Regulatory elements for targeted expression of cargo genes in spinal cord cells

Abstract

Methods and materials for expressing one or more desired polypeptides in a specific type, subtype, or population of cells are provided herein. For example, this document provides nucleic acids that contain a cell population-specific RE coupled to a transgene, as well as methods for using such nucleic acids to drive expression of transgenes in selected populations of neurons within the spinal cord dorsal horn.

Description

[0001] REGULATORY ELEMENTS FOR TARGETED EXPRESSION OF CARGO GENES IN SPINAL CORD CELLS

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims benefit of priority from U.S. Provisional Application No. 63 / 558,493, filed on February 27, 2024. The disclosure of the prior application is considered part of (and is incorporated by reference in) the disclosure of this application.

[0004] STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

[0005] This invention was made with government support under NS 107364, NS 133364, and NS 109792 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] SEQUENCE LISTING

[0007] This application contains a Sequence Listing that has been submitted electronically as an XML file named “21600-0147W01_SL_ST26.XML.” The XML file, created on February 26, 2025, is 141,690 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety.

[0008] TECHNICAL FIELD

[0009] This document relates to methods and materials for expressing one or more desired polypeptides in particular populations of cells. For example, this document relates to nucleic acids that contain a cell population-specific RE coupled to a transgene, and to methods for using such nucleic acids to drive or enhance expression of transgenes in specific cell populations of the dorsal horn.

[0010] BACKGROUND

[0011] Chronic pain is a common and debilitating medical condition and is a critical driver of the opioid epidemic in the United States. Existing medical options for chronic pain are lacking in efficacy and safety (Cohen et al., Lancet, 397:2082-2097, 2021). The most effective class of analgesics, opioids, has precipitated a catastrophic and ongoing epidemic of morbidity and mortality (Skolnick, Annu Rev Pharmacol Toxicol, 58:143- 149, 2018). Analgesics also can have significant off-target effects, such as severe constipation, which can lead to tolerance and withdrawal (Jamison and Mao, Mayo Clin Proc, 09:957-968, 2015). Local treatments, such as epidurals, also can have off-target hemodynamic and neurological effects (Youssef et al., Anesthesia & Analgesia, 119:965- 977, 2014).

[0012] Signals of bodily pain are transmitted by neurons in the dorsal horn of the spinal cord before traveling to the brain (Todd, Nat Rev Neurosci, 11:823-836, 2010; and Piers and Seal, Science, 354:578-584, 2016). The dorsal horn is thus a major determinant of pain perception. However, there is currently no means to specifically target the dorsal horn neurons involved in pain perception without affecting the normal activity of spinal cord neurons involved in functions other than pain, such as movement and sensation.

[0013] SUMMARY

[0014] This document is based, at least in part, on the development of biological tools that can be used to treat chronic pain and other neurological disorders. For example, this document provides viruses containing cell population-specific REs to drive expression of a transgene in specific cell populations of the dorsal horn that are important for pain. In some cases, the transgene can encode a receptor that can activate (in inhibitory neurons) or inhibit (in excitatory neurons) neuronal activity. The cell population-specific REs can allow for the targeted expression of the transgenes delivered by the virus. These elements are small and can fit within the virus packaging limits. In addition, the specificity helps to avoid off target side effects.

[0015] As demonstrated herein, a machine learning-based approach was used that leverages the cell population specificity of REs to control gene expression in specific neuronal cell population in the brain (Lawler et al., eLife, ll:e69571, 2022). In this paradigm, termed Specific Nuclear- Anchored Independent Labeling (SNAIL), deep neural networks were used to identify candidate regulatory element (RE) sequences with highly specific activity in neuronal cell populations of interest. Candidate REs were then synthesized and packaged into an engineered adeno-associated virus (AAV) along with a transgene (e.g., a reporter gene such as GFP or a protein that controls neuronal firing) to be expressed in target cells.

[0016] In a first aspect, this document features a nucleic acid construct that includes (a) a nucleotide sequence encoding a polypeptide, and (b) at least one RE specific for a selected cell population, where the RE includes the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97, or a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS: 24-97, where the RE is operably linked to the sequence encoding the polypeptide and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population. The polypeptide can be a Designer Receptor Exclusively Activated by Designer Drug (DREADD) polypeptide. The DREADD polypeptide can be a hM4Di polypeptide, a hM3Dq polypeptide, a PS AM4-GlyR polypeptide, or a PS AM4- 5HT3 polypeptide. The polypeptide can be a polypeptide that reduces transmitter release from neurons. The nucleic acid can further include a nucleotide sequence encoding a tag polypeptide, such that when the nucleotide sequences encoding the polypeptide and the tag polypeptide are expressed, the polypeptide is coupled to the tag polypeptide. The tag polypeptide can be a fluorescent polypeptide. The fluorescent polypeptide can be a green fluorescent protein (GFP). The GFP can include an amino acid sequence having at least 95% sequence identity with the superfolder GFP sequence set forth in SEQ ID NO:20. The fluorescent polypeptide can be selected from the group consisting of mCherry, mCitrine, mRuby, nuclear localization sequence-GFP, tdTomato, and SunlGFP The nucleic acid construct can further include virus sequences. The virus sequences can be adeno-associated virus (AAV) sequences or lentivirus sequences. The RE can be specific for a population of dorsal horn spinal cord neurons, such that the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons transduced with the nucleic acid construct but is not expressed in at least 90% of other dorsal horn spinal cord neuron populations transduced with the nucleic acid construct. In another aspect, this document features a virus particle containing a nucleic acid construct that includes (a) a nucleotide sequence encoding a polypeptide, and (b) at least one RE specific for a selected cell population, where the RE includes the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24- 97, or a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97, where the RE is operably linked to the sequence encoding the polypeptide and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population. The polypeptide can be a Designer Receptor Exclusively Activated by Designer Drug (DREADD) polypeptide. The DREADD polypeptide can be a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4-5HT3 polypeptide. The polypeptide can be a polypeptide that reduces transmitter release from neurons. The nucleic acid can further include a nucleotide sequence encoding a tag polypeptide, such that when the nucleotide sequences encoding the polypeptide and the tag polypeptide are expressed, the polypeptide is coupled to the tag polypeptide. The tag polypeptide can be a fluorescent polypeptide. The fluorescent polypeptide can be a green fluorescent protein (GFP). The GFP can include an amino acid sequence having at least 95% sequence identity with the superfolder GFP sequence set forth in SEQ ID NO:20. The fluorescent polypeptide can be selected from the group consisting of mCherry, mCitrine, mRuby, nuclear localization sequence-GFP, tdTomato, and SunlGFP The nucleic acid construct can further include virus sequences. The virus sequences can be adeno-associated virus (AAV) sequences or lentivirus sequences. The RE can be specific for a population of dorsal horn spinal cord neurons, such that the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons transduced with the nucleic acid construct but is not expressed in at least 90% of neurons of other dorsal horn spinal cord neuron populations transduced with the nucleic acid construct. The virus can be AAV or lentivirus. In another aspect, this document features a method for modifying a selected cell population within a broader population of cells. The method can include, or consist essentially of, introducing into the broader population of cells a nucleic acid construct, where the nucleic acid construct includes (a) a sequence encoding a polypeptide, and (b) at least one RE specific for the selected cell population, where the RE includes the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97, or a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97, where the RE is operably linked to the sequence encoding the polypeptide, and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population, and where the polypeptide is expressed in the selected cell population. The polypeptide can be a DREADD polypeptide. The polypeptide can be a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4-5HT3 polypeptide. The polypeptide can be a polypeptide that reduces transmitter release from neurons. The nucleic acid can further include a nucleotide sequence encoding a tag polypeptide, such that when the nucleotide sequences encoding the polypeptide and the tag polypeptide are expressed, the polypeptide is coupled to a tag polypeptide. The tag polypeptide can be a fluorescent polypeptide. The fluorescent polypeptide can be a GFP. The GFP can include an amino acid sequence having at least 95% sequence identity with the sequence set forth in SEQ ID NO:20. The fluorescent polypeptide can be selected from the group consisting of mCherry, mCitrine, mRuby, nuclear localization signal-GFP, tdTomato, and SunlGFP The nucleic acid construct can further include virus sequences. The virus sequences can be AAV sequences or lentivirus sequences. The RE can be specific for a population of dorsal horn spinal cord neurons, such that the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced, but is not expressed in at least 90% of dorsal horn spinal cord neurons of other populations into which the nucleic acid construct was introduced. In another aspect, this document features a method for treating pain in a mammal. The method can include, or consist essentially of, administering, to the mammal, a nucleic acid construct that includes (a) a sequence encoding a polypeptide, and (b) at least one RE specific for a selected cell population, where the RE includes the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24- 97, or a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97, where the RE is operably linked to the sequence encoding the polypeptide, and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population, and where the polypeptide is expressed in the selected cell population within the mammal. The mammal can be a human. The polypeptide can be a DREADD polypeptide. The polypeptide can be a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4-5HT3 polypeptide. The polypeptide can be a polypeptide that reduces transmitter release from neurons. The nucleic acid construct can further include virus sequences. The virus sequences can be AAV sequences or lentivirus sequences. The RE can be specific for a population of dorsal horn spinal cord neurons, such that the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced, but is not expressed in at least 90% of dorsal horn spinal cord neurons of other populations into which the nucleic acid construct was introduced.

[0017] In another aspect, this document features a method for treating pain in a mammal, where the method includes, or consists essentially of, administering, to the mammal, a virus containing a nucleic acid that includes (a) a sequence encoding a polypeptide, and (b) at least one RE specific for a selected cell population, where the RE includes the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97, or a nucleotide sequence having at least 95% sequence identity to SEQ ID SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97, where the RE is operably linked to the sequence encoding the polypeptide, and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population, and where the polypeptide is expressed in the selected cell population within the mammal. The mammal can be a human. The polypeptide can be a DREADD polypeptide. The polypeptide can be a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4-5HT3 polypeptide. The polypeptide can be a polypeptide that reduces transmitter release from neurons. The virus can be an AAV or a lentivirus. The RE can be specific for a population of dorsal horn spinal cord neurons, such that the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons to which the virus was administered, but is not expressed in at least 90% of dorsal horn spinal cord neurons of other populations to which the virus was administered.

[0018] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0019] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0020] DESCRIPTION OF DRAWINGS

[0021] FIG. 1 is a schematic illustrating the steps in a representative process for therapeutic development and testing in vivo. The illustrated process includes designing cross-species REs using machine learning to drive expression in target cell populations for pre-clinical testing in vivo.

[0022] FIG. 2A is a schematic showing theorized roles of dorsal horn neuronal subtypes and populations in pain circuitry, along with their corresponding marker genes. “Inflammatory,” “neuropathic,” and “polyneuropathic” refer to the different mechanistic causes of mechanical allodynia. CALB2 are important for inflammatory injuries, PKCy are important for neuropathic injuries, and CCK are important for both inflammatory and neuropathic injuries. GRPR are important for all three mechanistic causes of injuries. FIG. 2B is a schematic showing a low-dimensional representation (UMAP) of macaque snATAC-seq data, indicating well-delineated clusters of the neuronal populations previously identified in snRNA, and demonstrating that these neuronal populations can be studied using open chromatin and cell-type-specific REs. FIG. 2C is a UMAP plot showing that several neuronal populations identified in a macaque snRNA-seq analysis are significantly enriched in chronic pain syndromes, based on stratified LD score regression. Abbreviations: CLPC = chronic localized pain condition; COPC = chronic overlapping pain condition; NumCPsites = number of chronic pain sites; LTMR = light touch mechanoreceptors.

[0023] FIGS. 3A-3I show the results of expression of GFP in dorsal horn neurons transduced by AAV PHP. eb GLUT11.7 EGFP, and show functional data showing the result of activating hM4Di on mechanical thresholds in a persistent pain model. FIG. 3A is a map of an AAV plasmid containing GLUT11.7 RE, EGFP, WPRE and hGH polyA signal in tandem. This construct was packaged into AAV PHP. eb. FIG. 3B is a representative image of an immunohistochemistry stain of EGFP and PAX2 (a marker of inhibitory neurons) in the lumbar dorsal horn of mice injected with AAV PHP. eb GLUT 11.7 EGFP. FIG. 3C is a diagram of a vector for AAV packaging of GLUT 11.7 driving expression of hM4Di-GFP in mammalian dorsal horn. FIG. 3D shows a representative image of GFP (which fluoresced green) and PAX2 (which fluoresced red) expressed in mouse dorsal horn using the AAV PHP. eb GLUT11.7 hM4Di-GFP vector. FIG. 3E is a graph plotting the results of testing von Frey thresholds in adult mice injected with AAV PHP. eb GLUT11.7 hM4Di-GFP in the dorsal horn. Mechanical thresholds were tested on the plantar hind paw before and after injection of complete Freund’s adjuvant (CFA) with saline or clozapine-n-oxide (CNO). FIG. 3F is a map of an AAV plasmid containing GLUT 11.7 RE and hM4Di in tandem, separated from an EGFP coding sequence by a T2A self-cleaving sequence. FIG. 3G is a representative image showing immunohistochemistry staining of EGFP (which fluoresced green) and PAX2 (which fluoresced red) in the lumbar dorsal horn of mice injected with the pAAV GLUT11.7 hM4Di.T2A.EGFP vector. FIG. 3H includes a pair of graphs plotting the 50% paw withdrawal threshold (left panel) and paw withdrawal frequency (right panel) in mice that were injected with AAV8 GLUT11.7 hM4Di T2A EGFP in the dorsal horn and the subjected to mechanical threshold testing on the plantar hind paw, before and 9 weeks after intraperitoneal injection of streptozotocin to induce a model of diabetic neuropathic pain. Saline or clozapine-n-oxide (CNO; a synthetic drug that serves as a ligand to activate the hM4Di receptor) was injected 20 minutes prior to mechanical testing. FIG. 31 includes a pair of graphs plotting the 50% paw withdrawal threshold (left panel) and paw withdrawal frequency (right panel) in mice that were injected with AAV8 GLUT11.7 hM4Di T2A EGFP in the dorsal horn and then subjected to the spared sural nerve injury model. The impact of activating hM4Di on mechanical allodynia was evaluated 1 and 2 weeks after inducing the spared sural nerve injury. Saline or CNO was injected 20 minutes prior to mechanical testing.

