAELVAEEVTRELLTSSQGTGHKQMFNPTEESQTFLQLTTLCQDRTLVGMKLLDKI SSVPHGEL 1 SCTTELLILAHHCFTLTCHMEGI IRVLQAAHMLTDNHLAPSEEYGLVVRLLTGIGRYNEMTY IFDL

[0220] 2 LHKKHYFEVLMRKKLDPSGTLKTALLDYIKRCRPGDSEKHNMIALCFSMCRE IGENHEAAARIQLK

[0221] 3 LIESQPWEDSLKDGHQLKQLLLKALTLMLDAAESYAKDSCVRQAQHCQRLTKLITLQIHFLNTGQN

[0222] 4 TMLINLGRHKLMDCILALPRFYQAS IVAEAYDFVPDWAE ILYQQVILKGDFNYLEE FKQQRLLKSS

[0223] 5 I FEEI SKKYKQHQPTDMVMENLKKLLTYCEDVYLYYKLAYEHKFYEIVNVLLKDPQTGCCLKDMLA

[0224] 6 G

[0225] 7 SEQ ID NO : 2 - SPG11 cDNA

[0226] 8 ATGGCTGCAGAGGAAGGGGTCGCGAGTGCTGCTTCCGCCGGCGGTAGCTGGGGCACCGCGGCCATG

[0227] 9 GGGCGGGTTCTACCGATGCTGTTGGTGCCAGTCCCCGCCGAGGCGATGGGGCAGCTCGGCTCCCGG

[0228] 10 GCGCAGCTGCGCACACAGCCGGAGGCTCTGGGGAGCCTGACGGCTGCGGGCAGCCTCCAAGTGCTT

[0229] 11 TCTTTGACGCCTGGCAGCCGGGGCGGGGGTCGCTGCTGCCTGGAGGGCCCCTTCTGGCACTTTCTA

[0230] 12 TGGGAGGATTCTCGTAACAGCAGCACACCAACTGAAAAGCCCAAACTGCTCGCTCTTGGTGAAAAT

[0231] 13 TATGAACTGCTTATCTATGAATTTAATTTGAAAGATGGAAGATGTGATGCAACCATTTTGTATAGC

[0232] 14 TGTAGTAGGGAGGCATTGCAAAAGCTCATTGACGATCAAGATATCAGTATTTCCTTATTGTCTTTG

[0233] 15 AGAATCCTGTCATTTCACAATAACACATCATTACTGTTCATCAACAAATGTGTCATCCTACATATT

[0234] 16 ATATTTCCTGAAAGAGATGCTGCAATTAGAGTACTCAACTGTTTCACACTTCCCTTGCCTGCACAG

[0235] 17 GCAGTGGACATGATTATTGACACGCAGCTCTGCAGAGGAATTCTTTTTGTTTTGAGTAGTTTAGGC

[0236] 18 TGGATCTACATTTTTGATGTTGTGGATGGTACATATGTAGCTCATGTGGATTTAGCACTTCACAAA

[0237] 19 GAAGACATGTGTAATGAGCAGCAACAGGAGCCAGCCAAGATTTCTTCATTTACTTCACTGAAAGTT

[0238] 20 TCTCAAGACCTCGATGTTGCAGTGATTGTCAGCTCCTCCAACTCCGCAGTTGCTCTTAACTTAAAT

[0239] 21 TTGTATTTCAGGCAACACCCAGGACACCTACTGTGTGAAAGAATACTAGAAGATCTTCCTATTCAA

[0240] 22 GGACCTAAGGGCGTAGATGAAGATGATCCTGTTAACTCTGCCTACAACATGAAACTGGCCAAGTTT

[0241] 23 TCCTTCCAAATTGATAGGTCTTGGAAAGCCCAGCTATCATCATTGAATGAAACAATAAAGAACTCC

[0242] 24 AAACTGGAGGTTTCCTGTTGTGCTCCATGGTTCCAGGATATTTTGCATTTGGAGTCACCTGAATCT

[0243] 25 GGTAACCACAGTACAAGTGTGCAGAGCTGGGCCTTCATTCCACAGGACATAATGCATGGGCAATAT

[0244] 26 AAT GT T C T AC AGAAAGATC AT GC C AAG AC CAGT GAT C CAGG AAGAT C AT GGAAAAT AAT GC AC AT C

[0245] 'Ll AGTGAACAAGAGGAACCCATAGAGCTTAAATGTGTGTCTGTGACAGGATTCACTGCACTGTTTACT

[0246] 28 TGGGAAGTGGAAAGGATGGGCTATACCATTACCCTCTGGGATTTGGAGACCCAGGGCATGCAGTGT

[0247] 29 TTTTCCCTTGGCACAAAGTGTATTCCTGTAGACAGTAGTGGAGACCAGCAGCTGTGCTTTGTTTTG

[0248] 30 ACAGAGAATGGACTCTCTCTGATTTTGTTTGGTTTGACTCAAGAAGAGTTTTTAAACAGACTCATG

[0249] 31 ATCCATGGAAGTGCCAGCACTGTGGACACTCTTTGTCATCTCAATGGCTGGGGAAGGTGCTCAATT

[0250] 32 CCCATACATGCACTAGAGGCCGGGATAGAAAATCGTCAGCTGGACACAGTAAATTTCTTTTTGAAG

[0251] 33 AGC AAGG AAAAT C T T T T T AAT CC AT CC T C AAAATC T T CT GT AT CT GAT CAGT T T GAT C AC T T GT C A

[0252] 34 TCCCATTTATATTTAAGAAATGTGGAAGAGCTGATACCAGCATTGGATTTACTTTGCTCGGCAATT AGAGAAAGTTATTCTGAACCCCAAAGCAAACACTTTTCAGAACAATTGCTTAATCTTACACTGTCT

[0253] TTCCTTAACAACCAAATAAAGGAGCTTTTCATTCACACTGAAGAACTAGATGAACATCTGCAAAAA

[0254] GGAGTGAACATTTTGACTAGCTACATTAATGAACTTCGAACCTTCATGATAAAGTTTCCTTGGAAG

[0255] CTAACAGATGCTATAGATGAATATGATGTACATGAAAATGTCCCCAAAGTAAAGGAGAGCAATATA

[0256] TGGAAGAAACTCAGCTTTGAGGAAGTTATTGCCAGCGCCATTTTAAACAACAAAATACCAGAGGCA

[0257] CAGACTTTCTTCAGGATTGATAGTCATTCTGCTCAAAAACTTGAGGAGCTTATTGGCATAGGCCTA

[0258] AATTTGGTCTTTGACAATTTAAAAAAGAACAATATAAAGGAAGCCTCTGAACTTTTGAAGAATATG

[0259] GGGTTTGATGTAAAAGGCCAATTGCTCAAGATCTGCTTCTATACAACTAATAAAAATATACGTGAC

[0260] TTTTTGGTTGAAATTTTAAAAGAAAAAAATTATTTTTCTGAAAAAGAGAAAAGAACTATAGACTTC

[0261] GTGCATCAAGTTGAGAAGCTTTATTTGGGACATTTCCAAGAAAATATGCAAATCCAGTCATTTCCC

[0262] AGGTACTGGATAAAGGAACAAGATTTTTTCAAGCACAAGTCTGTTTTGGACTCATTCCTGAAATAT

[0263] GATTGTAAAGATGAATTTAACAAACAGGACCATAGAATTGTGTTAAATTGGGCTCTGTGGTGGGAT

[0264] CAACTAACACAAGAATCCATCCTTCTCCCCAGGATAAGTCCAGAAGAATACAAATCATATTCCCCT

[0265] GAAGCCCTCTGGAGATACCTCACAGCTCGCCATGATTGGTTAAACATTATCTTATGGATTGGAGAA

[0266] TTTCAAACCCAGCATAGTTATGCTTCACTTCAGCAGAACAAATGGCCCCTTCTGACTGTTGATGTT

[0267] ATTAACCAGAATACTTCCTGTAACAACTACATGAGGAATGAAATTTTAGATAAGCTGGCCAGGAAT

[0268] GGGGTTTTTTTGGCATCTGAACTGGAAGACTTTGAATGCTTCCTCCTAAGACTGAGCCGTATTGGA

[0269] GGTGTAATACAGGATACCCTCCCTGTTCAAAACTACAAGACCAAAGAAGGTTGGGATTTCCATTCT

[0270] CAATTCATTCTCTATTGTTTGGAGCACAGTCTGCAGCATCTTCTTTATGTCTACCTTGACTGTTAC

[0271] AAACTTAGTCCTGAAAATTGTCCCTTTTTGGAAAAAAAAGAGTTACATGAAGCACACCCTTGGTTT

[0272] GAATTTTTAGTTCAGTGTCGACAAGTTGCCAGTAACTTAACAGATCCCAAACTGATCTTCCAGGCT

[0273] AGCCTTGCAAATGCTCAGATTTTGATTCCCACCAATCAGGCCAGTGTAAGCAGTATGCTATTGGAA

[0274] GGACATACCCTCCTGGCCCTTGCTACTACAATGTATTCTCCTGGGGGTGTCAGTCAGGTTGTTCAG

[0275] AATGAAGAAAATGAAAACTGTTTGAAGAAAGTGGATCCCCAGCTATTGAAGATGGCATTAACTCCT

[0276] TACCCCAAGCTAAAAACTGCTCTCTTCCCACAGTGCACTCCTCCTAGTGTCCTGCCATCTGATATT

[0277] ACAATCTACCACCTTATTCAGTCATTATCACCCTTTGATCCTAGCAGATTGTTTGGCTGGCAGTCT

[0278] GCTAACACACTAGCTATAGGAGATGCATGGAGTCATCTCCCACATTTCTCTAGCCCTGACCTGGTT

[0279] AATAAATATGCTATAGTGGAACGTCTGAATTTTGCTTATTATTTACATAATGGGCGGCCATCATTT

[0280] GCATTTGGTACTTTTCTGGTCCAGGAATTAATCAAGAGCAAGACTCCCAAGCAGCTGATCCAGCAA

[0281] GTAGGCAATGAAGCCTATGTTATAGGGCTCTCCTCCTTCCACATACCTTCAATAGGAGCTGCATGT

[0282] GTTTGTTTCTTAGAATTGCTTGGCCTTGACAGCCTCAAGCTCAGAGTTGATATGAAAGTGGCCAAT

[0283] ATAATTTTGAGCTACAAGTGCAGAAATGAAGATGCTCAGTACAGCTTTATCAGAGAGTCTGTAGCC

[0284] GAAAAACTATCTAAACTAGCTGATGGTGAAAAGACAACCACAGAAGAATTGCTTGTTCTCTTAGAA

[0285] GAAGGTACATGGAACAGCATTCAGCAACAGGAAATAAAGAGGTTATCCAGTGAATCTAGCAGCCAA

[0286] TGGGCATTAGTGGTGCAGTTCTGCAGGCTACACAATATGAAACTAAGCATATCTTACCTTAGAGAA TGTGCCAAAGCAAATGATTGGCTGCAGTTCATTATTCACAGCCAACTCCACAACTACCACCCAGCA GAGGTGAAATCCCTTATCCAGTACTTCAGCCCAGTCATTCAAGACCACTTAAGGCTGGCTTTTGAG AACTTGCCCTCAGTGCCCACCTCCAAAATGGACAGCGATCAAGTCTGCAATAAGTGCCCCCAGGAA CTTCAAGGAAGCAAACAAGAGATGACCGATTTATTTGAAATTCTGCTCCAATGCTCAGAGGAGCCA GACTCCTGGCACTGGCTTCTGGTTGAAGCAGTGAAACAACAGGCCCCTATCCTCAGTGTTCTGGCC TCATGTCTCCAGGGTGCCAGTGCCATTTCTTGTCTCTGTGTTTGGATCATCACTTCTGTGGAGGAC AATGTTGCAACTGAAGCAATGGGACACATTCAGGACTCAACAGAGGACCATACCTGGAACCTTGAG GAT CT T T C AGT CAT C T GGAGAAC AT T AT T AAC AAG AC AAAAGAGC AAAAC T C T C AT C AGAGGT T T C CAGCTTTTCTTTAAGGATTCCCCGTTACTACTGGTGATGGAGATGTATGAACTGTGTATGTTCTTC AGGAATTATAAAGAAGCTGAAGCTAAACTTCTGGAGTTTCAGAAGAGCCTTGAAACGCTTAACACA GCAGCCACAAAGGTCCACCCTGTCATCCCTGCCATGTGGCTGGAGGATCAGGTGTGTTTCCTTTTG AAGCTTATGCTACAGCAGTGTAAGACCCAGTATGAGCTGGGGAAGCTTTTACAGCTCTTTGTTGAA AGAGAGCATCTCTTCTCTGATGGTCCAGATGTGAAAAAGCTTTGCATCCTTTGCCAGATTTTGAAG GATACATCCATAGCCATTAATCATACAATTATTACCAGCTACAGCATTGAGAATCTTCAGCATGAA TGTAGATCTATTTTGGAAAGACTGCAGACAGATGGACAATTCGCTTTGGCCAGGAGGGTAGCAGAA TTAGCTGAGTTACCTGTGGACAACTTGGTTATTAAAGAGATAACACAGGAAATGCAGACCCTAAAA C AC AT T G AAC AGT GGT C AC T AAAAC AAGCAAGAAT T G AC T T CT GG AAAAAAT GC CAT G AG AAT T T T AAGAAAAATTCAATTTCAAGCAAAGCAGCTTCTTCCTTTTTCTCAACCCAGGCCCATGTGGCATGT GAGCACCCAACTGGATGGAGCAGCATGGAGGAGCGCCATCTGCTGCTCACCTTGGCAGGGCACTGG CTTGCCCAGGAGGACGTGGTGCCCTTGGATAAGCTGGAGGAGCTGGAGAAGCAGATCTGGCTGTGC CGCATCACCCAGCACACTCTTGGAAGAAATCAGGAGGAAACAGAGCCCAGATTTTCTCGACAGATC TCAACTAGTGGTGAACTTTCCTTTGATAGTTTAGCCAGTGAGTTTTCCTTCTCCAAGTTGGCTGCT C T G AAC AC AT C AAAAT ACT T AGAAC TT AACAGC CT T C CAT C C AAAGAG AC AT GC GAGAAT AG AT T G GATTGGAAAGAGCAGGAGTCACTAAACTTTTTGATTGGGCGCCTACTGGATGATGGCTGTGTGCAT GAAGCAAGTAGAGTATGCCGGTATTTTCATTTTTATAATCCAGATGTCGCCTTGGTATTGCACTGC AGAGCACTGGCCTCAGGGGAAGCTAGTATGGAGGATCTGCACCCAGAGATCCATGCTCTCCTACAA AGTGCTGAGCTGCTTGAGGAAGAAGCACCCGACATTCCCCTAAGGAGAGTCCACAGCACTTCAAGT CTGGATAGTCAGAAGTTTGTGACAGTGCCCTCCAGTAATGAAGTGGTAACTAACCTGGAAGTGCTG ACAAGCAAATGCCTCCATGGGAAGAACTACTGTCGACAGGTCCTCTGTCTGTATGATCTTGCCAAG GAGTTGGGCTGTTCCTACACAGATGTTGCTGCTCAGGATGGTGAAGCCATGCTCCGGAAAATCTTG GCCTCTCAGCAGCCTGACCGATGCAAACGAGCCCAGGCCTTCATCAGCACACAGGGCCTTAAGCCA GATACTGTGGCTGAACTCGTGGCAGAAGAGGTGACACGGGAGCTGCTTACTTCATCACAGGGAACA GGACATAAGCAGATGTTCAACCCAACAGAGGAAAGCCAGACATTTCTTCAGCTGACCACTCTGTGT CAAGACCGCACATTGGTAGGCATGAAGTTGTTGGATAAGATTTCCTCCGTTCCCCATGGGGAACTG TCTTGCACCACAGAGCTCCTGATCCTGGCCCATCATTGCTTCACCCTGACGTGCCACATGGAGGGC 1 ATCATCCGAGTCCTACAGGCCGCCCACATGCTCACAGATAACCACCTGGCCCCCAGTGAGGAGTAT