[0024] FIGS. 4A-4D show results from expressing GLUT5.92 RE- and GLUT6.97 RE- driven GFP expression in adult C57B1 / 6 mice. FIG. 4A is a map of an AAV plasmid containing GLUT5.92 RE, NLS-sfEGFP, WPRE, and a polyA sequence in tandem. This construct was packaged with AAV PHP. eb. FIG. 4B is a representative image showing immunohistochemistry staining of GFP in the dorsal horn of the spinal cord in mice injected with the virus of FIG. 4A. FIG. 4C is a map of an AAV plasmid containing GLUT6.97 RE, NLS-sfEGFP, WPRE, and a polyA sequence in tandem. This construct was packaged with AAV PHP. eb. FIG. 4D is a representative image showing immunohistochemistry staining of GFP in the dorsal horn of the spinal cord in mice injected with the virus of FIG. 4A. FIG. 5 includes representative nucleic acid and amino acid sequences for hM4Di (SEQ ID NOS: 6 and 7, respectively).

[0025] FIG. 6 includes representative nucleic acid and amino acid sequences for hM3Dq (SEQ ID NOS: 8 and 9, respectively).

[0026] FIG. 7 includes representative nucleic acid and amino acid sequences for PSAM4-GlyR (SEQ ID NOS: 10 and 11, respectively).

[0027] FIG. 8A includes representative nucleic acid and amino acid sequences for PSAM4-HRT5 high conductance (SEQ ID NOS: 12 and 13, respectively). FIG. 8B includes representative nucleic acid and amino acid sequences for PSAM4-HRT5 low conductance (SEQ ID NOS: 14 and 15, respectively).

[0028] FIG. 9 includes representative nucleic acid and amino acid sequences for tetanus toxin light chain (TeTLC) (SEQ ID NOS: 16 and 17, respectively).

[0029] FIG. 10A shows a representative Streptococcus pyogenes Cas9 DNA coding sequence (SEQ ID NO: 18), and FIG. 10B shows a representative S. pyogenes Cas9 amino acid sequence (SEQ ID NO: 19).

[0030] FIGS. 11A-11H include signal tracks of open chromatin of open chromatin in distinct spinal cord cell populations across genomic positions. The degree of open chromatin, its cell population-specificity, and the identity of the nearby genes are considered in prioritizing the regions as candidate REs. FIG. 11 A, GAB A3; FIG. 11B, GABA4; FIG. 11C, GLUT2; FIG. 11D, GLUT5; FIG. HE, GLUT2; FIG. 11F, GLUT11. FIG. 11G, GLUT5; FIG. HH, GLUT6.

[0031] DETAILED DESCRIPTION

[0032] This document provides methods and materials that can be used as tools to, for example, treat chronic pain and other neurological disorders. For example, this document provides nucleic acid constructs containing cell subtype- or population-specific REs that can drive or enhance expression of a selected polypeptide (e.g., a receptor) in a specific neural cell subtype or population that is involved in pain, as well as methods for using such nucleic acid constructs. When the polypeptide is a receptor, the receptor can activate (in inhibitory neurons) or inhibit (in excitatory neurons) neuronal activity. In some cases, the nucleic acid constructs provided herein are viral constructs that can be used to deliver a nucleic acid containing a sequence encoding a selected polypeptide, where the nucleic acid also contains a cell population-specific RE that is operably linked to the sequence encoding the polypeptide. The cell population specific REs allow for targeted expression of the coding sequences delivered by the virus, and are small enough to fit well within virus packaging limits.

[0033] In general, tissues (e.g., the spinal cord) contain different cell types. For example, neurons are one cell type among several others in the spinal cord. Neurons can be divided into a number of different subtypes based, for example, on their anatomy or their developmental origin. These include, without limitation, coarse-grained neurons, finegrained neurons, excitatory neurons, and inhibitory neurons, among others. As used herein, a “population” of cells refers to subsets of cells that are molecularly distinct, such that they can be defined based on their gene expression and / or epigenetic markers. For example, open chromatin and DNA methylation are epigenetic features that can be used to define particular populations of cells (see, e.g., Luo et al., Cell Genom. 2(3): 100107, 2022; doi: 10.1016 / j.xgen.2022.100107).

[0034] In some cases, this document provides recombinant nucleic acid constructs containing (a) a cell population-specific RE, and (b) a nucleotide sequence encoding a desired polypeptide, where the cell population-specific RE can drive expression of the desired polypeptide in the specific cell population. For example, this document provides nucleic acid constructs containing (a) a RE specific for particular neurons of the dorsal horn (e.g., glutamatergic cell populations (GLUT 1-11) and GABAergic cell populations (GABA1-5); see, e.g., Arokiaraj et al., bioRxiv, 2022.04.01.486135, 2022; now Arokiaraj et al., Cell Reports, 43(11):114876, 2024), which correspond to human populations Ex- 1 - 12 and Inh-1-9 as shown by Yadav et al., Neuron, 11 l(3):328-344.e7, 2023, and (b) a transgene encoding a desired polypeptide, where the RE is operably linked to the transgene and therefore can drive expression of the desired polypeptide in the dorsal horn neurons. For example, dorsal horn neuron populations referred to as GAB A3 and / or GABA5, whose activation can prevent the emergence of mechanical allodynia prior to injury, can be targeted after injury with a construct containing an appropriate RE and encoding a DREADD that can activate GAB A3 and / or GABA5, in order to alleviate mechanical allodynia. Cell populations were defined by single nuclear sequencing of the dorsal horn of the spinal cord of rhesus macaque as described by Arokiaraj et al., supra, followed by cross-species comparisons to dorsal horn single nuclei or cell sequencing studies in mice and humans to identify the analogous cell populations in those species. In some cases, dorsal horn neuron populations GLUT2, GLUT5, GLUT6, GLUT9, GLUT10, and / or GLUT11, which are involved in transmission of mechanical allodynia after injury, can be targeted with a construct containing an appropriate RE and encoding a DREADD that can inhibit one or more of these populations in order to alleviate mechanical allodynia. It is noted that targeted expression of a DREADD in one or more of these cell populations also can be used to alleviate spontaneous pain. In some cases, a recombinant nucleic acid construct provided herein can contain a minimal promoter in addition to the RE and the nucleotide sequence encoding a desired polypeptide.

[0035] The terms “nucleic acid” and “polynucleotide” can be used interchangeably, and refer to both RNAand DNA, including cDNA, genomic DNA, synthetic (e.g., chemically synthesized) DNA, and DNA (or RNA) containing nucleic acid analogs. Polynucleotides can have any three-dimensional structure. A nucleic acid can be double-stranded or single-stranded (i.e., a sense strand or an antisense single strand). Non-limiting examples of polynucleotides include genes, gene fragments, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers, as well as nucleic acid analogs.

[0036] As used herein, “isolated,” when in reference to a nucleic acid, refers to a nucleic acid that is separated from other nucleic acids that are present in a genome, including nucleic acids that normally flank one or both sides of the nucleic acid in the genome. The term “isolated” as used herein with respect to nucleic acids also includes any non- naturally occurring sequence, since such non-naturally occurring sequences are not found in nature and do not have immediately contiguous sequences in a naturally occurring genome. An isolated nucleic acid can be, for example, a DNA molecule, provided one of the nucleic acid sequences normally found immediately flanking that DNA molecule in a naturally occurring genome is removed or absent. Thus, an isolated nucleic acid includes, without limitation, a DNA molecule that exists as a separate molecule (e.g., a chemically synthesized nucleic acid, or a cDNA or genomic DNA fragment produced by PCR or restriction endonuclease treatment) independent of other sequences, as well as DNA that is incorporated into a vector, an autonomously replicating plasmid, a virus (e.g., a pararetrovirus, a retrovirus, lentivirus, adenovirus, or herpes virus), or the genomic DNA of a prokaryote or eukaryote. In addition, an isolated nucleic acid can include a recombinant nucleic acid such as a DNA molecule that is part of a hybrid or fusion nucleic acid. A nucleic acid existing among hundreds to millions of other nucleic acids within, for example, cDNA libraries or genomic libraries, or gel slices containing a genomic DNA restriction digest, is not to be considered an isolated nucleic acid.

[0037] A nucleic acid can be made by, for example, chemical synthesis or polymerase chain reaction (PCR). PCR refers to a procedure or technique in which target nucleic acids are amplified. PCR can be used to amplify specific sequences from DNA as well as RNA, including sequences from total genomic DNA or total cellular RNA. Various PCR methods are described, for example, in PCR Primer: A Laboratory Manual. Dieffenbach and Dveksler, eds., Cold Spring Harbor Laboratory Press, 1995. Generally, sequence information from the ends of the region of interest or beyond is employed to design oligonucleotide primers that are identical or similar in sequence to opposite strands of the template to be amplified. Various PCR strategies also are available by which site-specific nucleotide sequence modifications can be introduced into a template nucleic acid.

[0038] The term “RE” (which may be used interchangeably with the terms “regulatory region,” “control element,” and “expression control sequence”) refers to a nucleotide sequence that influences transcription or translation initiation and rate. REs can include, without limitation, promoter sequences, enhancer sequences, response elements, protein recognition sites, inducible elements, promoter control elements, protein binding sequences, 5' and 3' untranslated regions (UTRs), and / or transcriptional start sites. As used herein, “operably linked” means incorporated into a genetic construct so that expression control sequences effectively control expression of a coding sequence of interest. A coding sequence is “operably linked” and “under the control” of expression control sequences in a cell when RNA polymerase is able to transcribe the coding sequence into RNA, which if an mRNA, then can be translated into the protein encoded by the coding sequence. Thus, a RE can modulate (e.g., enhance, regulate, facilitate, or drive) transcription in the cell in which it is desired to express a selected nucleic acid. For example, a cell specific RE that confers transcription only or predominantly in a particular neuron cell type, subtype, or population can be used. Such REs can be identified using methods such as those described herein and can be used in the nucleic acid constructs provided herein.

[0039] In some cases, a RE described herein can serve as a promoter. In some cases, a regulator element described herein can be used in combination with a promoter (e.g., a minimal promoter that allows for formation of a transcription initiation complex). A promoter is an expression control sequence composed of a region of a DNA molecule, typically (but not always) within 100 nucleotides upstream of the point at which transcription starts (generally near the initiation site for RNA polymerase II). Promoters are involved in recognition and binding of RNA polymerase and other to initiate and modulate transcription. To bring a coding sequence under the control of a promoter, it typically is necessary to position the translation initiation site of the translational reading frame of the polypeptide between one and about fifty nucleotides downstream of the promoter. A promoter can, however, be positioned as much as about 5,000 nucleotides upstream of the translation start site, or about 2,000 nucleotides upstream of the transcription start site. A promoter typically includes at least a core (basal) promoter. A promoter also may include at least one control element such as an upstream element. Such elements include upstream activation regions (UARs) and, optionally, other DNA sequences that affect transcription of a polynucleotide such as a synthetic upstream element.

[0040] Any appropriate cell population specific RE can be included in the nucleic acid constructs provided herein. An RE can have any appropriate length. For example, an RE can have a length from about 300 nucleotides to about 700 nucleotides (e.g., from about 300 to about 350, from about 350 to about 400, from about 400 to about 450, from about 450 to about 500, from about 450 to about 550, from about 550 to about 600, from about 600 to about 650, or from about 650 to about 700 nucleotides).

[0041] In some cases, a cell population specific RE can be effective to drive expression of a coding sequence to which it is operably linked in dorsal horn neurons. Non-limiting examples of suitable REs are set forth in SEQ ID NOS: 1-5 and 21-97 herein. For example, a RE can have the sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97. In some cases, a RE can have a sequence that is at least 75% identical (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97. For example, a RE can have a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO: 1, at least 90% identical to the nucleotide sequence set forth in SEQ ID NO:2, at least 90% identical to the nucleotide sequence set forth in SEQ ID NO: 3, at least 90% identical to the nucleotide sequence set forth in SEQ ID NO:4, at least 90% identical to the nucleotide sequence set forth in SEQ ID NO: 5, at least 90% identical to the nucleotide sequence set forth in SEQ ID NO:21, at least 90% identical to the nucleotide sequence set forth in SEQ ID NO:22, at least 90% identical to the nucleotide sequence set forth in SEQ ID NO: 23, or at least 90% identical to the nucleotide sequence set forth in any of SEQ ID NOS: 24-97. In some cases, a RE can have a nucleotide sequence that is at least 95% identical to the nucleotide sequence set forth in SEQ ID NO: 1, at least 95% identical to the nucleotide sequence set forth in SEQ ID NO:2, at least 95% identical to the nucleotide sequence set forth in SEQ ID NO: 3, at least 95% identical to the nucleotide sequence set forth in SEQ ID NO:4, at least 95% identical to the nucleotide sequence set forth in SEQ ID NO: 5, at least 95% identical to the nucleotide sequence set forth in SEQ ID NO:21, at least 95% identical to the nucleotide sequence set forth in SEQ ID NO:22, at least 95% identical to the nucleotide sequence set forth in SEQ ID NO:23, or at least 95% identical to the nucleotide sequence set forth in any of SEQ ID NOS: 24-97.