[0287] 2 GGGCTGGTGGTACGGCTCCTCACTGGCATTGGAAGGTACAACGAGATGACATACATATTTGATTTG

[0288] 3 CTGCATAAAAAGCACTACTTTGAAGTGCTAATGAGGAAGAAGTTGGATCCGAGTGGTACCCTGAAA

[0289] 4 ACAGCCCTGCTGGACTACATCAAACGCTGCCGTCCTGGAGACAGTGAAAAGCACAATATGATTGCC

[0290] 5 CTGTGCTTCAGCATGTGCCGGGAGATTGGCGAGAACCACGAGGCAGCTGCCCGCATCCAACTGAAA

[0291] 6 TTGATTGAGTCTCAGCCCTGGGAGGACAGCCTCAAGGATGGGCACCAGCTGAAACAACTGCTGCTG

[0292] 7 AAGGCCCTGACTCTGATGTTGGATGCAGCAGAGAGTTATGCCAAGGACTCCTGTGTGCGACAGGCC

[0293] 8 CAGCACTGTCAGCGGCTCACCAAGTTGATAACTCTGCAGATTCACTTTCTGAACACTGGCCAGAAC

[0294] 9 ACAATGCTCATCAACTTGGGCCGCCACAAGCTGATGGACTGTATTCTGGCCCTACCTCGGTTCTAC

[0295] 10 CAGGCTTCTATTGTGGCTGAGGCCTACGATTTTGTTCCAGATTGGGCTGAAATTTTATACCAGCAA

[0296] 11 GTGATTCTTAAAGGAGACTTTAATTACTTGGAAGAATTTAAGCAGCAAAGGTTATTAAAGTCCAGT

[0297] 12 ATATTTGAAGAGATTTCCAAAAAATATAAACAACATCAGCCTACTGACATGGTCATGGAAAACCTG

[0298] 13 AAGAAATTACTCACATATTGTGAAGATGTTTACCTGTATTACAAGTTGGCATACGAACACAAGTTT

[0299] 14 TATGAAATTGTAAATGTGCTTCTGAAGGACCCTCAGACAGGTTGCTGTCTAAAGGACATGCTAGCA

[0300] 15 GGTTAG

[0301] 16 SEQ ID NO : 3 - Codon optimized SPG11 cDNA

[0302] 17 ATGGCTGCAGAGGAAGGGGTCGCGAGTGCTGCTTCCGCCGGCGGTAGCTGGGGCACCGCGGCCATG

[0303] 18 GGGCGGGTTCTACCGATGCTGTTGGTGCCAGTCCCCGCCGAGGCGATGGGGCAGCTCGGCTCCCGG

[0304] 19 GCGCAGCTGCGCACACAGCCGGAGGCTCTGGGGAGCCTGACGGCTGCGGGCAGCCTCCAAGTGCTT

[0305] 20 TCTTTGACGCCTGGCAGCCGGGGCGGGGGTCGCTGCTGCCTGGAGGGCCCCTTCTGGCACTTTCTA

[0306] 21 TGGGAGGATTCTCGTAACAGCAGCACACCAACTGAAAAGCCCAAACTGCTCGCTCTTGGTGAAAAT

[0307] 22 TATGAACTGCTTATCTATGAATTTAATTTGAAAGATGGAAGATGTGATGCAACCATTTTGTATAGC

[0308] 23 TGTAGTAGGGAGGCATTGCAAAAGCTCATTGACGATCAAGATATCAGTATTTCCTTATTGTCTTTG

[0309] 24 AGAATCCTGTCATTTCACAATAACACATCATTACTGTTCATCAACAAATGTGTCATCCTACATATT

[0310] 25 ATATTTCCTGAAAGAGATGCTGCAATTAGAGTACTCAACTGTTTCACACTTCCCTTGCCTGCACAG

[0311] 26 GCAGTGGACATGATTATTGACACGCAGCTCTGCAGAGGAATTCTTTTTGTTTTGAGTAGTTTAGGC

[0312] 'Ll TGGATCTACATTTTTGATGTTGTGGATGGTACATATGTAGCTCATGTGGATTTAGCACTTCACAAA

[0313] 28 GAAGACATGTGTAATGAGCAGCAACAGGAGCCAGCCAAGATTTCTTCATTTACTTCACTGAAAGTT

[0314] 29 TCTCAAGACCTCGATGTTGCAGTGATTGTCAGCTCCTCCAACTCCGCAGTTGCTCTTAACTTAAAT

[0315] 30 TTGTATTTCAGGCAACACCCAGGACACCTACTGTGTGAAAGAATACTAGAAGATCTTCCTATTCAA

[0316] 31 GGACCTAAGGGCGTAGATGAAGATGATCCTGTTAACTCTGCCTACAACATGAAACTGGCCAAGTTT

[0317] 32 TCCTTCCAAATTGATAGGTCTTGGAAAGCCCAGCTATCATCATTGAATGAAACAATAAAGAACTCC

[0318] 33 AAACTGGAGGTTTCCTGTTGTGCTCCATGGTTCCAGGATATTTTGCATTTGGAGTCACCTGAATCT

[0319] 34 GGTAACCACAGTACAAGTGTGCAGAGCTGGGCCTTCATTCCACAGGACATAATGCATGGGCAATAT AAT GT T C T AC AGAAAGATC AT GC C AAG AC CAGT GAT C CAGG AAGAT C AT GGAAAAT AAT GC AC AT C AGTGAACAAGAGGAACCCATAGAGCTTAAATGTGTGTCTGTGACAGGATTCACTGCACTGTTTACT TGGGAAGTGGAAAGGATGGGCTATACCATTACCCTCTGGGATTTGGAGACCCAGGGCATGCAGTGT TTTTCCCTTGGCACAAAGTGTATTCCTGTAGACAGTAGTGGAGACCAGCAGCTGTGCTTTGTTTTG ACAGAGAATGGACTCTCTCTGATTTTGTTTGGTTTGACTCAAGAAGAGTTTTTAAACAGACTCATG ATCCATGGAAGTGCCAGCACTGTGGACACTCTTTGTCATCTCAATGGCTGGGGAAGGTGCTCAATT CCCATACATGCACTAGAGGCCGGGATAGAAAATCGTCAGCTGGACACAGTAAATTTCTTTTTGAAG AGC AAGG AAAAT C T T T T T AAT CC AT CC T C AAAATC T T CT GT AT CT GAT CAGT T T GAT C AC T T GT C A TCCCATTTATATTTAAGAAATGTGGAAGAGCTGATACCAGCATTGGATTTACTTTGCTCGGCAATT AGAGAAAGTTATTCTGAACCCCAAAGCAAACACTTTTCAGAACAATTGCTTAATCTTACACTGTCT TTCCTTAACAACCAAATAAAGGAGCTTTTCATTCACACTGAAGAACTAGATGAACATCTGCAAAAA GGAGTGAACATTTTGACTAGCTACATTAATGAACTTCGAACCTTCATGATAAAGTTTCCTTGGAAG CTAACAGATGCTATAGATGAATATGATGTACATGAAAATGTCCCCAAAGTAAAGGAGAGCAATATA TGGAAGAAACTCAGCTTTGAGGAAGTTATTGCCAGCGCCATTTTAAACAACAAAATACCAGAGGCA CAGACTTTCTTCAGGATTGATAGTCATTCTGCTCAAAAACTTGAGGAGCTTATTGGCATAGGCCTA AATTTGGTCTTTGACAATTTAAAAAAGAACAATATAAAGGAAGCCTCTGAACTTTTGAAGAATATG GGGTTTGATGTAAAAGGCCAATTGCTCAAGATCTGCTTCTATACAACTAATAAAAATATACGTGAC TTTTTGGTTGAAATTTTAAAAGAAAAAAATTATTTTTCTGAAAAAGAGAAAAGAACTATAGACTTC GTGCATCAAGTTGAGAAGCTTTATTTGGGACATTTCCAAGAAAATATGCAAATCCAGTCATTTCCC AGGTACTGGATAAAGGAACAAGATTTTTTCAAGCACAAGTCTGTTTTGGACTCATTCCTGAAATAT GATTGTAAAGATGAATTTAACAAACAGGACCATAGAATTGTGTTAAATTGGGCTCTGTGGTGGGAT CAACTAACACAAGAATCCATCCTTCTCCCCAGGATAAGTCCAGAAGAATACAAATCATATTCCCCT GAAGCCCTCTGGAGATACCTCACAGCTCGCCATGATTGGTTAAACATTATCTTATGGATTGGAGAA TTTCAAACCCAGCATAGTTATGCTTCACTTCAGCAGAACAAATGGCCCCTTCTGACTGTTGATGTT ATTAACCAGAATACTTCCTGTAACAACTACATGAGGAATGAAATTTTAGATAAGCTGGCCAGGAAT GGGGTTTTTTTGGCATCTGAACTGGAAGACTTTGAATGCTTCCTCCTAAGACTGAGCCGTATTGGA GGTGTAATACAGGATACCCTCCCTGTTCAAAACTACAAGACCAAAGAAGGTTGGGATTTCCATTCT CAATTCATTCTCTATTGTTTGGAGCACAGTCTGCAGCATCTTCTTTATGTCTACCTTGACTGTTAC AAACTTAGTCCTGAAAATTGTCCCTTTTTGGAAAAAAAAGAGTTACATGAAGCACACCCTTGGTTT GAATTTTTAGTTCAGTGTCGACAAGTTGCCAGTAACTTAACAGATCCCAAACTGATCTTCCAGGCT AGCCTTGCAAATGCTCAGATTTTGATTCCCACCAATCAGGCCAGTGTAAGCAGTATGCTATTGGAA GGACATACCCTCCTGGCCCTTGCTACTACAATGTATTCTCCTGGGGGTGTCAGTCAGGTTGTTCAG AATGAAGAAAATGAAAACTGTTTGAAGAAAGTGGATCCCCAGCTATTGAAGATGGCATTAACTCCT TACCCCAAGCTAAAAACTGCTCTCTTCCCACAGTGCACTCCTCCTAGTGTCCTGCCATCTGATATT ACAATCTACCACCTTATTCAGTCATTATCACCCTTTGATCCTAGCAGATTGTTTGGCTGGCAGTCT GCTAACACACTAGCTATAGGAGATGCATGGAGTCATCTCCCACATTTCTCTAGCCCTGACCTGGTT AATAAATATGCTATAGTGGAACGTCTGAATTTTGCTTATTATTTACATAATGGGCGGCCATCATTT GCATTTGGTACTTTTCTGGTCCAGGAATTAATCAAGAGCAAGACTCCCAAGCAGCTGATCCAGCAA GTAGGCAATGAAGCCTATGTTATAGGGCTCTCCTCCTTCCACATACCTTCAATAGGAGCTGCATGT GTTTGTTTCTTAGAATTGCTTGGCCTTGACAGCCTCAAGCTCAGAGTTGATATGAAAGTGGCCAAT ATAATTTTGAGCTACAAGTGCAGAAATGAAGATGCTCAGTACAGCTTTATCAGAGAGTCTGTAGCC GAAAAACTATCTAAACTAGCTGATGGTGAAAAGACAACCACAGAAGAATTGCTTGTTCTCTTAGAA GAAGGTACATGGAACAGCATTCAGCAACAGGAAATAAAGAGGTTATCCAGTGAATCTAGCAGCCAA TGGGCATTAGTGGTGCAGTTCTGCAGGCTACACAATATGAAACTAAGCATATCTTACCTTAGAGAA TGTGCCAAAGCAAATGATTGGCTGCAGTTCATTATTCACAGCCAACTCCACAACTACCACCCAGCA GAGGTGAAATCCCTTATCCAGTACTTCAGCCCAGTCATTCAAGACCACTTAAGGCTGGCTTTTGAG AACTTGCCCTCAGTGCCCACCTCCAAAATGGACAGCGATCAAGTCTGCAATAAGTGCCCCCAGGAA CTTCAAGGAAGCAAACAAGAGATGACCGATTTATTTGAAATTCTGCTCCAATGCTCAGAGGAGCCA GACTCCTGGCACTGGCTTCTGGTTGAAGCAGTGAAACAACAGGCCCCTATCCTCAGTGTTCTGGCC TCATGTCTCCAGGGTGCCAGTGCCATTTCTTGTCTCTGTGTTTGGATCATCACTTCTGTGGAGGAC AATGTTGCAACTGAAGCAATGGGACACATTCAGGACTCAACAGAGGACCATACCTGGAACCTTGAG GAT CT T T C AGT CAT C T GGAGAAC AT T AT T AAC AAG AC AAAAGAGC AAAAC T C T C AT C AGAGGT T T C CAGCTTTTCTTTAAGGATTCCCCGTTACTACTGGTGATGGAGATGTATGAACTGTGTATGTTCTTC AGGAATTATAAAGAAGCTGAAGCTAAACTTCTGGAGTTTCAGAAGAGCCTTGAAACGCTTAACACA GCAGCCACAAAGGTCCACCCTGTCATCCCTGCCATGTGGCTGGAGGATCAGGTGTGTTTCCTTTTG AAGCTTATGCTACAGCAGTGTAAGACCCAGTATGAGCTGGGGAAGCTTTTACAGCTCTTTGTTGAA AGAGAGCATCTCTTCTCTGATGGTCCAGATGTGAAAAAGCTTTGCATCCTTTGCCAGATTTTGAAG GATACATCCATAGCCATTAATCATACAATTATTACCAGCTACAGCATTGAGAATCTTCAGCATGAA TGTAGATCTATTTTGGAAAGACTGCAGACAGATGGACAATTCGCTTTGGCCAGGAGGGTAGCAGAA TTAGCTGAGTTACCTGTGGACAACTTGGTTATTAAAGAGATAACACAGGAAATGCAGACCCTAAAA C AC AT T G AAC AGT GGT C AC T AAAAC AAGCAAGAAT T G AC T T CT GG AAAAAAT GC CAT G AG AAT T T T AAGAAAAATTCAATTTCAAGCAAAGCAGCTTCTTCCTTTTTCTCAACCCAGGCCCATGTGGCATGT GAGCACCCAACTGGATGGAGCAGCATGGAGGAGCGCCATCTGCTGCTCACCTTGGCAGGGCACTGG CTTGCCCAGGAGGACGTGGTGCCCTTGGATAAGCTGGAGGAGCTGGAGAAGCAGATCTGGCTGTGC CGCATCACCCAGCACACTCTTGGAAGAAATCAGGAGGAAACAGAGCCCAGATTTTCTCGACAGATC TCAACTAGTGGTGAACTTTCCTTTGATAGTTTAGCCAGTGAGTTTTCCTTCTCCAAGTTGGCTGCT C T G AAC AC AT C AAAAT ACT T AGAAC TT AACAGC CT T C CAT C C AAAGAG AC AT GC GAGAAT AG AT T G GATTGGAAAGAGCAGGAGTCACTAAACTTTTTGATTGGGCGCCTACTGGATGATGGCTGTGTGCAT GAAGCAAGTAGAGTATGCCGGTATTTTCATTTTTATAATCCAGATGTCGCCTTGGTATTGCACTGC AGAGCACTGGCCTCAGGGGAAGCTAGTATGGAGGATCTGCACCCAGAGATCCATGCTCTCCTACAA 1 AGTGCTGAGCTGCTTGAGGAAGAAGCACCCGACATTCCCCTAAGGAGAGTCCACAGCACTTCAAGT

[0320] 2 CTGGATAGTCAGAAGTTTGTGACAGTGCCCTCCAGTAATGAAGTGGTAACTAACCTGGAAGTGCTG

[0321] 3 ACAAGCAAATGCCTCCATGGGAAGAACTACTGTCGACAGGTCCTCTGTCTGTATGATCTTGCCAAG

[0322] 4 GAGTTGGGCTGTTCCTACACAGATGTTGCTGCTCAGGATGGTGAAGCCATGCTCCGGAAAATCTTG

[0323] 5 GCCTCTCAGCAGCCTGACCGATGCAAACGAGCCCAGGCCTTCATCAGCACACAGGGCCTTAAGCCA

[0324] 6 GATACTGTGGCTGAACTCGTGGCAGAAGAGGTGACACGGGAGCTGCTTACTTCATCACAGGGAACA

[0325] 7 GGACATAAGCAGATGTTCAACCCAACAGAGGAAAGCCAGACATTTCTTCAGCTGACCACTCTGTGT

[0326] 8 CAAGACCGCACATTGGTAGGCATGAAGTTGTTGGATAAGATTTCCTCCGTTCCCCATGGGGAACTG

[0327] 9 TCTTGCACCACAGAGCTCCTGATCCTGGCCCATCATTGCTTCACCCTGACGTGCCACATGGAGGGC

[0328] 10 ATCATCCGAGTCCTACAGGCCGCCCACATGCTCACAGATAACCACCTGGCCCCCAGTGAGGAGTAT

[0329] 11 GGGCTGGTGGTACGGCTCCTCACTGGCATTGGAAGGTACAACGAGATGACATACATATTTGATTTG

[0330] 12 CTGCATAAAAAGCACTACTTTGAAGTGCTAATGAGGAAGAAGTTGGATCCGAGTGGTACCCTGAAA

[0331] 13 ACAGCCCTGCTGGACTACATCAAACGCTGCCGTCCTGGAGACAGTGAAAAGCACAATATGATTGCC

[0332] 14 CTGTGCTTCAGCATGTGCCGGGAGATTGGCGAGAACCACGAGGCAGCTGCCCGCATCCAACTGAAA

[0333] 15 TTGATTGAGTCTCAGCCCTGGGAGGACAGCCTCAAGGATGGGCACCAGCTGAAACAACTGCTGCTG

[0334] 16 AAGGCCCTGACTCTGATGTTGGATGCAGCAGAGAGTTATGCCAAGGACTCCTGTGTGCGACAGGCC

[0335] 17 CAGCACTGTCAGCGGCTCACCAAGTTGATAACTCTGCAGATTCACTTTCTGAACACTGGCCAGAAC

[0336] 18 ACAATGCTCATCAACTTGGGCCGCCACAAGCTGATGGACTGTATTCTGGCCCTACCTCGGTTCTAC

[0337] 19 CAGGCTTCTATTGTGGCTGAGGCCTACGATTTTGTTCCAGATTGGGCTGAAATTTTATACCAGCAA

[0338] 20 GTGATTCTTAAAGGAGACTTTAATTACTTGGAAGAATTTAAGCAGCAAAGGTTATTAAAGTCCAGT

[0339] 21 ATATTTGAAGAGATTTCCAAAAAATATAAACAACATCAGCCTACTGACATGGTCATGGAAAACCTG

[0340] 22 AAGAAATTACTCACATATTGTGAAGATGTTTACCTGTATTACAAGTTGGCATACGAACACAAGTTT

[0341] 23 TATGAAATTGTAAATGTGCTTCTGAAGGACCCTCAGACAGGTTGCTGTCTAAAGGACATGCTAGCA

[0342] 24 GGTTAG

[0343] 25

[0344] 26 SEQ ID NO : 4 - SPG11 promoter

[0345] 'Ll Aaagcgagactccatctcaaaagaaaaaaaaagtttgcttttattttgacaaataaggtcatgcat

[0346] 28 t gat caacaaatgcgggagtacctt teat gaaacatgcactgttttaggtactagccaggacagtg

[0347] 29 gtgagcaacaaagtccttgctcttttgaagtttacattcttattggacaggaactggcagaaaata

[0348] 30 tcaagtagtgataaatactatgattaaaaacaaaactaggtgagagactgcagtagaagggacctt

[0349] 31 aggttgaatggcttggggaggcctgtcaatagaggttacatttatgctgaaatgttctgaatgtgg

[0350] 32 aagatgtgttaagaaggtacagttgtcttttaggagactttaccaaaatacctattcaacccctct

[0351] 33 tctcacctcaaactctttgctcctctccttctcttgatcttggatcatacttccctgagaaaacag 1 acaggagctctctaccatctccaccccttacttaccccgtctctctgactgttacaatggatatta

[0352] 2 tttgacttcaaatttcgggtcaacacccttactatggatccacactgtaagacggtccatgctacc

[0353] 3 aaggaaatggcaccaccccagagcctgtaagcgaacagcacaataagaaaagcgaagaaaagaggc

[0354] 4 aggaactgggttccaagggagctggaggcagctggaccaagaaggcaaagagcaatgccacagagc

[0355] 5 gagaacggctgcaacggcctgctacgctaagctaggccttcaagcatgccagagcagttaagcaga

[0356] 6 gtgggactggcgtcagggcagatgtggctttgctagagaggctggtactgccccccacaggaaacg

[0357] 7 aatggaatcgaccggagacagccgagaggtctccgcgggagtcacgaggccgagcgggagccgcta

[0358] 8 cgtacaccgcgcatgcccacgacgcagtcaggttccggcgaaagtgaccggaagtAACCGCCGGGC

[0359] 9 CAAGACGCGT

[0360] 10 SEQ ID NO : 5 - Lentivirus construct (Efla promoter , SPG11

[0361] 11 sequence)

[0362] 12 CTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCC

[0363] 13 ACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGA

[0364] 14 CTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCG

[0365] 15 AACAGGGACTTGAAAGCGAAAGGGAAACCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAA

[0366] 16 GCGCGCACGGCAAGAGGCGAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGC

[0367] 17 TAGAAGGAGAGAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAA

[0368] 18 AAAATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAAGCAGG

[0369] 19 GAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTG

[0370] 20 GGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTAGCA

[0371] 21 ACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGACAAGATAGAG

[0372] 22 GAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCGGCCGGCCGCTGATCTTCAGACCTGGAG

[0373] 23 GAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGTAGTAAAAATTGAACCAT

[0374] 24 TAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAG

[0375] 25 GAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGA

[0376] 26 CGGTACAGGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG

[0377] 'Ll AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGG

[0378] 28 CTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAAACTCATTT

[0379] 29 GCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTGGAATCACA

[0380] 30 CGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATACACTCCTTAATTGAAG

[0381] 31 AAT CGCAAAACCAGC AAGAAAAGAATGAACAAGAATT AT TGGAAT T AGAT AAAT GGGC AAGT TT GT

[0382] 32 GGAATTGGTTTAACATAACAAATTGGCTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCT

[0383] 33 TGGTAGGTTTAAGAATAGTTTTTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCAC

[0384] 34 CATTATCGTTTCAGACCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAG AAGGTGGAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCGGCACTGCGTGCGCCAA

[0385] TTCTGCAGACAAATGGCAGTATTCATCCACAATTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGT

[0386] GCAGGGGAAAGAATAGTAGACATAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATT

[0387] ACAAAAATTCAAAATTTTCGGGTTTATTACAGGGACAGCAGAGATCCAGTTTGGTTAGTACCGGGC

[0388] CCGCTCTAGACGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAG

[0389] AAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAA

[0390] AGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTA

[0391] GTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGT

[0392] TCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTG

[0393] CAGTACGTGATTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCT

[0394] TAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGA

[0395] ATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGA

[0396] TGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACT

[0397] GGTATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCG

[0398] AGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGCT

[0399] CTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCA

[0400] CCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACG

[0401] CGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCC

[0402] GTCGCTTCATGTGACTCCACGGAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTT

[0403] TGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGG

[0404] GTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAG

[0405] TTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGGTG

[0406] TCGTGAGTGTCGTGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCCGCCGCGATCGCCGG

[0407] CGCGCCAGATCTCAAGCTTAACTAGCTAGCGGACCGACGCGTgccaccArGGCrGCAGAGGAAGGG

[0408] GTCGCGAGTGCTGCTTCCGCCGGCGGTAGCTGGGGCACCGCGGCCATGGGGCGGGTTCTACCGATG

[0409] CTGTTGGTGCCAGTCCCCGCCGAGGCGATGGGGCAGCTCGGCTCCCGGGCGCAGCTGCGCACACAG

[0410] CCGGAGGCTCTGGGGAGCCTGACGGCTGCGGGCAGCCTCCAAGTGCTTTCTTTGACGCCTGGCAGC

[0411] CGGGGCGGGGGTCGCTGCTGCCTGGAGGGCCCCTTCTGGCACTTTCTATGGGAGGATTCTCGTAAC

[0412] AGCAGCACACCAACTGAAAAGCCCAAACTGCTCGCTCTTGGTGAAAATTATGAACTGCTTATCTAT

[0413] GAATTTAATTTGAAAGATGGAAGATGTGATGCAACCATTTTGTATAGCTGTAGTAGGGAGGCATTG

[0414] CAAAAGCTCATTGACGATCAAGATATCAGTATTTCCTTATTGTCTTTGAGAATCCTGTCATTTCAC

[0415] AATAACACATCATTACTGTTCATCAACAAATGTGTCATCCTACATATTATATTTCCTGAAAGAGAT

[0416] GCTGCAATTAGAGTACTCAACTGTTTCACACTTCCCTTGCCTGCACAGGCAGTGGACATGATTATT

[0417] GACACGCAGCTCTGCAGAGGAATTCTTTTTGTTTTGAGTAGTTTAGGCTGGATCTACATTTTTGAT

[0418] GTTGTGGATGGTACATATGTAGCTCATGTGGATTTAGCACTTCACAAAGAAGACATGTGTAATGAG CAGCAACAGGAGCCAGCCAAGATTTCTTCATTTACTTCACTGAAAGTTTCTCAAGACCTCGATGTT