[0042] The percent sequence identity between a particular nucleic acid or amino acid sequence and a nucleic acid or amino acid sequence referenced by a particular sequence identification number is determined as follows. First, a nucleic acid or amino acid sequence is compared to the sequence set forth in a particular sequence identification number using the BLAST 2 Sequences (B12seq) program from the stand-alone version of BLASTZ containing BLASTN version 2.0.14 and BLASTP version 2.0.14. This standalone version of BLASTZ can be obtained from Fish & Richardson’s web site (e.g., www.fr.com / blast / ) or the U.S. government’s National Center for Biotechnology Information web site (www.ncbi.nlm.nih.gov). Instructions explaining how to use the B12seq program can be found in the readme file accompanying BLASTZ. B12seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. To compare two nucleic acid sequences, the options are set as follows: -i is set to a file containing the first nucleic acid sequence to be compared (e.g., C:\seql.txt); -j is set to a file containing the second nucleic acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastn; -o is set to any desired file name (e.g., C:\output.txt); -q is set to -1; -r is set to 2; and all other options are left at their default setting. For example, the following command can be used to generate an output file containing a comparison between two sequences: C:\B12seq -i c:\seql.txt -j c:\seq2.txt -p blastn -o c:\output.txt -q -1 -r 2. To compare two amino acid sequences, the options of B12seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C:\seql.txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C:\output.txt); and all other options are left at their default setting. For example, the following command can be used to generate an output file containing a comparison between two amino acid sequences: C:\B12seq -i c:\seql.txt -j c:\seq2.txt -p blastp -o c:\output.txt. If the two compared sequences share homology, then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology, then the designated output file will not present aligned sequences.

[0043] Once aligned, the number of matches is determined by counting the number of positions where an identical nucleotide or amino acid residue is presented in both sequences. A matched position refers to a position in which an identical nucleotide or amino acid residue occurs at the same position in aligned sequences. The percent sequence identity is determined by dividing the number of matches by the length of the sequence set forth in the identified sequence (e.g., SEQ ID NO: 1), followed by multiplying the resulting value by 100. For example, an amino acid sequence that has 480 matches when aligned with the sequence set forth in SEQ ID NO: 1 is 95.8 percent identical to the sequence set forth in SEQ ID NO: 1 (i.e., 480 501 x 100 = 95.8). It is noted that the percent sequence identity value is rounded to the nearest tenth. For example, 75.11, 75.12, 75.13, and 75.14 are rounded down to 75.1, while 75.15, 75.16, 75.17, 75.18, and 75.19 are rounded up to 75.2. It also is noted that the length value will always be an integer.

[0044] Any appropriate method can be used to identify cell population-specific REs for inclusion in the nucleic acid constructs provided herein. As described in Example 1 herein, for example, Specific Nuclear- Anchored Independent Labeling (SNAIL) methodology was then used to identify several REs as being likely to selectively target dorsal horn neurons. SNAIL methodology is described in detail in, for example, WO 2020 / 257520 (see, e.g., pages 4-6 and 21-26). In addition to a cell population specific RE, the nucleic acid constructs described herein can contain a nucleotide sequence encoding a desired polypeptide. The term “polypeptide” as used herein refers to a compound of two or more subunit amino acids, regardless of post- translational modification (e.g., phosphorylation or glycosylation). The subunits may be linked by peptide bonds or other bonds such as, for example, ester or ether bonds. The term “amino acid” refers to either natural and / or unnatural or synthetic amino acids, including D / L optical isomers.

[0045] A nucleic acid construct provided herein can contain any appropriate coding sequence for a polypeptide that is to be expressed in particular cells (e.g., dorsal horn neurons, or a particular subtype or population of dorsal horn neurons). In some cases, a polypeptide encoded by a nucleic acid construct provided herein can be a Designer Receptor Exclusively Activated by Designer Drug (DREADD) polypeptide. DREADD polypeptides are a class of artificially engineered protein receptors that can be selectively activated by certain ligands. Examples of DREADD polypeptides include, without limitation, hM4Di, hM3Dq, PSAM4-GlyR, and PSAM4-5HT3. For example, the PSAM4 DREADDs are fusion proteins that include a mutated ligand binding domain from the human nicotinic alpha 7 receptor and an ion channel domain from the human glycine receptor (GlyR) or the human 5HT3 receptor (which has high conductance and low conductance forms). The mutations (e.g., Y115F, Q79R, Q139G, Q139V, Q139W, Q139Y, L141A, L141Q, and / or LI 4 IS) in the ligand binding domain can decrease the binding affinity of the endogenous ligand acetylcholine. See, e.g., Magnus et al., Science, 364(6436):eaav5282, 2019; and Sternson and Roth, Annu Rev Neurosci., 37:387-407, 2014. The PSAM4 DREADDs bind with relatively high affinity and selectivity to varenicline (an FDA approved smoking cessation aid that can block the effects of nicotine in the brain), as well as to novel ligands described elsewhere (see, e.g., Magnus et al., supra). hM3Dq and hM4Di are forms of human muscarinic receptors that have been mutated to have a much lower affinity for acetylcholine and a higher affinity for clozapine and related compounds. hM3Dl and hM4Di are second messenger coupled receptors that increase or decrease the membrane potential, respectively, by causing the modification of ion channels, which results in the activation or inhibition, respectively, of neurotransmitter release. See, e.g., Armbruster et al., Proc Natl Acad Sci USA, 104(12):5163-5168, 2007; Roth, Neuron, 89:694-693, 2016; and Gomez et al., Science, 357(6350):503-507, 2017.

[0046] An exemplary nucleotide sequence encoding a hM4Di polypeptide is set forth in SEQ ID NO: 6, and an exemplary hM4Di amino acid sequence is set forth in SEQ ID NO:7 (FIG. 5). In some cases, a polypeptide coding sequence in a nucleic acid construct provided herein can have a nucleotide sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the nucleotide sequence set forth in SEQ ID NO:6. In some cases, a polypeptide encoded by a nucleic acid construct provided herein can have an amino acid sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence set forth in SEQ ID NO: 7.

[0047] An exemplary nucleotide sequence encoding a hM3Dq polypeptide is set forth in SEQ ID NO: 8, and an exemplary hM3Dq amino acid sequence is set forth in SEQ ID NO:9 (FIG. 6). In some cases, a polypeptide coding sequence in a nucleic acid construct provided herein can have a nucleotide sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the nucleotide sequence set forth in SEQ ID NO: 8. In some cases, a polypeptide encoded by a nucleic acid construct provided herein can have an amino acid sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence set forth in SEQ ID NO: 9.

[0048] An exemplary nucleotide sequence encoding a PS AM4-GlyR polypeptide is set forth in SEQ ID NO: 10, and an exemplary PSAM4-GlyR amino acid sequence is set forth in SEQ ID NO: 11 (FIG. 7). In some cases, a polypeptide coding sequence in a nucleic acid construct provided herein can have a nucleotide sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the nucleotide sequence set forth in SEQ ID NO: 10. In some cases, a polypeptide encoded by a nucleic acid construct provided herein can have an amino acid sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence set forth in SEQ ID NO:11.

[0049] An exemplary nucleotide sequence encoding a PSAM4-5HT3 high conductance (HC) polypeptide is set forth in SEQ ID NO: 12, and an exemplary PSAM4-5HT3 HC amino acid sequence is set forth in SEQ ID NO: 13 (FIG. 8A). An exemplary nucleotide sequence encoding a PSAM4-5HT3 low conductance (LC) polypeptide is set forth in SEQ ID NO: 14, and an exemplary PSAM4-5HT3 amino acid sequence is set forth in SEQ ID NO: 15 (FIG. 8B). In some cases, a polypeptide coding sequence in a nucleic acid construct provided herein can have a nucleotide sequence that is at least 90% (e.g., at least at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, 96%, at least 97%, at least 98%, or at least 99%) identical to the nucleotide sequence set forth in SEQ ID NO: 12. In some cases, a polypeptide encoded by a nucleic acid construct provided herein can have an amino acid sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence set forth in SEQ ID NO: 13. In some cases, a polypeptide coding sequence in a nucleic acid construct provided herein can have a nucleotide sequence that is at least 90% (e.g., at least at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, 96%, at least 97%, at least 98%, or at least 99%) identical to the nucleotide sequence set forth in SEQ ID NO: 14. In some cases, a polypeptide encoded by a nucleic acid construct provided herein can have an amino acid sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence set forth in SEQ ID NO: 15.

[0050] In some cases, a polypeptide encoded by a nucleic acid construct provided herein can be a polypeptide that reduces (e.g., blocks) transmitter release from neurons. Examples of polypeptides that can block transmitter release from neurons include, without limitation, hM4Di, PSAM4-GlyR, and tetanus toxin light chain (TeTLC). Exemplary nucleotide coding sequences and amino acid sequences for hM4Di are set forth in SEQ ID NOS: 6 and 7 above, and exemplary nucleotide coding sequences and amino acid sequences for PSAM4-GlyR are set forth in SEQ ID NOS: 10 and 11 above. Exemplary nucleotide coding sequences and amino acid sequences for TeTLC are set forth in SEQ ID NOS: 16 and 17, respectively (FIG. 9). In some cases, a polypeptide coding sequence in a nucleic acid construct provided herein can have a nucleotide sequence that is at least 90% (e.g., at least at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, 96%, at least 97%, at least 98%, or at least 99%) identical to the nucleotide sequence set forth in SEQ ID NO: 16. In some cases, a polypeptide encoded by a nucleic acid construct provided herein can have an amino acid sequence that is at least 90% (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence set forth in SEQ ID NO:17.

[0051] Polypeptides such as hM4Di, PSAM4-GlyR, and TeTLC can be effective reduce transmitter release from neurons by at least 10% (e.g., at least 20%, at least 25%, at least 40%, at least 50%, at least 75%, at least 90%, at least 95%, at least 99%, or 100%), as compared to the level of transmitter release prior to exposure of the neurons to the polypeptide. Transmitter release from neurons can be measured by any appropriate method. For example, transmitter release from neurons can be measured by assess transmitter release by electrophysiological recording in spinal cord slices.

[0052] In some cases, a polypeptide encoded by a nucleic acid construct provided herein can be a clustered regularly interspaced short palindromic repeats- (CRISPR-) associated (Cas) nuclease. The CRISPR / Cas system includes components of a prokaryotic adaptive immune system that is functionally analogous to eukaryotic RNA interference, using RNA base pairing to direct DNA or RNA cleavage. The Cas protein functions as an endonuclease, and CRISPR RNA (crRNA) and tracer RNA (tracrRNA) sequences complex with the Cas enzyme and direct it to a target DNA sequence. See, e.g., Makarova et al., Nat Rev Microbiol 9(6):467-477, 2011. In some cases, crRNA and tracrRNA can be engineered as a single cr / tracrRNA hybrid (also referred to as a “guide RNA” or “gRNA”) to direct Cas9 cleavage activity (see, e.g., Jinek et al., Science, 337(6096):816-821, 2012). The combination of Cas, crRNA, and tracrRNA (or Cas and gRNA) can then cleave linear or circular dsDNA targets that are complementary to a spacer within the CRISPR cluster. By pairing an RE for a particular subtype or population of neurons with CRISPR / Cas components targeted to a particular gene, genes within the selected subtype or population of neurons can be edited or inactivated when the RE drives or enhances expression of the CRISPR RNAs and the Cas nuclease in the selected neurons. For example, the vesicular glutamate transporter 2 gene (VLGUT2) in excitatory neurons can be targeted using (a) an RE that promotes or enhances expression in excitatory neurons in combination with (b) CRISPR RNA targeted to one or more VGLUT2 sequences. The homology region within the crRNA sequence (the sequence that targets the crRNA to a desired DNA sequence) can be from about 10 to about 40 (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40) nucleotides in length. The tracrRNA hybridizing region within each crRNA sequence can be from about 8 to about 20 (e.g., 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) nucleotides in length. The overall length of a crRNA sequence can be, for example, from about 20 to about 80 (e.g., 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80) nucleotides, while the overall length of a tracrRNA can be, for example, from about 10 to about 30 (e.g., 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, or 30) nucleotides. The overall length of a gRNA sequence, which includes a homology region and a stem loop region that contains a crRNA / tracrRNA hybridizing region and a linker-loop sequence, can be from about 30 to about 110 (e.g., 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, or 130) nucleotides.

[0053] A CRISPR / Cas system can be targeted to any other appropriate gene within any appropriate subtype or population of neurons. For example, a CRISPR / Cas system can be targeted to a gene whose disruption would prevent transmitter release from an excitatory neuron, and / or to a gene whose disruption would increase transmitter release from an inhibitory neuron.

[0054] A representative nucleic acid sequence encoding a Cas polypeptide (in particular, a Cas9 polypeptide from Streptococcus pyogenes) is set forth in SEQ ID NO: 18, and a representative Cas9 amino acid sequence is set forth in SEQ ID NO: 19 (FIGS. 10A and 10B). See, also, NCBI Ref. NC_017053.1 and GENBANK® accession no. AKP81606.1 for the nucleotide and amino acid sequences, respectively. It is to be noted, however, that there are multiple types of Cas nucleases (e.g., Casl2a, Cas3, and CaslO) that can be used in the methods provided herein.

[0055] The polypeptides encoded by the nucleic acid constructs provided herein can have any appropriate length. For example, a polypeptide encoded by a nucleic acid construct provided herein can be between about 20 amino acids and about 750 amino acids (e.g., about 20 to 50 amino acids, about 50 to about 100 amino acids, about 100 to about 200 amino acids, about 200 to about 400 amino acids, about 400 to about 600 amino acids, or about 600 to about 750 amino acids) in length.

[0056] In some cases, the nucleic acid constructs provided herein can contain sequences encoding one or more additional components. For example, a nucleic acid construct provided herein can include a nucleotide sequence encoding a polypeptide tag. In such cases, the nucleotide sequence encoding the polypeptide tag can be linked to the polypeptide coding sequence, such that expression of the polypeptide and tag coding sequences results in a fusion polypeptide containing the desired polypeptide coupled to the tag. The inclusion of a tag can, for example, enable cells and / or nuclei containing the expressed fusion polypeptide to be identified and isolated away from cells and / or nuclei that do not express the fusion polypeptide.

[0057] Any appropriate polypeptide tag can be encoded by the nucleic acid constructs provided herein. In some cases, a polypeptide tag can be a fluorescent polypeptide. For example, a nucleic acid construct can include a nucleotide sequence encoding green fluorescent protein (GFP) (e.g., a superfolder GFP, such as the superfolder GFP having the amino acid sequence set forth in SEQ ID NO:20 or having an amino acid sequence at least 95% identical to the amino acid sequence set forth in SEQ ID NO:20). MRKGEELFTGWPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLP VPWPTLVTTLTYGVQCFARYPDHMI<QHDFFI<SAMPEGYVQERTISFI<DDGTYI< TRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK ANFI<IRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSI<DPNEI<RD HMVLLEFVTAAGITHGMDELYK (SEQ ID NO: 20)

[0058] Other examples of fluorescent polypeptide tags include, without limitation, mCherry, mCitrine, m-Ruby, nuclear localization sequence-GFP, tdTomato, and SunlGFP.

[0059] In some cases, a recombinant nucleic acid provided herein can integrate into the genome of a cell. For example, a recombinant nucleic acid provided herein can integrate into the genome of a cell via illegitimate (random, non-homologous, non-site-specific) recombination. In some cases, a recombinant nucleic acid provided herein can be designed to integrate into the genome of a cell via homologous recombination. Nucleic acid sequences designed for integration via homologous recombination can be flanked on both sides with sequences that are similar or identical to endogenous target nucleotide sequences, which can facilitate integration of the recombinant nucleic acid at the particular site(s) in the genome containing the endogenous target nucleotide sequences. In some cases, nucleic acid sequences adapted for integration via homologous recombination also can include a recognition site for a sequence-specific nuclease. Alternatively, the recognition site for a sequence-specific nuclease can be located in the genome of the cell to be transformed.

[0060] In some cases, a nucleic acid construct can be included in a vector suitable for transformation of cells. Recombinant vectors can be made using, for example, standard recombinant DNA techniques (see, e.g., Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY). This document also provides recombinant nucleic acid constructs (e.g., vectors) containing the REs and polypeptide coding sequences described herein. A “vector” is a replicon, such as a plasmid, phage, or cosmid, into which another DNA segment may be inserted so as to bring about the replication of the inserted segment. Vector backbones include, for example, plasmids, viruses, artificial chromosomes, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), and phage artificial chromosomes (PACs), as well as RNA vectors, and linear or circular DNA or RNA molecules that include chromosomal, non- chromosomal, semi-synthetic, or synthetic nucleic acids. Vectors include those capable of autonomous replication (episomal vectors) and / or expression of nucleic acids to which they are linked (expression vectors). Generally, a vector is capable of replication when associated with the proper control elements. The term “vector” includes cloning and expression vectors, as well as viral vectors and integrating vectors. An “expression vector” is a vector that includes one or more expression control sequences to control and regulate the transcription and / or translation of another DNA sequence. Suitable expression vectors include, without limitation, plasmids and viral vectors derived from, for example, bacteriophage, baculoviruses, tobacco mosaic virus, herpes viruses, cytomegalovirus, retroviruses, vaccinia viruses, adenoviruses, and adeno-associated viruses. Numerous vectors and expression systems are commercially available. Viral vectors include, without limitation, retrovirus (e.g., lentivirus), adenovirus, parvovirus (e.g., adeno associated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e.g., influenza virus), rhabdo virus (e.g., rabies and vesicular stomatitis virus), paramyxovirus (e.g., measles and Sendai), positive strand RNA viruses such as picornavirus and alphavirus, and double-stranded DNA viruses including adenovirus, herpesvirus (e.g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (e.g., vaccinia, fowlpox and canarypox). Other viruses include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, and hepatitis virus, for example. Examples of retroviruses include avian leukosis-sarcoma, mammalian C-type, B-type viruses, D type viruses, HTLV-BLV group, lentivirus, spumavirus (Coffin, “Retroviridae: The viruses and their replication,” in Fundamental Virology, Third Edition, B. N. Fields, et al., eds., Lippincott-Raven Publishers, Philadelphia, 1996).

[0061] Without being bound by any particular mechanism of action, viral delivery of the nucleic acid constructs provided herein can provide flexibility across cell population and species. When using a virus (e.g., adeno-associated virus or lentivirus) delivery method, the nucleic acid constructs can be introduced into any appropriate mammals (e.g., humans, non-human primates, mice, rats, sheep, pigs, or dogs) through intravenous injection, direct injection into the brain, or any other appropriate method. This delivery procedure can provide a time and resource-efficient way to introduce nucleic acids into particular cell populations, particularly when compared with the intricacies of transgenic breeding. The method also can minimize the number of collateral animals that are bred for an experiment but cannot be used due to undesirable genotypes.

[0062] This document also provides methods for using the nucleic acid constructs described herein to label and isolate nuclei and / or cells of a select type, subtype, or population. The methods can include, for example providing a nucleic acid construct described herein, introducing the construct into cells, and culturing or incubating the cells under conditions in which a polypeptide encoded by the construct is expressed in a selected cell population (and is not expressed in cells that are not of the selected population). In some cases (e.g., when the encoded polypeptide is fused to a tag), expression of the polypeptide can result in labeling of the selected cell population. It is to be noted that in some cases, the population of cells is within a mammal.

[0063] Any appropriate method can be used to introduce a nucleic acid construct provided herein into a population of cells. As used herein, “transformed” and “transfected” encompass the introduction of one or more nucleic acid molecules (e.g., one or more expression vectors) into a cell by any of a number of techniques. Suitable methods for transforming and transfecting host cells can be found, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual (2nd edition), Cold Spring Harbor Laboratory, New York (1989). For example, calcium phosphate precipitation, electroporation, heat shock, lipofection, micro injection, and virus-mediated nucleic acid transfer can be used introduce nucleic acid molecules into cells. In addition, naked DNA can be delivered directly to cells in vivo (see, e.g., U.S. Patent Nos. 5,580,859 and 5,589,466). The isolated nucleic acid molecule transformed into a host cell can be integrated into the genome of the cell or maintained in an episomal state. Thus, host cells can be stably or transiently transfected with a construct containing an isolated nucleic acid molecule provided herein. In some cases, one or more nucleic acid constructs can be incorporated into virus particles (e.g., AAV or lentivirus particles), and the virus particles can be delivered to cells in order to transfer their nucleic acid contents to the cells. When a nucleic acid construct is introduced into a population of cells within a mammal, the construct can be delivered into the cells by intrathecal injection, systemic transduction, or intraspinal injection.

[0064] This document also provides methods for modifying a selected cell type, subtype, or population (e.g., dorsal horn neurons) within a broader population of cells (e.g., spinal cord neurons). A method can include, for example, introducing a nucleic acid construct provided herein into a broader population of cells that includes different cell types, subtypes, or populations. The nucleic acid construct can include a sequence encoding a desired polypeptide, and at least one RE (e.g., one, two, three, or more than three REs) that is operably linked to the sequence encoding the polypeptide and is specific for a particular type, subtype, or population of cells within the broader population. For example, the RE can be effective to drive expression of the sequence encoding the polypeptide in dorsal horn neurons, and the RE can be specific for dorsal horn spinal cord neurons. The use of a RE specific for dorsal horn spinal cord neurons means that the sequence encoding the polypeptide is expressed in a majority of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced but is not expressed in at least 90% (e.g., at least 95%, at least 97%, at least 98%, or at least 99%) of other cells into which the nucleic acid construct was introduced. In another example, an RE can be effective to drive expression of the sequence encoding the polypeptide in a selected population of dorsal horn neurons, and the RE can be specific for the selected population of dorsal horn spinal cord neurons. The use of a RE specific for a particular population of dorsal horn spinal cord neurons means that the sequence encoding the polypeptide is expressed in a majority of the selected population of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced but is not expressed in at least 90% (e.g., at least 95%, at least 97%, at least 98%, or at least 99%) of neurons of other populations of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced. In some cases, the RE can include the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97. In some cases, the RE can include a nucleotide sequence that is at least 90% (e.g., at least 95%) identical to the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24- 97.

[0065] In addition, this document provides methods for treating (e.g., reducing or eliminating) pain in a mammal. In some cases, a method provided herein for treating pain can be effective to reduce the level or intensity of pain (e.g., general pain or a particular type of pain, such as neuropathic pain, musculoskeletal pain, mechanical pain, or inflammatory pain) in a mammal by at least 10% (e.g., at least 20%, at least 25%, at least 30%, at least 50%, at least 75%, or at least 90%), as compared to the level or intensity of pain in the mammal prior to initiation of the method. Any appropriate method can be used to assess / quantify the level or intensity of pain. In some cases, a level of pain can be assessed and / or quantified using a verbal rating scale, a numeric rating scale, a visual analogue scale, quantitative sensory testing, facial grimace, and / or machine learning / computer vision. For example, pain can be assessed in non- human primates using veterinary medicine behavioral tools and methods to automate these tools, including computer vision. To facilitate translation of findings across mouse, non-human primates, and human, pain assessments in non-human primates and humans can include measurements of neural physiology. In non-human primates, this may include invasive electrophysiologic recordings. In humans, this would include noninvasive methods such as functional magnetic resonance imaging, functional near-infrared spectroscopy, and electroencephalography, independently and / or combined with quantitative sensory testing methods. In human studies, pain assessments can extend beyond pain intensity to include other aspects of the pain experience, such as unpleasantness, functional impact, and effects on other key domains (e.g., mental health and sleep).

[0066] A method for treating pain in a mammal can include, for example, administering a nucleic acid construct provided herein to a mammal (e.g., a human), where the nucleic acid construct contains a sequence encoding a polypeptide, and at least one RE (e.g., one, two, three, or more than three REs) that is operably linked to the sequence encoding the polypeptide and is specific for a selected cell population. In some cases, the administering can include introducing a virus containing the nucleic acid construct into the mammal. Again, the RE can be effective to drive expression of the sequence encoding the polypeptide in the selected population, such that the polypeptide is expressed in the selected cell population. For example, the selected cell population can be dorsal horn neurons, and the RE can be specific for dorsal horn spinal cord neurons. The use of a RE specific for dorsal horn spinal cord neurons means that the sequence encoding the polypeptide is expressed in a majority of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced but is not expressed in at least 90% (e.g., at least 95%, at least 97%, at least 98%, or at least 99%) of other neurons into which the nucleic acid construct was introduced. In some cases, the RE can include the nucleotide sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS:24-97. In some cases, the RE can include a nucleotide sequence that is at least 90% (e.g., at least 95%) identical to the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or any of SEQ ID NOS: 24-97.

[0067] In some cases, for example, excitatory neuron subtypes that are important for transmission of mechanical allodynia (painful sensation caused by innocuous stimuli such as light touch) can be targeted using methods provided herein in order to prevent the neurons from releasing transmitter. For example, nucleic acid encoding a DREADD that causes inhibition of transmitter release (e.g., hM4Di or PSAM4-GlyR) can be introduced into neurons in a construct that also includes an RE specific for excitatory neurons, such that the RE can drive expression of the DREADD in the excitatory neurons. In some cases, nucleic acid encoding CRISPR / Cas components targeted to VGLUT2 can be introduced into neurons in a construct that also includes an RE specific for excitatory neurons, such that the RE can drive expression of the CRISPR / Cas components in the excitatory neurons, leading to modification or deletion of the VGLUT2 gene so that glutamate cannot be packaged into synaptic vesicles and released from those neurons. In some cases, nucleic acid encoding TeTLC can be introduced into neurons in a construct that also includes an RE for excitatory neurons, such that the RE can drive expression of TeTLC in the excitatory neurons so neurotransmitter cannot be released. In some cases, inhibitory neuron cell subtypes that are important for preventing mechanical allodynia under normal conditions can be targeted. For example, nucleic acid encoding a DREADD that can increase transmitter release (PSAM4-5HT3 or hM3Dq) can be introduced into neurons in a construct that also includes an RE specific for inhibitory neurons, such that the RE can drive expression of the DREADD in the inhibitor neurons.