[0419] GCAGTGATTGTCAGCTCCTCCAACTCCGCAGTTGCTCTTAACTTAAATTTGTATTTCAGGCAACAC

[0420] CCAGGACACCTACTGTGTGAAAGAATACTAGAAGATCTTCCTATTCAAGGACCTAAGGGCGTAGAT

[0421] GAAGATGATCCTGTTAACTCTGCCTACAACATGAAACTGGCCAAGTTTTCCTTCCAAATTGATAGG

[0422] TCTTGGAAAGCCCAGCTATCATCATTGAATGAAACAATAAAGAACTCCAAACTGGAGGTTTCCTGT

[0423] TGTGCTCCATGGTTCCAGGATATTTTGCATTTGGAGTCACCTGAATCTGGTAACCACAGTACAAGT

[0424] GTGCAGAGCTGGGCCTTCATTCCACAGGACATAATGCATGGGCAATATAATGTTCTACAGAAAGAT

[0425] CATGCCAAGACCAGTGATCCAGGAAGATCATGGAAAATAATGCACATCAGTGAACAAGAGGAACCC

[0426] ATAGAGCTTAAATGTGTGTCTGTGACAGGATTCACTGCACTGTTTACTTGGGAAGTGGAAAGGATG

[0427] GGCTATACCATTACCCTCTGGGATTTGGAGACCCAGGGCATGCAGTGTTTTTCCCTTGGCACAAAG

[0428] TGTATTCCTGTAGACAGTAGTGGAGACCAGCAGCTGTGCTTTGTTTTGACAGAGAATGGACTCTCT

[0429] CTGATTTTGTTTGGTTTGACTCAAGAAGAGTTTTTAAACAGACTCATGATCCATGGAAGTGCCAGC

[0430] ACTGTGGACACTCTTTGTCATCTCAATGGCTGGGGAAGGTGCTCAATTCCCATACATGCACTAGAG

[0431] GCCGGGATAGAAAATCGTCAGCTGGACACAGTAAATTTCTTTTTGAAGAGCAAGGAAAATCTTTTT

[0432] AATCCATCCTCAAAATCTTCTGTATCTGATCAGTTTGATCACTTGTCATCCCATTTATATTTAAGA

[0433] AATGTGGAAGAGCTGATACCAGCATTGGATTTACTTTGCTCGGCAATTAGAGAAAGTTATTCTGAA

[0434] CCCCAAAGCAAACACTTTTCAGAACAATTGCTTAATCTTACACTGTCTTTCCTTAACAACCAAATA

[0435] AAGGAGCTTTTCATTCACACTGAAGAACTAGATGAACATCTGCAAAAAGGAGTGAACATTTTGACT

[0436] AGCTACATTAATGAACTTCGAACCTTCATGATAAAGTTTCCTTGGAAGCTAACAGATGCTATAGAT

[0437] GAATATGATGTACATGAAAATGTCCCCAAAGTAAAGGAGAGCAATATATGGAAGAAACTCAGCTTT

[0438] GAGGAAGTTATTGCCAGCGCCATTTTAAACAACAAAATACCAGAGGCACAGACTTTCTTCAGGATT

[0439] GATAGTCATTCTGCTCAAAAACTTGAGGAGCTTATTGGCATAGGCCTAAATTTGGTCTTTGACAAT

[0440] TTAAAAAAGAACAATATAAAGGAAGCCTCTGAACTTTTGAAGAATATGGGGTTTGATGTAAAAGGC

[0441] CAATTGCTCAAGATCTGCTTCTATACAACTAATAAAAATATACGTGACTTTTTGGTTGAAATTTTA

[0442] AAAGAAAAAAATTATTTTTCTGAAAAAGAGAAAAGAACTATAGACTTCGTGCATCAAGTTGAGAAG

[0443] CTTTATTTGGGACATTTCCAAGAAAATATGCAAATCCAGTCATTTCCCAGGTACTGGATAAAGGAA

[0444] CAAGATTTTTTCAAGCACAAGTCTGTTTTGGACTCATTCCTGAAATATGATTGTAAAGATGAATTT

[0445] AACAAACAGGACCATAGAATTGTGTTAAATTGGGCTCTGTGGTGGGATCAACTAACACAAGAATCC

[0446] ATCCTTCTCCCCAGGATAAGTCCAGAAGAATACAAATCATATTCCCCTGAAGCCCTCTGGAGATAC

[0447] CTCACAGCTCGCCATGATTGGTTAAACATTATCTTATGGATTGGAGAATTTCAAACCCAGCATAGT

[0448] TATGCTTCACTTCAGCAGAACAAATGGCCCCTTCTGACTGTTGATGTTATTAACCAGAATACTTCC

[0449] TGTAACAACTACATGAGGAATGAAATTTTAGATAAGCTGGCCAGGAATGGGGTTTTTTTGGCATCT

[0450] GAACTGGAAGACTTTGAATGCTTCCTCCTAAGACTGAGCCGTATTGGAGGTGTAATACAGGATACC

[0451] CTCCCTGTTCAAAACTACAAGACCAAAGAAGGTTGGGATTTCCATTCTCAATTCATTCTCTATTGT

[0452] TTGGAGCACAGTCTGCAGCATCTTCTTTATGTCTACCTTGACTGTTACAAACTTAGTCCTGAAAAT TGTCCCTTTTTGGAAAAAAAAGAGTTACATGAAGCACACCCTTGGTTTGAATTTTTAGTTCAGTGT CGACAAGTTGCCAGTAACTTAACAGATCCCAAACTGATCTTCCAGGCTAGCCTTGCAAATGCTCAG ATTTTGATTCCCACCAATCAGGCCAGTGTAAGCAGTATGCTATTGGAAGGACATACCCTCCTGGCC CTTGCTACTACAATGTATTCTCCTGGGGGTGTCAGTCAGGTTGTTCAGAATGAAGAAAATGAAAAC TGTTTGAAGAAAGTGGATCCCCAGCTATTGAAGATGGCATTAACTCCTTACCCCAAGCTAAAAACT GCTCTCTTCCCACAGTGCACTCCTCCTAGTGTCCTGCCATCTGATATTACAATCTACCACCTTATT CAGTCATTATCACCCTTTGATCCTAGCAGATTGTTTGGCTGGCAGTCTGCTAACACACTAGCTATA GGAGATGCATGGAGTCATCTCCCACATTTCTCTAGCCCTGACCTGGTTAATAAATATGCTATAGTG GAACGTCTGAATTTTGCTTATTATTTACATAATGGGCGGCCATCATTTGCATTTGGTACTTTTCTG GTCCAGGAATTAATCAAGAGCAAGACTCCCAAGCAGCTGATCCAGCAAGTAGGCAATGAAGCCTAT GTTATAGGGCTCTCCTCCTTCCACATACCTTCAATAGGAGCTGCATGTGTTTGTTTCTTAGAATTG CTTGGCCTTGACAGCCTCAAGCTCAGAGTTGATATGAAAGTGGCCAATATAATTTTGAGCTACAAG TGC AG AAA TGAAGATGC TCAG TACAGCTTTA TCAGAGAGTC TGTAGCCGAAAAA CT A TCTAAAC TA GCTGATGGTGAAAAGACAACCACAGAAGAATTGCTTGTTCTCTTAGAAGAAGGTACATGGAACAGC ATTCAGCAACAGGAAATAAAGAGGTTATCCAGTGAATCTAGCAGCCAATGGGCATTAGTGGTGCAG TTCTGCAGGCTACACAATATGAAACTAAGCATATCTTACCTTAGAGAATGTGCCAAAGCAAATGAT TGGCTGCAGTTCATTATTCACAGCCAACTCCACAACTACCACCCAGCAGAGGTGAAATCCCTTATC CAGTACTTCAGCCCAGTCATTCAAGACCACTTAAGGCTGGCTTTTGAGAACTTGCCCTCAGTGCCC ACCTCCAAAATGGACAGCGATCAAGTCTGCAATAAGTGCCCCCAGGAACTTCAAGGAAGCAAACAA GAGATGACCGATTTATTTGAAATTCTGCTCCAATGCTCAGAGGAGCCAGACTCCTGGCACTGGCTT CTGGTTGAAGCAGTGAAACAACAGGCCCCTATCCTCAGTGTTCTGGCCTCATGTCTCCAGGGTGCC AGTGCCATTTCTTGTCTCTGTGTTTGGATCATCACTTCTGTGGAGGACAATGTTGCAACTGAAGCA ATGGGACACATTCAGGACTCAACAGAGGACCATACCTGGAACCTTGAGGATCTTTCAGTCATCTGG AGAACATTATTAACAAGACAAAAGAGCAAAACTCTCATCAGAGGTTTCCAGCTTTTCTTTAAGGAT TCCCCGTTACTACTGGTGATGGAGATGTATGAACTGTGTATGTTCTTCAGGAATTATAAAGAAGCT GAAGCTAAACTTCTGGAGTTTCAGAAGAGCCTTGAAACGCTTAACACAGCAGCCACAAAGGTCCAC CCTGTCATCCCTGCCATGTGGCTGGAGGATCAGGTGTGTTTCCTTTTGAAGCTTATGCTACAGCAG TGTAAGACCCAGTATGAGCTGGGGAAGCTTTTACAGCTCTTTGTTGAAAGAGAGCATCTCTTCTCT GATGGTCCAGATGTGAAAAAGCTTTGCATCCTTTGCCAGATTTTGAAGGATACATCCATAGCCATT AATCATACAATTATTACCAGCTACAGCATTGAGAATCTTCAGCATGAATGTAGATCTATTTTGGAA AGACTGCAGACAGATGGACAATTCGCTTTGGCCAGGAGGGTAGCAGAATTAGCTGAGTTACCTGTG GACAACTTGGTTATTAAAGAGATAACACAGGAAATGCAGACCCTAAAACACATTGAACAGTGGTCA CTAAAACAAGCAAGAATTGACTTCTGGAAAAAATGCCATGAGAATTTTAAGAAAAATTCAATTTCA AGCAAAGCAGCTTCTTCCTTTTTCTCAACCCAGGCCCATGTGGCATGTGAGCACCCAACTGGATGG AGCAGCATGGAGGAGCGCCATCTGCTGCTCACCTTGGCAGGGCACTGGCTTGCCCAGGAGGACGTG GTGCCCTTGGATAAGCTGGAGGAGCTGGAGAAGCAGATCTGGCTGTGCCGCATCACCCAGCACACT CTTGGAAGAAATCAGGAGGAAACAGAGCCCAGATTTTCTCGACAGATCTCAACTAGTGGTGAACTT TCCTTTGATAGTTTAGCCAGTGAGTTTTCCTTCTCCAAGTTGGCTGCTCTGAACACATCAAAATAC TTAGAACTTAACAGCCTTCCA TCCAAAGAGACA TGCGAGAA TAG A TTGGA TTGGAAAGAGCAGGAG TCACTAAACTTTTTGATTGGGCGCCTACTGGATGATGGCTGTGTGCATGAAGCAAGTAGAGTATGC CGGTATTTTCATTTTTATAATCCAGATGTCGCCTTGGTATTGCACTGCAGAGCACTGGCCTCAGGG GAAGCTAGTATGGAGGATCTGCACCCAGAGATCCATGCTCTCCTACAAAGTGCTGAGCTGCTTGAG GAAGAAGCACCCGACATTCCCCTAAGGAGAGTCCACAGCACTTCAAGTCTGGATAGTCAGAAGTTT GTGACAGTGCCCTCCAGTAATGAAGTGGTAACTAACCTGGAAGTGCTGACAAGCAAATGCCTCCAT GGGAAGAACTACTGTCGACAGGTCCTCTGTCTGTATGATCTTGCCAAGGAGTTGGGCTGTTCCTAC ACAGATGTTGCTGCTCAGGATGGTGAAGCCATGCTCCGGAAAATCTTGGCCTCTCAGCAGCCTGAC CGATGCAAACGAGCCCAGGCCTTCATCAGCACACAGGGCCTTAAGCCAGATACTGTGGCTGAACTC GTGGCAGAAGAGGTGACACGGGAGCTGCTTACTTCATCACAGGGAACAGGACATAAGCAGATGTTC AACCCAACAGAGGAAAGCCAGACATTTCTTCAGCTGACCACTCTGTGTCAAGACCGCACATTGGTA GGCATGAAGTTGTTGGATAAGATTTCCTCCGTTCCCCATGGGGAACTGTCTTGCACCACAGAGCTC CTGATCCTGGCCCATCATTGCTTCACCCTGACGTGCCACATGGAGGGCATCATCCGAGTCCTACAG GCCGCCCACATGCTCACAGATAACCACCTGGCCCCCAGTGAGGAGTATGGGCTGGTGGTACGGCTC CTCACTGGCATTGGAAGGTACAACGAGATGACATACATATTTGATTTGCTGCATAAAAAGCACTAC TTTGAAGTGCTAATGAGGAAGAAGTTGGATCCGAGTGGTACCCTGAAAACAGCCCTGCTGGACTAC ATCAAACGCTGCCGTCCTGGAGACAGTGAAAAGCACAATATGATTGCCCTGTGCTTCAGCATGTGC CGGGAGA TTGGCGAGAACCACGAGGCAGCTGCCCGCA TCCAACTGAAA TTGA TTGAGTCTCAGCCC TGGGAGGACAGCCTCAAGGATGGGCACCAGCTGAAACAACTGCTGCTGAAGGCCCTGACTCTGATG TTGGATGCAGCAGAGAGTTATGCCAAGGACTCCTGTGTGCGACAGGCCCAGCACTGTCAGCGGCTC ACCAAGTTGATAACTCTGCAGATTCACTTTCTGAACACTGGCCAGAACACAATGCTCATCAACTTG GGCCGCCACAAGCTGATGGACTGTATTCTGGCCCTACCTCGGTTCTACCAGGCTTCTATTGTGGCT GAGGCCTACGATTTTGTTCCAGATTGGGCTGAAATTTTATACCAGCAAGTGATTCTTAAAGGAGAC TTTAATTACTTGGAAGAATTTAAGCAGCAAAGGTTATTAAAGTCCAGTATATTTGAAGAGATTTCC AAAAAATATAAACAACATCAGCCTACTGACATGGTCATGGAAAACCTGAAGAAATTACTCACATAT TGTGAAGATGTTTACCTGTATTACAAGTTGGCATACGAACACAAGTTTTATGAAATTGTAAATGTG CTTCTGAAGGACCCTCAGACAGGTTGCTGTCTAAAGGACATGCTAGCAGGTTAGGCGGCCTAAGGA TGACGACGATAAATTCGTCGAGCACCACCACCACCACCACTAATAAGGTTTATCCGATCCACCGGA TCTAGATAAGATAAACGGCCGGCCGCGGTCTGTACAAGTAGGATTCGTCGAGGGACCTAATAACTT CGTATAGCATACATTATACGAAGTTATACATGTTTAAGGGTTCCGGTTCCACTAGGTACAATTCGA TATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAA CTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTC 1 CCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTG

[0453] 2 GCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGG

[0454] 3 CATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGA

[0455] 4 ACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGT

[0456] 5 GGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCG

[0457] 6 CGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCT

[0458] 7 GCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGC

[0459] 8 CGCCTCCCCGCATCGATACCGTCGACCTCGATCGAGACCTAGAAAAACATGGAGCAATCACAAGTA

[0460] 9 GCAATACAGCAGCTACCAATGCTGATTGTGCCTGGCTAGAAGCACAAGAGGAGGAGGAGGTGGGTT

[0461] 10 TTCCAGTCACACCTCAGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCACT

[0462] 11 TTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATATCCTTGATC

[0463] 12 TGTGGATTCCGGACTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCT

[0464] 13 AACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCC

[0465] 14 GTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTA

[0466] 15 GCAGCATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTT

[0467] 16 SEQ ID NO : 6 - Lentivirus construct (UBC promoter , SPG11 sequence)

[0468] 17 CTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCC

[0469] 18 ACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGA

[0470] 19 CTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCG

[0471] 20 AACAGGGACTTGAAAGCGAAAGGGAAACCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAA

[0472] 21 GCGCGCACGGCAAGAGGCGAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGC

[0473] 22 TAGAAGGAGAGAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAA

[0474] 23 AAAATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAAGCAGG

[0475] 24 GAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTG

[0476] 25 GGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTAGCA

[0477] 26 ACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGACAAGATAGAG

[0478] 'Ll GAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCGGCCGGCCGCTGATCTTCAGACCTGGAG

[0479] 28 GAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGTAGTAAAAATTGAACCAT

[0480] 29 TAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAG

[0481] 30 GAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGA

[0482] 31 CGGTACAGGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG

[0483] 32 AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGG

[0484] 33 CTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAAACTCATTT

[0485] 34 GCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTGGAATCACA CGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATACACTCCTTAATTGAAG

[0486] AAT CGCAAAACCAGC AAGAAAAGAATGAACAAGAATT AT TGGAAT T AGAT AAAT GGGC AAGT TT GT

[0487] GGAATTGGTTTAACATAACAAATTGGCTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCT

[0488] TGGTAGGTTTAAGAATAGTTTTTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCAC

[0489] CATTATCGTTTCAGACCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAG

[0490] AAGGTGGAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCGGCACTGCGTGCGCCAA

[0491] TTCTGCAGACAAATGGCAGTATTCATCCACAATTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGT

[0492] GCAGGGGAAAGAATAGTAGACATAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATT

[0493] ACAAAAATTCAAAATTTTCGGGTTTATTACAGGGACAGCAGAGATCCAGTTTGGTTAGTACCGGGC

[0494] CCGCTCTAGACCGCGGCCTCCGCGCCGGGTTTTGGCGCCTCCCGCGGGCGCCCCCCTCCTCACGGC

[0495] GAGCGCTGCCACGTCAGACGAAGGGCGCAGCGAGCGTCCTGATCCTTCCGCCCGGACGCTCAGGAC

[0496] AGCGGCCCGCTGCTCATAAGACTCGGCCTTAGAACCCCAGTATCAGCAGAAGGACATTTTAGGACG

[0497] GGACTTGGGTGACTCTAGGGCACTGGTTTTCTTTCCAGAGAGCGGAACAGGCGAGGAAAAGTAGTC

[0498] CCTTCTCGGCGATTCTGCGGAGGGATCTCCGTGGGGCGGTGAACGCCGATGATTATATAAGGACGC

[0499] GCCGGGTGTGGCACAGCTAGTTCCGTCGCAGCCGGGATTTGGGTCGCAGTTCTTGTTTGTGGATTG

[0500] CTGTGATCGTCACTTGGTGAGTAGCGGGCTGCTGGGCTGGCCGGGGCTTTCGTGGCCGCCGGGCCG

[0501] CTCGGTGGGACGGAAGCGTGTGGAGAGACCGCCAAGGGCTGTAGTCTGGGTCCGCGAGCAAGGTTG

[0502] CCCTGAACTGGGGGTTGGGGGGAGCGCAGCAAAATGGCGGCTGTTCCCGAGTCTTGAATAGAACCT

[0503] TCGCTAATGCGGGAAAGCTCTTATTCGGGTGAGATGGGCTGGGGCACCATCTGGGGACCCTGACGT

[0504] GAAGTTTGTCACTGACTGGAGAACTCGGTTTGTCGTCTGTTGCGGGGGCGGCAGTTATGGCGGTGC

[0505] CGTTGGGCAGTGCACCCGTACCTTTGGGAGCGCGCGCCCTCGTCGTGTCGTGACGTCACCCGTTCT

[0506] GTTGGCTTATAATGCAGGGTGGGGCCACCTGCCGGTAGGTGTGCGGTAGGCTTTTCTCCGTCGCAG

[0507] GACGCAGGGTTCGGGCCTAGGGTAGGCTCTCCTGAATCGACAGGCGCCGGACCTCTGGTGAGGGGA

[0508] GGGATAAGTGAGGCGTCAGTTTCTTTGGTCGGTTTTATGTACCTATCTTCTTAAGTAGCTGAAGCT

[0509] CCGGTTTTGAACTATGCGCTCGGGGTTGGCGAGTGTGTTTTGTGAAGTTTTTTAGGCACCTTTTGA

[0510] AATGTAATCATTTGGGTCAATATGTAATTTTCAGTGTTAGACTAGTAAATTGTCCGCTAAATTCTG

[0511] GCCGTTTTTGGCTTTTTTGTTAGACGGTACCGAGGAGATCTGCCGCCGCGATCGCCGGCGCGCCAG

[0512] ATCTCACGCTTAACTAGCTAGCGGACCGACGCGTgccaccATGGCTGCAGAGGAAGGGGTCGCGAG

[0513] TGCTGCTTCCGCCGGCGGTAGCTGGGGCACCGCGGCCATGGGGCGGGTTCTACCGATGCTGTTGGT

[0514] GCCAGTCCCCGCCGAGGCGATGGGGCAGCTCGGCTCCCGGGCGCAGCTGCGCACACAGCCGGAGGC

[0515] TCTGGGGAGCCTGACGGCTGCGGGCAGCCTCCAAGTGCTTTCTTTGACGCCTGGCAGCCGGGGCGG

[0516] GGGTCGCTGCTGCCTGGAGGGCCCCTTCTGGCACTTTCTATGGGAGGATTCTCGTAACAGCAGCAC

[0517] ACCAACTGAAAAGCCCAAACTGCTCGCTCTTGGTGAAAATTATGAACTGCTTATCTATGAATTTAA

[0518] TTTGAAAGATGGAAGATGTGATGCAACCATTTTGTATAGCTGTAGTAGGGAGGCATTGCAAAAGCT

[0519] CATTGACGATCAAGATATCAGTATTTCCTTATTGTCTTTGAGAATCCTGTCATTTCACAATAACAC ATCATTACTGTTCATCAACAAATGTGTCATCCTACATATTATATTTCCTGAAAGAGATGCTGCAAT