[0068] A nucleic acid containing a coding sequence operably linked to an RE provided herein (e.g., a nucleic acid encoding a DREADD, where the DREADD encoding sequence is operably linked to any one of SEQ ID NOS: 1 to 5 and 21-97) can be introduced into neurons within a mammal by any appropriate route. In some cases, a nucleic acid can be packaged into an adeno-associated virus (AAV), which can be delivered into neurons within a mammal by, for example, intraspinal or lumbar puncture injection. Any appropriate amount of a nucleic acid or virus can be introduced into a mammal. For example, a composition containing an AAV that contains a nucleic acid provided herein can be injected intraspinally into a mammal in an amount of about 0.1 pL to about 5 pL (e.g., about 0.1 to about 0.5 pL, about 0.5 to about 1 pL, about 1 to about 2 pL, about 2 to about 5 pL, or about 5 to about 10 pL) at a titer of about 1011to about 1015genome copies (GC) per mL (e.g., about 1011to about 1013, about 1012to about 1014, or about 1013to about 1015GC per mL). A composition containing an AAV containing a nucleic acid provided herein can be administered any suitable number of times, at any appropriate frequency, and for any appropriate duration. For example, a composition containing an AAV that contains a nucleic acid provided herein can be injected once, twice, three times, four times, or more than four times into the spinal column of a mammal (e.g., one to three times, or three to four times), at any appropriate position (e.g., along the rostral-caudal axis. For lumbar puncture, a composition containing an AAV that contains a nucleic acid provided herein can be injected in an amount of about 1 pL to about 50 pL (e.g., about 1 to about 5 pL, about 5 to about 10 pL, about 10 to about 25 pL, or about 25 to about 50 pL) can be injected at a titer of about 1011to about 1015GC per mL (e.g., about 1011to about 1013, about 1012to about 1014, or about 1013to about 1015GC per mL).

[0069] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

[0070] EXAMPLES

[0071] Example 1 - Identification of REs

[0072] To develop tools to treat pain by targeting specific neuron populations in the dorsal horn of the spinal cord using AAV, the cell populations most likely to influence the perception of pain were first identified, and SNAIL methodology was then used to identify and prioritize REs most likely to selectively target the underlying populations (FIG. 1). SNAIL methodology is described in detail in, for example, WO 2020 / 257520 (see, e.g., pages 4-6 and 21-26).

[0073] Identifying candidate cell populations: A database of cell types and molecular markers for the primate spinal cord was established as described elsewhere (Arokiaraj et al., supra). There was strong correspondence between the neuron subtypes of the rodent and primate, providing a solid foundation for translational research. Using mechanical allodynia in mice as a pain model, neural populations in the dorsal horn of the spinal cord were identified that underlie pain transmission (Peirs and Seal, Science, 354:578-584, 2016; Peirs et al., Neuron, 87:797-812, 2015; and Peirs et al., Neuron, 109:70-90.e7, 2021). As shown in FIG. 2A, there were several signals from light- touch mechanoreceptors (Touch), such as through GLUT2 and GLUT5, that can transmit to GLUT11 (Pain), and this pathway is normally inhibited by interneurons such as GABA2,3,4,5. Transgenic mice were developed that expressed chemogenetic molecules in select subsets of dorsal horn neurons. These molecules, called Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) (Roth, Neuron, 89:683-694, 2016), are cell-surface receptors that are activated by pharmacologic compounds that can be taken orally. By design, these drugs do not activate receptors elsewhere in the body, and only interact with the surface of the cells expressing DREADDs. Notably, there are DREADDS available that control neuronal firing, both activating and inhibiting subtypes. By inhibiting target neuron populations with DREADDs, mechanical allodynia in the mouse models was alleviated while leaving movement and normal sensation intact. From these analyses, the identities of multiple subsets of dorsal horn neurons that transmit or gate mechanical allodynia were confirmed (Peirs et al. 2015, supra, and Peirs et al. 2021, supra) demonstrating the efficacy of DREADDs to control neurons and identifying neuronal subtypes as targets to treat pain. Although successful, this technique relied on transgenic mice, limiting its translational potential.

[0074] In addition, candidate neuron populations were prioritized using human genomewide association studies. Single nucleus ATAC-Seq (snATAC-Seq) (Welch et al., Cell, 177:1873-1887. el7, 2019) was conducted in the Rhesus macaque dorsal horn to find cell population-specific REs that define these dorsal horn cell populations (FIG. 2B). Cell populations defined in macaque have equivalent species integrated (mice, macaque, and human) cell type nomenclature as shown in TABLE 1 and described elsewhere (see, e.g., Arokiaraj et al., supra). TABLE 1

[0075] Next, linkage-disequilibrium score regression (LDSC) (Finucane et al., Nat Genet, 47:1228-1235, 2015) was used to identify which sets of REs were associated with the predisposition to chronic pain disorders (Kupari et al., Nat Commun, 12:1501, 2021).

[0076] These studies revealed that several dorsal horn neuron populations (e.g., GLUT2 / 5 / 11 and GABA2 / 5) previously implicated in pain signaling were enriched for human markers of chronic pain at various body sites (FIG. 2C).

[0077] Machine learning prioritization of spinal cord REs: The snATAC-Seq dataset of the Rhesus macaque dorsal horn was used as input to prioritize candidate cell population- specific REs using the SNAIL system. The sequences with the strongest scores are set forth in SEQ ID NOS: 1-5 and 21-97:

[0078] CCTGAAATTTGGTGCAAGCAGCAGGCGCCAGACCCATGCTTGGTCCCA

[0079] TCCTCCCAGCATCTTTGTTTTGTCCTGGGTCGCTCTGCCAGGAGCTCCCTGGA

[0080] ACCTGGGAAGGAAGAGAAGCGTGCAGACAAGCGTCAGCACCCCCACCCTCC

[0081] ACTGGCCTTGGGGAGCAGGTGTCATTAGAGGAGGCTCAACTGAAGCCTGCAT

[0082] CTGAAACAGATTCTCATAACTCAGGCGCTGTGGTAGCCAGCTTCTCTCTACAG

[0083] TCAATTACGGCCAGCGGGGCCCACTGATTATTTTTATAGCCTTTCCTGGAGTCA

[0084] TGACAAGGAAAACTAAATGATCCACAGGACCCCTTTCATCACTAACAGCCGGG

[0085] TTTCACAGTTGGGGGTGGAGGAAGGAAAGAAAAATCACTTTTTTCCCCACTTT

[0086] GAGTTAATCAACAGAACTCAAGTCTGGGCAGCCAATATGTTGGTGTTGAGAAA

[0087] AGCTGTTCTCAAGCAGAACTAATTGGCAACA (SEQ ID NO: 1 ; R1_GABA3#3)

[0088] CCCCAGAAGCTAGCAAGAAGGAGGGAGAGATCAAGCAAAAGAGGACT

[0089] GGAAAGTGACCGTCGGACTCAGCAACATGTGTGCCAACAGAAGCTTAGCAGC

[0090] AGTGTTTGGGGAGGGTGATGGAAGCAGACAATTCCTTAGCTGTGAAAAAGAG

[0091] GAAGAGAAATACGACAGGAGCTAGAAAGAAATATGAGTCAAGAGAGACCTTT

[0092] TTTCGTTCAATGGGAAAGACTTGATGGTGTTTAAAACCTTCTGGGAGGGCTGC

[0093] ACTTAAGAGAAAGCAGTTGAGTACACAGGCAAGAGAAGGGGAAATCAGTAGT

[0094] GGAAGGTTCCATTGGGAGGGATTGCTCTGCAGGAGCAAGGAGAGAGTGATGC

[0095] AGGTGTGGGAAGGCTGGGAGGTGAGGAAAGTGGGGGATTTCATGTCTGTGCA

[0096] TCTGCAGGTCTTGTCCCTTCTACACCTGTGATCTTCACTGCCAGTCACAGACTG CCCACAGCCAACAGCCCCTCTTCCCTCACTCTTTA (SEQ ID N0:2;

[0097] Rl_GABA4#10)

[0098] TTGACCATTTGACACACAGCACTGCAATGGGTGATGGGTAGTTTTTTTT

[0099] TTTTTTTTTTCTTAAAGTGTAGCTTGTTAAATTCTTACTGAAAATAGACTCCAAA

[0100] CTACATTCTTGGCAGGGCACGAGAGAAAGGCCTGACTGTTATCTGCTGAAGCC

[0101] ATGAAACTGCTCTCAGGAAGCTGTTGGCTGCCAGGAGAGCCCCAGCTGAGGA

[0102] GAGCGGCTGAGAGAATTACCTCCTCGGGCCCACAGAGGGAGGGAGGCCTGCA

[0103] GGTCGAAGCCAGCCAATGCTCAGAGCAGCGTGGGCGGCCTGGAGGGTCCACA

[0104] CAGCACTTGGCTACCGAGTGGCAGCATCATTCCTAGCAGGCCATCTTCTCATTC CACTTGTCTGGCATCGGCAGGGCTGCACAGAAGCCCAAAGACTGAGCCAACT

[0105] GGGGTGGCAGAGAGGAGCAGTAAGAACAGAATGAAAAAAAAGAAAGAAGA

[0106] AAAAACTCCCTCTAGCTAAATGCTATCCTACC (SEQ ID N0:3; R1_GLUT11#14)

[0107] AAAAGTCCAAATCTCCTTGAATGGCACACATTTTATCTGAGTTGTGGTG

[0108] TGCTTTGTAAACATGGAAGATAAAATGACTTGGCTACCTGAATGCTTTGTTCCT

[0109] TGAGAAGAAATCATTCTCTTTCCCCCCTTTTGGCATATTGATTACTCATTAATCC

[0110] AGCAAAACAAAGGATTAATTAATCTCCATGGTTCAATGAAGAATAAATTAGCA

[0111] CAGTTTAAAATGGGCCTGATCCAATATTACAAATTGGAGGAACTGTTCCCCACT

[0112] CTCTCTTTCCATTCAGTTCTCCAAACTTCATCTCCAGAGAATGGCATCAGAAGG

[0113] CTAAATTTAGTTTGTGTGATATTTGGTATTCAGTCTCCTTCTTTAGAATTCAGTC

[0114] CCTTGAAGCTACTCATTGCTTTTCCCAAAACCCACTTTAACTATTTTGAAACAC

[0115] GTGAAGAGCAGCATTTTTTAAAGTGCTTTAGATATATCACTTATCTGAAACAAC

[0116] CCTTTTTTATCTAGAAAA (SEQ ID N0:4; R1_GLUT2#11)

[0117] TCTATTGGAAGAAACTGGAACTAATAATAATTTTGATTCATAGTACAAAT

[0118] AAATTTCTAGTGATCATAATTAAGAAACTATTCTAATTCTTACTTGACATAGCTA

[0119] ACTCTACAGAGATTTTGTTGATTGTGGCAAAAATAACTTCTTTTTTTTTTAATTT

[0120] CCTTAATTTCTGGCGGTTTCTATTTAGAGACTGGTACTGTTAAAAACTGCCACT

[0121] AAGGCTCTATTATGGTTCTGCTGGGGGTTTAATTTATTAGATTTGGACCTAACCT

[0122] TATCAAATGAGCTTCTAACTAATGATTATACTCTAAAGACCACTTTTACACTAAT

[0123] TGGCTTTCCCTGTTTAATTGTTTTACAAAATTGATAATTTAGTAGAAATATAATA

[0124] AGCCTTGAATATTTTAGATAATTTGGCCCCTTTGTGGTCTTTAGAGGGTTCTCAC

[0125] TTAATAGAAAATTGGAATACACATTGTAAAATTTAGGTCTTGCTCTCAGCACAT

[0126] GATTTATGAAGGA (SEQ ID N0:5; R1_GLUT2#3)

[0127] GGTTGGAACAATCAACACGTCCAGCATGAGATATGGGCCATTCGCCGA

[0128] CCCAAGTCTAGGCTGTCTCAAGGCCCCCAGGCCTGGTGCCATGGGCACCCCAC

[0129] CAGTCCCCAGCCTGAATGTCCACGCAGGGGGTGGAGTGGGCGGGAGGGCTTC

[0130] CAGAGTGTGCTGGCACGGAGCGGTCCTCACTGCCCCTGCCTGAGCCTCATCA

[0131] GTGGCAGACATAAACTAACTCGGTTCCTGGGGACCCAGAGGAGCAGCAGGAA

[0132] CTGTTTGGTAGAGAATTATAGCTGTGGGTGAAATTAGATGTACTCTGAGCCCCA

[0133] AGGCTGAGAGAAATAAAAAGCATTTTAAATGGAGTGTAGAATTCATATTCCTC AGTCACCTTCCAATAAATAAATCTTGTACCATTTTTTTCTGTAAACAGTGGTTAC

[0134] ACTCAGCTTCCATTGTCTCCACATCCCATATGCCAATTGGCAGGGAGTAGCAGG

[0135] GCCGTGCTGGACTGGGAAGGAAAAACAT (SEQ ID N0:21; R1_GLUT11#7)

[0136] ATGTAACTCAGCTGAAGAATCATTATAGGATTTTTTTAAAAACTGGAA

[0137] ACTTCCCCCCTGCCCCCAGCTGTCTACAGAAATAGATTATTTCTTCTAATTTCC

[0138] TGATTTAGTTATAGTCATAGTGTACATAAAAACAGATTTGATTTTCTTGGAAT

[0139] CAAACATAAAATGAAAGCCAAGGCCTTATCATTGAAACAAGACAGCAGTTTT

[0140] GCAAAGCTGTGTACATCCTTTGAACTCAGATGACATTGTCTCAGAGGAAAAG

[0141] ATGTTTGATGAGTTTTTTACACTTTAAAGCTCTTCCTAAGGAAAATGAGCTGG

[0142] GAATCTCCTTTGGTAGAATTGAAGGGGGGTGGATCATTTCGTCTTCAAAAGAT

[0143] GAGGGGAAAAGTGTCATTTTCTATTTCCCTTAAGTATCATTCTTTCAAATACT

[0144] TGTGTGACAAGAGGTTCAACAGGCTTAGATTTGGGTTTTTAGTTTTCCAGAAA

[0145] TGCATGGAGGACTTAAATGGCCC (SEQ ID NO:22; R1_GLUT5#92)

[0146] ACGCGTAGAACACGTGGATCCGTTAGAGGATGCCTCTTGGTTGCTTAA

[0147] AAAAAGAGTCAGTCCATTTAAAATTGTTAGTTTTCTATAGTCTTTCAGAACAT

[0148] AGAATTCATTTAGAATTTCCAGCAAAAGTACAGAGAGTCTTCAGGGAAGTTG

[0149] AAGAAGATAAAATAATTCTGAACACACTGTACACTTTCAGAAGAAAAATATT

[0150] AAGAGTAAAAATAGTTTCCCAACTCAGTCATTCTTTGGAACTGATAATTTATT

[0151] CCTGATGGTTCCCCAAAAGAGGGGAAGAGAAGCAAAATAATATATTTGGCTC

[0152] CTCTAATAGATATTATTCATAGCTCTTTTCCCTCATAGGATAATAGACTATTA

[0153] GAGCTTCTATAAAATCCCTTTCAGTAATTTTATGGTGGGTCTGCATTTTGTAA

[0154] GCCATGTTTTATAACTGAGAAAAGAAAAAAAAAAAACACAACCACCAAGAG

[0155] AATAATTTTTACAATAATTCATTCCACTGATTTTATTAT (SEQ ID NO:23;

[0156] R1_GLUT6#97)

[0157] CATTGTAGGGGGCAAGAAACAGCTGGGTTTTGTCCAAAACGAATGGT

[0158] GATGGTTTCTATTGCATAAAATACGATTGTATATGTATAAAGCTCAACACCCT

[0159] ACTTACCTGGATTATTCTGGCAAAAGTTCTATTTTAGTCCAAGTTACCTGCTTT

[0160] GGGAAGAAGGTTGCTTTTCTCAGGGGGTAATTACCAGTGCAATCACAGCACC

[0161] ATCTGCTCGCCTTATGTTCCGTAGATGACTCGAAGGAGATAGTTTCAGGAGA

[0162] GACCTGAAAAAGGAACATTTCAGGAGGGAGGAAATGAAGATAACCTAAAAG AAAGTCATAATTAGGAACTTAAAGAGGAAAAACATATGGTATTGAACATATA

[0163] ACTTAGAAAGCAGTGTCCTAGACCCCTGGCCCTACTGAGGTTTTCCCAGGAGT

[0164] AAGAACTGGGAGAAAGTCAAGGCTAGAACAGAGTGGCCAATTAACCTAAAG

[0165] GGAATCTCCAGAGGGGCTCAGCAGAGAGAAATAGC (SEQ ID NO:24; GABA1)

[0166] CCTTCACCTGCTGTTTGGTTGTTTTTGTGCCACGGGTCCTGGGCTCCTT

[0167] CCTCTGTCTGGCCTCGGAGCTGGGCCTCGCTCACATCAGCACCATTTTGGCTG

[0168] TAAGCTGCTGGAGCAGCTGCTGCCCGGGAGGGCTCTGCTCATGCCCTGAGCT

[0169] GACACATGACAACTTCTATTAGGCTGAGGTGCTCTGAGGGAAGCCAGTGGGC

[0170] CCTACCAGCCATCTCCCTTCTGTTTCCCCATCTGCTGGCTCCCAGGGAGGTGG

[0171] TGGCAGAGCTTAGGCAAAAGAGCACATCCAAAGTGAGGACATGGTGAAGAC

[0172] TTGGAGGGATTGCACTCTCCCTCTTTGTCAGATGTTGCCTGGCAGGGGTGGAA

[0173] CCTGTCCCTTAGGAGAAAATGGGGTGACTCAGGCTAGCCTGGCTCATGCTGA

[0174] GGACGTCCAGTCTGCCTCCCACCATCCCTGCTGGGAGTCCCCACATGGAAAT

[0175] CATATTTGCAGCTTGGGTTAAACATGACCAGGT (SEQ ID NO:25; GABA1)

[0176] GACCTCCAAAAATCTTTTATACCCTGGCCTTGGTCTAGGAAGAATTCA

[0177] AGGATCCACAAGGTGCATGTAGAAATTTGTTGGAAATCAGACAGCCACAAAC

[0178] ATAATTGGTAAATGGGATGGAAGAGGAGAAAGCTAAAGACTCGACATAATTT

[0179] GGCTTAGAAAAACAAAGACCAAGTGGTGACTCACTAACAATCTTCAAGCACG

[0180] AGAGGGGAATTTAGGGGAAGACGATGAACAGCTGTTTCTGCTTCTACTGGGG

[0181] CTTGGCTAATATAACCGGAGTTTCCAGAACAGCAGGAAGGGCTGAAGTAGAA

[0182] AAAGAACTCCAGTTCCTGACAGCTGTGAAAGATAGTCATTGATTGCCATGGG

[0183] GGTTCCTATCTTTCAGTTCTTTAAAATCGGATAAATGATAGCTTCTTAGAACG

[0184] TTCAAAGACCCCCTCTCCCTGTCAGTGTTTCTCAAAGTGTAGGATGCAGGCCA

[0185] CAGCTGGTAGAGAAGACATTTGGGTGCAGAACAG (SEQ ID NO:26; GABA1)

[0186] ATCCACCTATGGAATGAGATTTTTTAAAGCAAAGAAAGTGTCTTCCTA

[0187] GGAATGGGCTGCATGATGCCAGTCTCCTGCCCTCTGGCCAGTCTCACAGATG

[0188] GGCCACTCTGGCCAGCAGCCACCCAGCACTGCCCTTGAGCATTCACAGCCTA

[0189] CTCAGTCCTGGCAGGCACCCCCTGCCAGAGGCCCCTGGGAGTCATCATGTTG

[0190] TAGGCCTTTACAGTAGAACATTGCTCTGCTGCAGGGGCAGCTGGCTGGCCCC

[0191] AGGAGATCACCGGCAGATGTTACACCTCCCTCCCCACAGTCACCAAGAGCAA GGGGAGGGAAAAGGCAAGGGAGTGTGTCTCCTATGGGATAGATGGTGGCCA

[0192] AGCAGCAGGCCAACGGGTGACCAGAGAGGCAATGCCTTTAACAAAAAAAAT

[0193] AAATAAATAAAAAATAAAAACAAAAAACCCACTTCTTGCACAGTGGTAGGTC

[0194] AGAAAAGAATATACTTGTTTTTTTGCTTGGTGTGGGTT (SEQ ID NO: 27;

[0195] GABA1)

[0196] TCTGTCCCCAGGCCCGATGACTCGGCCTCCATCCTCCCTCCCCACCACC

[0197] CCAATCTAAACCAGCCCAAAGGCCAGGCAAGAGGACCCCTGCTCCTTGCCCT

[0198] TCAAGGAAAAGGAGTCCTCAGAGGCCAGCCCAGCTGTACACACTGCACCCCT

[0199] GCTTGTGTTCCCTGAAAGCAACTGTCCTTTTGTTAATTCTGCCATTTCCCAAAG

[0200] TACCAGGAGCATCTCATCTGACAGCCACAGCATCTGTTCTTCTTACTCTCCAG

[0201] AAGGTTGGGTGGGAGCCGTGCCAGCTTTTCCTCAAGGCTCTCCCAGCCTCAC

[0202] GCTCCTTCCACCTGAGTTGGACACCCTGCTCACCTGGTCCCCTAGGGGCACCT

[0203] CCAGTTCCCTCTCTGCATCCCTGGCTCCTGGCACCCCTTCCTAGGACCACGGT

[0204] TGGCAGCCTCGATAACCATCTCCCCTCCCGGGGTCAAGCTCACCTGGCCAAG

[0205] CAGACGGACACCCATCCTGTACCTTAGGAG (SEQ ID NO:28; GABA1)

[0206] TGGCCTCAGGCCCAGCCCTACTCTATAACCACTGCTGCTGTCAATATC

[0207] CAGCTAAGGCTGGTCTGAGTTCTCTTTACTGTCAGTTTTTCTAGGCTTCTGAA

[0208] GTTGACAGTGGCATTGCTCCTTCCAGGCCTGGTGGGTACACGTGCCAAATCAC

[0209] AGCTTTGCATGGTTGGTAAGGTATCCCTTGGCACTCAGGCAGGCTGAAGTTGC

[0210] CTCTCATCCCAGGGACTCCTGCACCACAGGGGTTGTCAGGGAGAATGCCTGA

[0211] AACAGCTGTCACCCCTCTGCTTGCCAGACTGATGTGATTGGCTTTGTTGGTAT

[0212] TCTGTCCTTCCCCACACCTCCTAAAGTTGTCCAGTTCGTAAGCGTCAACTTTC

[0213] CAGAGAAAGAGGACTGTTCTTGCTTTGTCCTCCTCACCAGGTTCATCTTTTAT

[0214] TCTGCATCTAGAAGGGATCTAAAATGTGATCATGTCACTCCCTGGCTTAACTT

[0215] TTCAAGGGTTTTCCCAGCCTGATGCTCAG (SEQ ID NO:29; GABA1)

[0216] GGCCTATAATTTCTAAGAAGCCAGGATACATTTCTCTGACAACAGGCA

[0217] ATTACTTTTGTGCGTGTGTTGAAGGTGACATGATCTTCTTTAATTTAAAGGCA

[0218] CGGGAAGACTGCTTTATGGAAAGGTGAATCACCTTTCCCTTTCCTCCTCCTCG

[0219] CGACTTGACTTGTACCCCAGATTTGCTTCGCTAATTCATTCCTTGCTTGTCTGT

[0220] CATGGAAAATGGTTTGTTACGTTAATATCATTCTTATGTTTTTCAGTTATGAAT AGCTTTTCAAGGCTTAATGTAGCTTTTGAAAATTCTCTTATTACTGATCTTGGT

[0221] ATTATCACCGTAGGAGTTAAAGTCATGAGACAATTTTATGGGGGTAATTTCTT

[0222] AGTAAAGTAAAATTTTGTGCTGAAATCCTGCTGGGTTCTACACATGTTGATGG

[0223] ATGACCCACTGTTTAATAGGACATTTTACTGTTCATATATGTGAGGAATGGGT

[0224] GTGCTAATTAGAGACTTAGAAGACA (SEQ ID NO: 30; GAB A3)

[0225] GTCTAAGGCACATCTTCTCCAAATGAGAATTGTATTCATTTTTATTAGT

[0226] AGTAGGATAGTTAAATCACCTTCATAGAATAACTTACGGGTTTTTAAAACAA

[0227] ATCTGAAAATTTATTTGAAAGTTATTAATGCTAATTTTATTATTAAAATTGTA

[0228] ATAGTCTTACTCTAAAACATTGATGATATGGTGCATGACCAATATTATCATAG

[0229] CTGCAAGATTAATTCTACAGTCACTGTATTAAACATTGGCAACAGTACTAAAT

[0230] AGACACACTCAAGTGGCACATTAATGCTATTGCTAACATTTTTGTCTATAATA

[0231] GAATTTGAATAGGAAGAAAAATTTAAATTACATGTTTTCTAAAAGAAGGTAG

[0232] AAGCAGGGATTGTAAATTGGATAAATATAAAAAATCCTTCAACTACTGAAAA

[0233] TGGTTAGCAGTGTACAAAATGATTTTGACAAGCGTGGTGTTTAAAATGGTAA

[0234] CTTCTGTAGACTCACACCTATAAGGAAATGT (SEQ ID NO:31; GABA3)

[0235] GGTGAAAGAAGCCACCTGGATCCCAGCCAGGGAACTTGGCCCAAGGT

[0236] GACTTGGGCTCTGGCCGTCTAGTGGGGAGGCAAGGCTGTTACAAATGGCTCA

[0237] CAGGAGGGTAACGCGGCACAGATTGAATTTATAAAATACAAGGCAACCACTG

[0238] GAGCTAAAATTTTATTGTTTATAAAATATTAACCTCCTTATTACTTTTCATTGC

[0239] ACTTTCAAATAAAGCACAGCAAATTAGAAGCAGCCTCTGAAACAAATCCATG

[0240] GGTGATTAAATCAGGGAATGAATTAGAAGGGGAAAGTTGAGAAAGGGGAAG

[0241] ATGGAGACTGGGGTGGGTGGGGGGAGGGAAGAGGCAGGCGTGGGGAGGCA

[0242] GCCTGGGGCGGAAATGGGAGAGGGGCCCTTCCTGGCCTGCCCCGGGAGAGA

[0243] AGGCTGTAGCAACTCCCTGAGGAGTGGAAAGGCGAGTGGTGGGAAGTGCAG

[0244] GAGAGAGGGAACTCTGCAGGAGGGAGCTCTGGGGAGTGCA (SEQ ID NO: 32;