[0520] TAGAGTACTCAACTGTTTCACACTTCCCTTGCCTGCACAGGCAGTGGACATGATTATTGACACGCA

[0521] GCTCTGCAGAGGAATTCTTTTTGTTTTGAGTAGTTTAGGCTGGATCTACATTTTTGATGTTGTGGA

[0522] TGGTACATATGTAGCTCATGTGGATTTAGCACTTCACAAAGAAGACATGTGTAATGAGCAGCAACA

[0523] GGAGCCAGCCAAGATTTCTTCATTTACTTCACTGAAAGTTTCTCAAGACCTCGATGTTGCAGTGAT

[0524] TGTCAGCTCCTCCAACTCCGCAGTTGCTCTTAACTTAAATTTGTATTTCAGGCAACACCCAGGACA

[0525] CCTACTGTGTGAAAGAATACTAGAAGATCTTCCTATTCAAGGACCTAAGGGCGTAGATGAAGATGA

[0526] TCCTGTTAACTCTGCCTACAACATGAAACTGGCCAAGTTTTCCTTCCAAATTGATAGGTCTTGGAA

[0527] AGCCCAGCTATCATCATTGAATGAAACAATAAAGAACTCCAAACTGGAGGTTTCCTGTTGTGCTCC

[0528] ATGGTTCCAGGATATTTTGCATTTGGAGTCACCTGAATCTGGTAACCACAGTACAAGTGTGCAGAG

[0529] CTGGGCCTTCATTCCACAGGACATAATGCATGGGCAATATAATGTTCTACAGAAAGATCATGCCAA

[0530] GACCAGTGATCCAGGAAGATCATGGAAAATAATGCACATCAGTGAACAAGAGGAACCCATAGAGCT

[0531] TAAATGTGTGTCTGTGACAGGATTCACTGCACTGTTTACTTGGGAAGTGGAAAGGATGGGCTATAC

[0532] CATTACCCTCTGGGATTTGGAGACCCAGGGCATGCAGTGTTTTTCCCTTGGCACAAAGTGTATTCC

[0533] TGTAGACAGTAGTGGAGACCAGCAGCTGTGCTTTGTTTTGACAGAGAATGGACTCTCTCTGATTTT

[0534] GTTTGGTTTGACTCAAGAAGAGTTTTTAAACAGACTCATGATCCATGGAAGTGCCAGCACTGTGGA

[0535] CACTCTTTGTCATCTCAATGGCTGGGGAAGGTGCTCAATTCCCATACATGCACTAGAGGCCGGGAT

[0536] AGAAAATCGTCAGCTGGACACAGTAAATTTCTTTTTGAAGAGCAAGGAAAATCTTTTTAATCCATC

[0537] CTCAAAATCTTCTGTATCTGATCAGTTTGATCACTTGTCATCCCATTTATATTTAAGAAATGTGGA

[0538] AGAGCTGATACCAGCATTGGATTTACTTTGCTCGGCAATTAGAGAAAGTTATTCTGAACCCCAAAG

[0539] CAAACACTTTTCAGAACAATTGCTTAATCTTACACTGTCTTTCCTTAACAACCAAATAAAGGAGCT

[0540] TTTCATTCACACTGAAGAACTAGATGAACATCTGCAAAAAGGAGTGAACATTTTGACTAGCTACAT

[0541] TAATGAACTTCGAACCTTCATGATAAAGTTTCCTTGGAAGCTAACAGATGCTATAGATGAATATGA

[0542] TGTACATGAAAATGTCCCCAAAGTAAAGGAGAGCAATATATGGAAGAAACTCAGCTTTGAGGAAGT

[0543] TATTGCCAGCGCCATTTTAAACAACAAAATACCAGAGGCACAGACTTTCTTCAGGATTGATAGTCA

[0544] TTCTGCTCAAAAACTTGAGGAGCTTATTGGCATAGGCCTAAATTTGGTCTTTGACAATTTAAAAAA

[0545] GAACAATATAAAGGAAGCCTCTGAACTTTTGAAGAATATGGGGTTTGATGTAAAAGGCCAATTGCT

[0546] CAAGATCTGCTTCTATACAACTAATAAAAATATACGTGACTTTTTGGTTGAAATTTTAAAAGAAAA

[0547] AAATTATTTTTCTGAAAAAGAGAAAAGAACTATAGACTTCGTGCATCAAGTTGAGAAGCTTTATTT

[0548] GGGACATTTCCAAGAAAATATGCAAATCCAGTCATTTCCCAGGTACTGGATAAAGGAACAAGATTT

[0549] TTTCAAGCACAAGTCTGTTTTGGACTCATTCCTGAAATATGATTGTAAAGATGAATTTAACAAACA

[0550] GGACCATAGAATTGTGTTAAATTGGGCTCTGTGGTGGGATCAACTAACACAAGAATCCATCCTTCT

[0551] CCCCAGGATAAGTCCAGAAGAATACAAATCATATTCCCCTGAAGCCCTCTGGAGATACCTCACAGC

[0552] TCGCCATGATTGGTTAAACATTATCTTATGGATTGGAGAATTTCAAACCCAGCATAGTTATGCTTC

[0553] ACTTCAGCAGAACAAATGGCCCCTTCTGACTGTTGATGTTATTAACCAGAATACTTCCTGTAACAA CTACATGAGGAATGAAATTTTAGATAAGCTGGCCAGGAATGGGGTTTTTTTGGCATCTGAACTGGA

[0554] AGACTTTGAATGCTTCCTCCTAAGACTGAGCCGTATTGGAGGTGTAATACAGGATACCCTCCCTGT

[0555] TCAAAACTACAAGACCAAAGAAGGTTGGGATTTCCATTCTCAATTCATTCTCTATTGTTTGGAGCA

[0556] CAGTCTGCAGCATCTTCTTTATGTCTACCTTGACTGTTACAAACTTAGTCCTGAAAATTGTCCCTT

[0557] TTTGGAAAAAAAAGAGTTACATGAAGCACACCCTTGGTTTGAATTTTTAGTTCAGTGTCGACAAGT

[0558] TGCCAGTAACTTAACAGATCCCAAACTGATCTTCCAGGCTAGCCTTGCAAATGCTCAGATTTTGAT

[0559] TCCCACCAATCAGGCCAGTGTAAGCAGTATGCTATTGGAAGGACATACCCTCCTGGCCCTTGCTAC

[0560] TACAATGTATTCTCCTGGGGGTGTCAGTCAGGTTGTTCAGAATGAAGAAAATGAAAACTGTTTGAA

[0561] GAAAGTGGATCCCCAGCTATTGAAGATGGCATTAACTCCTTACCCCAAGCTAAAAACTGCTCTCTT

[0562] CCCACAGTGCACTCCTCCTAGTGTCCTGCCATCTGATATTACAATCTACCACCTTATTCAGTCATT

[0563] ATCACCCTTTGATCCTAGCAGATTGTTTGGCTGGCAGTCTGCTAACACACTAGCTATAGGAGATGC

[0564] ATGGAGTCATCTCCCACATTTCTCTAGCCCTGACCTGGTTAATAAATATGCTATAGTGGAACGTCT

[0565] GAATTTTGCTTATTATTTACATAATGGGCGGCCATCATTTGCATTTGGTACTTTTCTGGTCCAGGA

[0566] ATTAATCAAGAGCAAGACTCCCAAGCAGCTGATCCAGCAAGTAGGCAATGAAGCCTATGTTATAGG

[0567] GCTCTCCTCCTTCCACATACCTTCAATAGGAGCTGCATGTGTTTGTTTCTTAGAATTGCTTGGCCT

[0568] TGACAGCCTCAAGCTCAGAGTTGATATGAAAGTGGCCAATATAATTTTGAGCTACAAGTGCAGAAA

[0569] TGAAGATGCTCAGTACAGCTTTATCAGAGAGTCTGTAGCCGAAAAACTATCTAAACTAGCTGATGG

[0570] TGAAAAGACAACCACAGAAGAATTGCTTGTTCTCTTAGAAGAAGGTACATGGAACAGCATTCAGCA

[0571] ACAGGAAATAAAGAGGTTATCCAGTGAATCTAGCAGCCAATGGGCATTAGTGGTGCAGTTCTGCAG

[0572] GCTACACAATATGAAACTAAGCATATCTTACCTTAGAGAATGTGCCAAAGCAAATGATTGGCTGCA

[0573] GTTCATTATTCACAGCCAACTCCACAACTACCACCCAGCAGAGGTGAAATCCCTTATCCAGTACTT

[0574] CAGCCCAGTCATTCAAGACCACTTAAGGCTGGCTTTTGAGAACTTGCCCTCAGTGCCCACCTCCAA

[0575] AATGGACAGCGATCAAGTCTGCAATAAGTGCCCCCAGGAACTTCAAGGAAGCAAACAAGAGATGAC

[0576] CGATTTATTTGAAATTCTGCTCCAATGCTCAGAGGAGCCAGACTCCTGGCACTGGCTTCTGGTTGA

[0577] AGCAGTGAAACAACAGGCCCCTATCCTCAGTGTTCTGGCCTCATGTCTCCAGGGTGCCAGTGCCAT

[0578] TTCTTGTCTCTGTGTTTGGATCATCACTTCTGTGGAGGACAATGTTGCAACTGAAGCAATGGGACA

[0579] CATTCAGGACTCAACAGAGGACCATACCTGGAACCTTGAGGATCTTTCAGTCATCTGGAGAACATT

[0580] ATTAACAAGACAAAAGAGCAAAACTCTCATCAGAGGTTTCCAGCTTTTCTTTAAGGATTCCCCGTT

[0581] ACTACTGGTGATGGAGATGTATGAACTGTGTATGTTCTTCAGGAATTATAAAGAAGCTGAAGCTAA

[0582] ACTTCTGGAGTTTCAGAAGAGCCTTGAAACGCTTAACACAGCAGCCACAAAGGTCCACCCTGTCAT

[0583] CCCTGCCATGTGGCTGGAGGATCAGGTGTGTTTCCTTTTGAAGCTTATGCTACAGCAGTGTAAGAC

[0584] CCAGTATGAGCTGGGGAAGCTTTTACAGCTCTTTGTTGAAAGAGAGCATCTCTTCTCTGATGGTCC

[0585] AGATGTGAAAAAGCTTTGCATCCTTTGCCAGATTTTGAAGGATACATCCATAGCCATTAATCATAC

[0586] AATTATTACCAGCTACAGCATTGAGAATCTTCAGCATGAATGTAGATCTATTTTGGAAAGACTGCA

[0587] GACAGATGGACAATTCGCTTTGGCCAGGAGGGTAGCAGAATTAGCTGAGTTACCTGTGGACAACTT GGTTATTAAAGAGATAACACAGGAAATGCAGACCCTAAAACACATTGAACAGTGGTCACTAAAACA

[0588] AGCAAGAATTGACTTCTGGAAAAAATGCCATGAGAATTTTAAGAAAAATTCAATTTCAAGCAAAGC

[0589] AGCTTCTTCCTTTTTCTCAACCCAGGCCCATGTGGCATGTGAGCACCCAACTGGATGGAGCAGCAT

[0590] GGAGGAGCGCCATCTGCTGCTCACCTTGGCAGGGCACTGGCTTGCCCAGGAGGACGTGGTGCCCTT

[0591] GGATAAGCTGGAGGAGCTGGAGAAGCAGATCTGGCTGTGCCGCATCACCCAGCACACTCTTGGAAG

[0592] AAATCAGGAGGAAACAGAGCCCAGATTTTCTCGACAGATCTCAACTAGTGGTGAACTTTCCTTTGA

[0593] TAGTTTAGCCAGTGAGTTTTCCTTCTCCAAGTTGGCTGCTCTGAACACATCAAAATACTTAGAACT

[0594] TAACAGCCTTCCATCCAAAGAGACATGCGAGAATAGATTGGATTGGAAAGAGCAGGAGTCACTAAA

[0595] CTTTTTGATTGGGCGCCTACTGGATGATGGCTGTGTGCATGAAGCAAGTAGAGTATGCCGGTATTT

[0596] TCATTTTTATAATCCAGATGTCGCCTTGGTATTGCACTGCAGAGCACTGGCCTCAGGGGAAGCTAG

[0597] TATGGAGGATCTGCACCCAGAGATCCATGCTCTCCTACAAAGTGCTGAGCTGCTTGAGGAAGAAGC

[0598] ACCCGACATTCCCCTAAGGAGAGTCCACAGCACTTCAAGTCTGGATAGTCAGAAGTTTGTGACAGT

[0599] GCCCTCCAGTAATGAAGTGGTAACTAACCTGGAAGTGCTGACAAGCAAATGCCTCCATGGGAAGAA

[0600] CTACTGTCGACAGGTCCTCTGTCTGTATGATCTTGCCAAGGAGTTGGGCTGTTCCTACACAGATGT

[0601] TGCTGCTCAGGATGGTGAAGCCATGCTCCGGAAAATCTTGGCCTCTCAGCAGCCTGACCGATGCAA

[0602] ACGAGCCCAGGCCTTCATCAGCACACAGGGCCTTAAGCCAGATACTGTGGCTGAACTCGTGGCAGA

[0603] AGAGGTGACACGGGAGCTGCTTACTTCATCACAGGGAACAGGACATAAGCAGATGTTCAACCCAAC

[0604] AGAGGAAAGCCAGACATTTCTTCAGCTGACCACTCTGTGTCAAGACCGCACATTGGTAGGCATGAA

[0605] GTTGTTGGATAAGATTTCCTCCGTTCCCCATGGGGAACTGTCTTGCACCACAGAGCTCCTGATCCT

[0606] GGCCCATCATTGCTTCACCCTGACGTGCCACATGGAGGGCATCATCCGAGTCCTACAGGCCGCCCA

[0607] CATGCTCACAGATAACCACCTGGCCCCCAGTGAGGAGTATGGGCTGGTGGTACGGCTCCTCACTGG

[0608] CATTGGAAGGTACAACGAGATGACATACATATTTGATTTGCTGCATAAAAAGCACTACTTTGAAGT

[0609] GCTAATGAGGAAGAAGTTGGATCCGAGTGGTACCCTGAAAACAGCCCTGCTGGACTACATCAAACG

[0610] CTGCCGTCCTGGAGACAGTGAAAAGCACAATATGATTGCCCTGTGCTTCAGCATGTGCCGGGAGAT

[0611] TGGCGAGAACCACGAGGCAGCTGCCCGCATCCAACTGAAATTGATTGAGTCTCAGCCCTGGGAGGA

[0612] CAGCCTCAAGGATGGGCACCAGCTGAAACAACTGCTGCTGAAGGCCCTGACTCTGATGTTGGATGC

[0613] AGCAGAGAGTTATGCCAAGGACTCCTGTGTGCGACAGGCCCAGCACTGTCAGCGGCTCACCAAGTT

[0614] GATAACTCTGCAGATTCACTTTCTGAACACTGGCCAGAACACAATGCTCATCAACTTGGGCCGCCA

[0615] CAAGCTGATGGACTGTATTCTGGCCCTACCTCGGTTCTACCAGGCTTCTATTGTGGCTGAGGCCTA

[0616] CGATTTTGTTCCAGATTGGGCTGAAATTTTATACCAGCAAGTGATTCTTAAAGGAGACTTTAATTA

[0617] CTTGGAAGAATTTAAGCAGCAAAGGTTATTAAAGTCCAGTATATTTGAAGAGATTTCCAAAAAATA

[0618] TAAACAACATCAGCCTACTGACATGGTCATGGAAAACCTGAAGAAATTACTCACATATTGTGAAGA

[0619] TGTTTACCTGTATTACAAGTTGGCATACGAACACAAGTTTTATGAAATTGTAAATGTGCTTCTGAA

[0620] GGACCCTCAGACAGGTTGCTGTCTAAAGGACATGCTAGCAGGTTAGGCGGCCGAAGGATGACGACG

[0621] ATAAATTCGTCGAGCACCACCACCACCACCACTAATAAGGTTTATCCGATCCACCGGATCTAGATA AGATAAACGGCCGGCCGCGGTCTGTACAAGTAGGATTCGTCGAGGGACCTAATAACTTCGTATAGC

[0622] ATACATTATACGAAGTTATACATGTTTAAGGGTTCCGGTTCCACTAGGTACAATTCGATATCAAGC

[0623] TTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTG

[0624] CTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGG

[0625] CTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTG

[0626] TCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCA

[0627] CCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCG

[0628] CCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGT

[0629] CGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGT

[0630] CCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTC

[0631] TGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCC

[0632] CGCATCGATACCGTCGACCTCGATCGAGACCTAGAAAAACATGGAGCAATCACAAGTAGCAATACA

[0633] GCAGCTACCAATGCTGATTGTGCCTGGCTAGAAGCACAAGAGGAGGAGGAGGTGGGTTTTCCAGTC

[0634] ACACCTCAGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCACTTTTTAAAA

[0635] GAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATATCCTTGATCTGTGGATT

[0636] CCGGACTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGG

[0637] AACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTG

[0638] TGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAG

Claims

CLAIMS:

1. A lentivirus vector comprising an SPG11 gene under the control of a promoter.

2. The lentivirus vector of claim 1, wherein the SPG11 gene encodes a polypeptide having the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 1.

3. The lentivirus vector of claim 1 or 2, wherein SPG11 gene is a cDNA sequence.

4. The lentivirus vector of claim 3, wherein the cDNA sequence comprises the nucleotide sequence of SEQ ID NO: 2.

5. The lentivirus vector of claim 2, wherein the SPG11 gene comprises one or more introns.

6. The lentivirus vector of any one of claims 1 to 5, wherein the nucleotide sequence of the SPG11 gene is codon optimized.

7. The lentivirus vector of any one of claims 1 to 6, wherein the SPG11 gene comprises a 5'- and / or 3'- untranslated region.

8. The lentivirus vector of claim 7, wherein the 5' and / or 3' untranslated regions comprise naturally-occurring sequences from the SPG11 gene.

9. The lentivirus vector of claim 7, wherein the 5' and / or 3' untranslated regions comprise sequences that are not from the wild-type SPG11 gene.

10. The lentivirus vector of any one of claims 7 to 9, wherein the SPG11 gene comprises a poly adenylation signal.

11. The lentivirus vector of claim 10, wherein the polyadenylation signal is derived from the bovine growth hormone (BGH) gene or simian virus 40 (SV40).

12. The lentivirus vector of any one of claims 1 to 6, wherein the SPG11 gene does not include substantial untranslated regions, and optionally consists essentially of or consists of the SPG11 coding sequence and optionally a polyadenylation signal.

13. The lentivirus vector of any one of claims 1 to 12, wherein the promoter is a heterologous promoter.

14. The lentivirus vector of claim 13, wherein the promoter is a ubiquitous promoter.

15. The lentivirus vector of claim 14, wherein the promoter is human cytomegalovirus (CMV) immediate early promoter, EFl -alpha promoter, or promoter of UBC.

16. The lentivirus vector of claim 13, wherein the promoter directs expression of the SPG11 gene at least in neurons.

17. The lentivirus vector of claim 16, wherein the promoter is a neuron-specific promoter, and which is optionally a synapsin I promoter, CamKII promoter, MeCP2 promoter, NSE promoter, and Hb9 promoter, or a derivative thereof.

18. The lentivirus vector of claim 16, wherein the promoter directs expression in glial cells, oligodendrocytes, and / or astrocytes.

19. The lentivirus vector of claim 18, wherein the promoter is selected from MBP promoter, PLP1 promoter, and CNP promoter, GFAP promoter, SI 00b promoter, and a derivative thereof.

20. The lentivirus vector of any one of claims 1 to 12, wherein the promoter is the SPG11 endogenous promoter or core promoter elements thereof.

21. The lentivirus vector of claim 20, wherein the promoter comprises at least 200 bps, or at least 300 bps, or at least 500 bps, or at least 800 bps, or at least 1000 bps, or at least 1500 bps upstream from the SPG11 transcription start site.

22. The lentivirus vector of any one of claims 1 to 21, wherein the vector is derived from HIV-1 and integrates in actively transcribed genes.

23. The lentivirus vector of claim 22, wherein the lentivirus vector comprises replicationdefective viral particles having a self-inactivating (SIN) 3' LTR.

24. The lentivirus vector of claim 23, wherein lentivirus vector maintains cis-acting viral sequences necessary for encapsidation, reverse transcription, and integration in the host cell genome.

25. The lentivirus vector of any one of claims 22 to 24, wherein the viral particles comprise VSV-G glycoprotein.

26. The lentivirus vector of any one of claims 22 to 25, comprising an SPG11 expression cassette comprising the nucleotide sequence of SEQ ID NO: 5.

27. A method for treating a patient having a condition associated with SPG11 mutation, comprising administering the lentivirus vector of any one of claims 1 to 26.

28. The method of claim 27, wherein the patient exhibits lysosomal dysfunction.

29. The method of claim 27 or 28, wherein the patient has at least one pathogenic SPG11 mutation in one or both alleles, optionally wherein mutations are compound heterozygous mutations.

30. The method of claim 29, wherein pathogenic mutations are in SPG11 introns or exons.

31. The method of claim 30, wherein at least one pathogenic mutation is in exon 1, 3, 4, 6, 7, 8, 10, 11, 12, 15, 16, 25, 30, 31, 32, 34, 36, or 37 of SPG11.

32. The method of claim 30 or 31, wherein the pathogenic mutation is a loss-of-function mutation, and may cause one or more of a frameshift, truncation, an amino acid change, and deletion or insertion of one or more amino acids.

33. The method of any one of claims 27 to 32, wherein the patient has hereditary spastic paraplegia (HSP), Kjellin syndrome juvenile amyotrophic lateral sclerosis, or parkinsonism.

34. The method of claim 33, wherein the patient exhibits thinning of the corpus callosum.

35. The method of any one of claims 27 to 34, wherein the patient does not show any signs of clinical symptoms, but carries pathogenic mutations in both SPG11 genes.

36. The method of any one of claims 27 to 35, wherein the patient is a pediatric or neonatal patient.

37. The method of any one of claims 27 to 35, the patient is an adult or adolescent patient.

38. The method of any one of claims 27 to 37, wherein vector is administered by systemic delivery.

39. The method of any one of claims 27 to 37, wherein the vector is administered to the brain or CNS.

40. The method of any one of claims 27 to 39, wherein the vector is administered at a multiplicity of infection (MOI) selected from the ranges of MOIs comprising 0.5 to 20, 0.5 to 15, 0.5 to 14, 0.5 to 13, 0.5 to 12, 0.5 to 11, 0.5 to 10, and 1 to 10.

41. The method of any one of claims 27 to 39, wherein the vector is administered at an MOI less than 10.

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