[0245] GAB A3)

[0246] TCAGGATTATTGTGGTTAGAGGAGGAAAGTTAGGATGCAAGACAATTT

[0247] TTGAGATCCACAAATAAACATGAGAACAGTGAAAAAGCAGTTAATGCTATTT

[0248] AAGGGATGAAAAGAATAGGCAGGATCTGTATAAATATATCAGGAGTAAAAC

[0249] ACCTAGAAAGCAAGTAGGCATGAAATTAATAATTATTTTTAAATGCTAAGGT TTTGAATTCATTTTGCAACCTGACTTTAATAAGAAAAGTAGGCAAGAGCATTT

[0250] TGCACAATTAGGTAGTAATGGAGAATTTGTAAAATACCTACTAATAAAATGT

[0251] AGCAAAGCGAATATTTTTCAAAATGAGCATTGAAAATCATTAGGTTTAGATA

[0252] ACATTATTCTAATATGATGGGAAAATTATCTCACTAATTTGACAAAAAGAATT

[0253] TATTTTGAAATAGCAAAGTAAGGTAGAACAAGTAATACAATTAGGGAAAGTA

[0254] CAGGCAGTTCAGTTCTTTCATCAAATGTAGTTGGC (SEQ ID NO:33; GABA3)

[0255] GTGCCCAGGAGTGGGTGTTGCAGGGACTTTTGGTAAACTGTTAAGAAA

[0256] TAAGACATTCAGCTACGTTTATTGTTAAAATATTTTAAAGGACTTTGTGCTTT

[0257] GAATGAGATTTTTGCATACGAATGTTTTAAAACCCTCTATGGTGAAGCATCAA

[0258] ATGGAAACTGAAATGATCAAAAATGTATTTACCCCCTTCTGTGATCTTATTAT

[0259] TTCATGTAAACCACTGAGATTTCACGCTGAAATAATTATCATTAGTCCAAGGC

[0260] GCACACACAAGAACAAGGGAAAAAGCTACTTTCAGGGTATGTTATCTCACCT

[0261] TTGCGATCCTTTACATTTGATTGTATGCACGCATTGTAAGAGGAACTACTTCA

[0262] ATTAACTATGAGAACAATATTGATTTGCATTCTGATAATGATCACAGGACATA

[0263] TTTAGAAATAGCTGGGGGTAAATATTTCTATTAAATTAATAGTGGCAAAATTT

[0264] TATTTAATTAAACTATAAACGAAACTGGT (SEQ ID NO: 34; GAB A3)

[0265] AGATGAGGAAAGAAAAGATCAGATGATAGGAAACGTGTGTGATGTCA

[0266] ACTAGCTGCAAGTTGCTAGCAAAGAAAGTCCTATACTTCTTGAAAATGCTGT

[0267] GGGAACAGTTCCTGCTCCCAGTAGTCCTGCCCTATCTAAAGAAGGAATTCGT

[0268] GTCTCTAAACCTACTATCATTTCTCTTCCAGATCTCACCCTCCCCAGGTGGTG

[0269] AGGCCCCCTCTTGCCCCAGCCCCCTCCCTAGCAATGACACCGCTGTTTCCTAC

[0270] AGCAAGTGTAAAGCACACACAGACTCGGTTTAGGGACAGTTAATTTCCAATT

[0271] CTTTCCAGATGGGTTGGAATAATATCCCCAGGTGGTAAATACATTATTAAGA

[0272] AGGCAGTTTGGTGTGAAATTGGCTATTAAAATATAGATAAAGTTTAATATATT

[0273] GAGCTCCTTGTCAGAAAAATCACTTTAGAGGGAGCTTATTATTTGTTTGATAT

[0274] TAAAAGAGTCACAGCTCCGCTCCAAGTGACAAA (SEQ ID NO:35; GABA3)

[0275] TCAGTGGCGCCTGCGTGGAGGGGGCCCCCTAATCGTGGTGCCTCAGCT

[0276] TTGGAATGTTCTGGGAGGTAGTCAGAACAGGCATGGCTGCCACTGCTGACCC

[0277] TCCCACCTGGCGAACTCTTTCCGGCTAATCGTTCTGTTTGTTTTCCCAGAGCA

[0278] GGTGGGGGCAGGGCCTCTTCCATCCATTATCGGCTCCCATGGCGCCAGGGAG GCCCCGTGTGCACACGGGCTCAACTTTCTGGTTTTGCTTGAATTAGCAGGAAG

[0279] CAGCAGGGAGCGGGGTTTGAGATATCTGAATATTTGTGGCTGTGCAAACCCA

[0280] GAATCAACAAATAATAATCAGCAAATCAGGAAAAATTACATCAACAATCAAC

[0281] AAATAATAATACACCAGGAATCCATTGGTTATGAGCACGGCTCAGCACGGCG

[0282] ACATTTCTCAGCTCACTGATGTCCCTTCCTTGGTGAATGCGGAGTCCTTCCTGT

[0283] GTGCCGAGCCTGGCGCAGGGTGCTGGGGCCCA (SEQ ID NO:36; GABA3)

[0284] TTCCACTGAACTCATCAATAAGAAATGAATTTTGGTAACTTTCAAGTC

[0285] CACTATATTATTTTTCCTTTTATTTGTAGTGAATTTTTCTTCTCTCAATGGTATT

[0286] ACTGCAATAAGTCAGTCATTATTTCTTTATGTTTTCCTTTCTTTCTTTAGTATTT

[0287] CATTTCCCTTTACAGCACATCTTGTGATCACAAAGAAAACAGAGATACTTATT

[0288] TCTTTTAATGGTTGAAATTGCTGCAAGTAATTTAGCTGTCATTTAATTTGATTC

[0289] CTCTCTTCTGTCTGTGGAGAAATATGTTTAATGTACTGGAAATTATGTTAATA

[0290] AGAAAAAGCATTATCTCATTTAATGAAATAAATATACAAATATTTCAAAATA

[0291] AAATAAAGAGGCAATCTTCACTGAGAAAAATGTCTCTTACGGTTTGCATAAA

[0292] ATAAACAAATGAGAAAAATTTATGAATACATTGAGACACTGACAAAAATTAC

[0293] ACCTACATTCGGGGGTTGGCATGGCC (SEQ ID NO:37; GABA3)

[0294] AACTTAGGGTCAACATTTAAAGAGAAGCCATTTCCTATTTGTTTTGAA

[0295] GCTGCAGAAACCCCCTCCCCCTCCCCTGTTCCTTCTATCTATTTTGTTCTTCTA

[0296] ATAGCAGGAAAGGGCAGAATTTTAATAATCTCTCTCTAGAATGACTTTCTTCA

[0297] TGCATGAATTATTTCTCATTTACAAAGGTTTGGGAGATGCTTTTTCCCCCCTTC

[0298] CCTTCTCGCTGTGGAAAATGATGACTGTTTTCATTGCTAGACACATTACTGTC

[0299] AGAGCTCATGAAGCTGATGGCCCAGCCCATTTTCCCACGGAGCCCCTCTCCA

[0300] GCAGCAGGCTCAGCTCGTCACGTCCCTGAAATAGCAGCTCTTGTCAAGGTAT

[0301] GCACGAGAGTGTCGCAGTCACGGTGGCTAATGGCAGGTGACTGCTGTTGTGA

[0302] CTCTGCTGGCGTAGAACAAAGGCGGATGAAAGGCCGTGGAAGGCCAGCTCC

[0303] CCTCCATTCATCCTCACGATAACCTCTTGGA (SEQ ID NO:38; GABA3)

[0304] AATACTTGGAAAATGCATTTCCTGATGATAGAGAAATTGTGATTACTT

[0305] AGAAAATAGGGGCTGCAGAGGTTATCTTTAAAGAATCAACCAATTATCTAGA

[0306] CCGGTAAGTAAAGAATTGAATAATTGAAAGAAGCATATAATTGATTAAAATG

[0307] GCTGAGTTTCATTTTTGTCACGTTTATAATGTTCAATCAAGATTATCAAGAAT ATTTTGCTCCTTAATAGGATTACTAAGACTCAGGAGGGTGAAAAAGAACATT

[0308] TAGTATGAATCCGAAGTTGCTATGGTGTATAATGAGAGTACATTGTGTAGGT

[0309] GTGTATAAATGTATTTCGGGTCCCATTAGGTTTTCTATCCAGTGGTTCTCGTAT

[0310] CTCCATCTTGAGAAATTTAAAAACTGATTGACAATTACCCACCTCCAGTGGTG

[0311] GGAAGGTGGAGTGCTTACTCTGACAACCTGGACTGGACTGCTAATTATATGC

[0312] CAATTGTTGAATGGTGAACTTCTTGAGGCTCT (SEQ ID NO: 39; GAB A3)

[0313] ATATGAAGCTCTAAATGGTTTCAAAACACTTTTCTTGGGCCCCTAATAT

[0314] TGATGTAATGCATCAATTGAGCAATCAGGGTGTTATTCTTTAAGTGATAAAAA

[0315] GCACGCAACTGCTAATAAAAATATACATAGTTCAGCCCAGATGACATTTTGA

[0316] TGATCAACCAGTACATAAATAAGACTGTTTGCTTCTTTAAAGAAGTGGAAAA

[0317] AGAGCAGTGATGCTTGTGAAAATCTCATCCACTACATTTACAAAACAATTTTC

[0318] CAAACATTCCCATAAGAAAATATGATCCTTCTCATTTCACAGATGGGTACACT

[0319] GACACTGGGAGGTGAAGCGATTTTTCCATGGTGAAACAAAGTCAATGAACCA

[0320] TTTGGAGTTTGTTCTGAAAGTTCAACATTTTTTATTCTTATCTTTCCACAAAGG

[0321] CTTTATCCCTGTTTGTTTAAATGTAAATTAGACAACAATGAAAACAAATGAGT

[0322] CATCATTTAAAACTTTAAATTGCAAATTT (SEQ ID NO:40; GABA3)

[0323] TCAGTTGGTAAGCAAATTAGATAGCAATCAAGGTTACACAAAAATTAC

[0324] ACTGTCTTAAGAATAAATGTTAGCCGGGACGGCATAAATGAACTGATAATCA

[0325] GGTAGGAAGAGGTTATGGCAAGGAATATGTTTCCTTAAACAACTTTTAAAAC

[0326] TAACAGAATTTTAATAAGGTATTACCCCCAGTCAATAAAATTCTACTGAAAA

[0327] ATAATGAAACCTGCAAGGGTGATGGCTGCAATTCTGCAGGCACCCACAGAGA

[0328] CTCTGCAGGAGGCCTGCACGTGCTTCTTCCCTGCCTCTCCTACTATTTACAAA

[0329] CATAATGAAAATGAAAATGAATATGCTGCTTGACTGCTACTCTTAACAGCAA

[0330] GGACTGTGAGAAGAATATTGTTTTAGTTTGTCTGTCAGGGTGACCTTTATGAA

[0331] TTTCACTGTTGCTAGAAAACTCAGGCTGATTGGACAATGTTCTCATTTCTTTA

[0332] AGAAAGATTTGAAAGTTCACAGGAACTACTTTT (SEQ ID NO:41; GABA3)

[0333] CATGCTTCCTCCTCCGTGCCCTCCTTCCATGCCCTCCAAGGAGAGCACG

[0334] TAGGGCCGTGGAAGGGGCCTCGGCAAGTAAGGGACCTGCTGCTTAGGCTGCA

[0335] TCAGCAGATGGCATCTGCTTACAAAGCCATCCTTGGCTGTCATGACCCTTTCT

[0336] GAAAGAGAAGACGGATGAGGGATGTTGTTTTTCTTTCCTGAGCCTATAATAC AGCAACAGAGATGAGATGTGTAAATGCCTAGCCTAGAGTCTGTCTCATGGTA

[0337] ACTGTTCAGTGACTGGCAGCTACTATGGTTATTAGTAAATTACCATCTGCTTT

[0338] AATCATTCTCCTGTGAGTTTCATCTGAACCTATTAGTTTCCCAGAGCCAGCTT

[0339] GCCCTGGGGAAAAGTCACAAAGCTAAGAACAACTTTAATGTTGGCCTCCAGA

[0340] CTCAGTCAGATACCTGTTTGCTAGAACACTTTGAACCAGGGTGTGGGTCCTGC

[0341] TCCATGAACATGGTGTGAATTACCCTGTAAA (SEQ ID NO:42; GABA5)

[0342] CCGGGAGTAGTGGACTTATCCAGGTTCCCAGGCTCAGGGAAGAGGCTT

[0343] ACGACAGCCCAGCCTCACACCTGGGCAAGTGGCCCCCATCAACCTGCCCAGC

[0344] TGCCCTGTGTCTGATTTGCCCCTGGCAGCAACCTAGTTCCCATGCAATTGGCC

[0345] AGATAGTTTTCCCCAAAAGCAAGAACCAACCCATCCTTCCACTGTGAAATGC

[0346] CTCCCACCCTTGCTTGGGTGACTCGGTGCCAGGGTAGGCTGATATGCCAAAA

[0347] ACTGAAGCCTAAAGTGTCGCAGAGAACCACAAACGCCATCTGTCAACAGGG

[0348] AGTTAAGGCACGCTAATCTAAACACATCCCTGGAGATAGCTCTCTACAGCTC

[0349] ATAAATATATACACACAAAACCCTCTCCCAGAGGCAACAAAGCAGGACTGGC

[0350] TTGAGGCCACGACCAGGCCCGCCTCAGACACCCTGCCTCTCTCTCCTTCCTCT

[0351] TACCAGGGACTTGGGCACCAGGGACCCTGTCCATT (SEQ ID NO:43; GABA5)

[0352] AAAGTCTCTAAAAGAAAATAGATTTCCACATATATTGTTGGAATAATT

[0353] CTGAAATGTTTTGTTTATTCTTGAATAGGTGACACTGTTAATGCATTTTAGTG

[0354] GAATGAAATGAAAATTTAAATGGGGTTATCAGAGGCTAAATGCTGTCACTCA

[0355] GTGTCAGTGGTGATAGTTTGGGCATTGTTCCAGCAGATCATAATCTGAGAGAT

[0356] AATGACTCTAGTTCTGTTTTTCCCAGCTGTTCTTACTAGTGGGAACAGCTGCC

[0357] ATTGTTACACTTGTAAACTCTATGCTTGGCAGAACTCTTGATGGAGTCTGAAA

[0358] ATGCATTTTGTTTGTGACTTTCTTTTCCTGCATGGTAAAGTTACATTCTGATTT

[0359] AAAACAGGAGCAACACAGCATTGTCTGGTTGACACTAACGGTGCCTAGGAGA

[0360] AATTGGAGGAATTCAATGGCACTGAGCATGCATTACCTTGCATTTTCCCTTTA

[0361] TCCTGGTTGAATGAATCATTTGTGATATG (SEQ ID NO:44; GABA5)

[0362] TCTCGCACTCCTTGTTTAAGGGCTTTTCAGACTACACTGGTATTTCCCA

[0363] TTCTCTTTCTTTTCCCAAAATGGTTAAAATTCAAAATGGAAAACAGCTATCAC

[0364] ACATGGGCAATGACTAATTTTCCTAAGGAATTTTAGTATTCGGAATGTGTCTG

[0365] CTTGTGCAGGCCTGTGCGATAAAGGCTCCCTGAGTATGCATTGACTTGATACC CAGTAATGGCTGCTAAGGAGCCGTTATGTAGTGCCGTGTCAATGATCCGTGC

[0366] GCTGAGGACTCCCCGAGTACGCATTGACACGATAGACCGGAACGGCTCAGAA

[0367] GAAGCTGTTATGGTCTATCATGCCAATGATGTGCGGATATTGGCAGGGCCTCT

[0368] GGCCCTTCTGTGGAAACTATGGAGTCTCTGAAGGAGGCGGGGTGGGCGCCAT

[0369] GTGGGCGCCATCAGTTGAGCCTAGCAGCAGTTCCATCTTCCCTTTGCCTTGGG

[0370] TTTACTTCAGAAGAGTCAGCTCGGAAAAGT (SEQ ID NO:45; GABA5)

[0371] ACCTCTCAGAACTGGTCACTCAGCAAAAGCCAGATGGCATTAGTAAAA

[0372] GATGTTGGGCAAGGGTGGGAGGTTGGGCCAGTGATCTCTTAGGTTACTTTTGC

[0373] TGCTGAGTTAATAATTGTCACATTATTAAACAGCCCTTGGCCCTGGGAAGCCC

[0374] ACCTTCTGTGAATACAAGTCTCAGAGTAGACAATAGGGCACAACCAGCCATG

[0375] TTACATGACAGACTGCCAAGTGCATTCTTCTTACAACTCACTGGGATGGAGCA

[0376] CTCCTCAGAGAAAAACAAGCCCAGCAACATTTGTATTCTGAACAGGCATCTG

[0377] TCTATGTTTGTGACCTTTATGTTGTGATGTGTGTGAGCTGCTGTCTGGAATTGG

[0378] AAAGAATGAGGAAATATGTCACCTATGGCCCCTCTTGTGAAGTGTTAAATGC

[0379] CTCTGAGGCACTTCAGAAGGCAACAGAGCGGAGAGGATGAGTGATTAGAAG

[0380] CACATTTGGGATACATTTTTAGATCTAATTAG (SEQ ID NO:46; GABA5)

[0381] TCAACTTCCAGGTGAATAACCGGGTTGGTAATAGAGCAGGTGAGAACT

[0382] GAGAATGCTGGTCCAATCTGGATGTGCGTAGTTCAGTTTTTAGACCAAAGAA

[0383] TATCCTTACACTAATTCCTGTTTACACATGGCACATGGGTTTCATTTCACAGA

[0384] CCAACTCTGATCAGTGACAGCTGCCTGGAGCACCACAGGGAGAATGACTCTG

[0385] TGGCTTTGTCCAAGCTTATGGGGATAGAATGACATAGCTGATTAGCGATGTCT

[0386] GCCAAGGGGGTACAATAGAAGAGTTAAAGAATGTACGCTGTTGGTCGGGCAC

[0387] AGTGGCTCACGCCTGTAATCCCAGCGCTTTGGGAGGCCGAGGCGGGCAGATT

[0388] ATGAGGTCAGGAGATAGAGACCATTCTGGCTAACACGGTGAAACCCCGTCTC

[0389] TACTAAAAATACAAAAAAGTAGCCAGGCATGGTGATGGGCACCTGTAGTCCC

[0390] AGCTACTCAGGAGGCTGAGGCAGAAGAATGGTGT (SEQ ID NO:47; GABA5)

[0391] ATGAAAGACTCAGACTCTTCTCTGGGATCTTAATTTTTATGTGGCTTCT

[0392] TATTACTTTTTTATCCCTTCTAAAACTCTAAATCCCTTCAACATAGCTTTGGCC

[0393] CCTTCCACAGCTGAACTGGCAGCAGGCTGATCTTTTTATTGGTGGGTAATGGA

[0394] TTTAGTTTGATTGCTGACACGGGAACAATGAGAGGATAAACAATTCCATTTGT GGCCATGAAATTCCCCATCTGTTGAGCGCGTTCAGGACGGCAGAGCACTGGG

[0395] CAGAGACAGATGAATGAGCCCGCCACTCCTGATGTTAATGACATCCGTCATA

[0396] CAAGCTGAACATGTGGCTTCATAAATCAACAGCTAATAGCAACTCTGGTTGC

[0397] TCATGTCATTAATAAGCCATCCGTTTAAATACATAAGAGTTTGCCATAACATT

[0398] CAGCCTAGAGACCTGACTTGCAATTCTGATCTCACCAAAAAACCAGAAGTAG

[0399] TAAAGGATGCCGAAACAGCATTCTTCTAAG (SEQ ID NO:48; GABA5)

[0400] ACTTAATAAGGGGTTGCTGTTTTCTATCCAGATATAAATGCAATTACA

[0401] ATAAAACCATGTTTGTTTCCACTAGAGTTAGATGTTTAAACAAGTTGACACAT

[0402] TTTTGAAAACGCAGTAAGTAAAGATGCTTTGTTTATTTTTATTTCCATATTAAC

[0403] TTTAGTACTGACAGAGGATGACAAAGAGATAGCCTTGGAGACCACTGAAACC

[0404] ATTTCATTCACAAAATGGGTATTTTGTGAGTAAATGCACTGCAGATCGTTCCT

[0405] CTGTTGGGAACATTGCTTAGGGCTCAGCAGATGGCCACTGGAGTTTGAAAAC

[0406] ATCTGTTTGCTTTCTCTCCTTTTCTCGGTCCAGAATCCACAAATGATAAAGGCT

[0407] TTTACATAATAATGCTCTCTGCCTACTAGGACTAGGTTACAGCTGGCAGGAAC

[0408] AAGGAAGCGAGATAGCTTTGATAAATGTGATATAATACTGTCATGTTGTAAA

[0409] AACACTTTGACTCTGCGATAGACACTACT (SEQ ID NO:49; GABA5)

[0410] AGGTGGAAGGCCTCGGACACAGTGCTAGGGCTGGCCCACCGGCCTCC

[0411] ATCCCTCTACTCAACTGCTTCCTGCTAAATCTCACTCCCTCTCCTGCTCCAGGC

[0412] CTCAACCTGCTGCTCTGAGGATCAGCTTGGAACGTCTGCTCCGGAAGGAGGA

[0413] CCCTCGGGTTTCTTCACTGCCTGTCAGCAGCCCCTCTGCCTCCTCCCTGCCCCC

[0414] GCCCCTCTCTGCCAGGGCGCCTCTGCCAACGGCTGGTTATTTATATCCCTGAG

[0415] CAGCTGCTGACAGACAGCTGGGAGCTGGGAGGCGGCTGACAGGGACGAGCG

[0416] AGGCAGAGATGACAGGGTTGTGGGCGGCGCTGTCACTCTTAGCAGGACTGCT

[0417] GGGCAGAGCCCCTTCCCCTTCACCCAGGGAAGTCAGACTCAGACAGGCAGAC

[0418] GGGCCATCTGGGAAGGGGCACCTCAAAGAGCAGGAAGCCAGGGCTGTGAAC

[0419] CCCGGGGACGGGGAGGCAGCTGGGGTTGGAGGCA (SEQ ID NO: 50; GABA5)

[0420] GCATCTGCCCTGATGGATCCTAAGGTGGGAGGGACAGAGGTGAACCC

[0421] CGGGAACTGGGCCTCCCCCAACCTCTGGCCCTTCTCAGGTGCAAGTCTTTGCA

[0422] GTCTTGCTAGGGAACAAGCTTTCTTCCCCTCCATCCCCCCGCAAAGCCCCAGG

[0423] AGGATTCCTGCACCTCCCTGGACCACCTCATTCTCTGTGCTAATGGAGGGAGA CTGCACAGATGATTAGATGAGCGGGAACTGCCCGCTCTCCACCACTTGCCTC

[0424] CTCCTTCCTTTGGTATTTAATTATTAAAGTGGACTTGGGAGGAGCCGGAGGCT

[0425] GCGGGGGAGAGTTCCATCCCACAGCTCTGGGAAGAGGATCCACAGCAAGTC

[0426] ACAGAATGGAGAGTGGGGCGTGGGACCAGAGTGTCCGGGCTGCCCTGGGGTT

[0427] GCAAAGCCGACTTCTAACAGCCATGTGGTATGTGGGAAGGGTGGGGTGAAGG

[0428] GGCAGGGATGGGGGCACCAGGCAGGGCAGAACTC (SEQ ID N0:51; GLUT1)

[0429] ATGTCAGGGACCACCACTGTATTTCCCTGTCATCCTTCAGGATAGGCCT

[0430] CAGTAATAGGAATGTGGGAGTAGTGGCTAGGGGTCTAAGAAGCCCCCTTACT

[0431] CAGCATTGATCCTGGCCCCATCTCTTTTGCTGCCTCTACAAGATACAGTGATG

[0432] AGAACGGCCATAGCCTTCCCTTCCCTCTTCCCACGCCTTTCTGGGAAAGATAT

[0433] TTTCTTTCTCAAGATTGAAACTAGGTAGGTAGAGAACATTTTGCAGTTTTAAT

[0434] TAATACAGATTTATCTACCTCCCAGGGCTGTTTGCAAGATTAATCCCTACCCA

[0435] ATGCCTTAAGCTCCTTAGCAGACAGACACTGTGTTCTCACCAAATAAGCTGG

[0436] ACTTGTGATCTTAAGCCTGATTCTCTCAGCCACTGCCCCCTCTTCTCATATAAC

[0437] GGCCTCCTTGCTTGCCTTAAATTGGTTGGTTGGTCCTTGTGAATTTGAGTTTAT

[0438] CCTCTGCCTGGTGCAATTAAGTGTGAA (SEQ ID NO:52; GLUT1)

[0439] TTATTACTGAAGCTGTAGTGTGACTTCTGCAGCCCTAAAGATTATAGCT

[0440] GAGCAGCAGAAGGGGAAAAAAGTAGATCATCCATTACATTGAGCAGTTAAT

[0441] GATATTTTATTGCATTATTTATCAAATACTGTGGATGTTATTTGTAGCTGCAA

[0442] GCTTGACCTTAGCTAATCATAATTTGATGAATGTATATATCTGAAAGGCACTA

[0443] TGACAAAATACTCTATTGTTTTATGAAGCAGGAACATGAGCTATTATTATGAA

[0444] ATTCAATAAGTGTGTAGTTTCAAGATATTAATGCTCAGACACTTATGCACAAG

[0445] ACAGAATAGATGCTTGTAATTCAGACATAGCTGCTGAAATTTATTTCTCTTTT

[0446] AATAATATAAATATTCAAGCTATCTATCTTAAGTGATTCCTGCAAAGTCATTT

[0447] TATAAATGCAGTAAGAAGACTATAATGATAATATATTATTAATAGAATAGAG

[0448] TTGTCATTCTATCAGGAAGATGTGAAGAAA (SEQ ID NO:53; GLUT1)

[0449] ACATGCACTTGCACACACTGCACACACGTGTCCCCATGTGCACAGTGT

[0450] CTGAGACTGGAGGGGCTTCCACAGGATGCTGTTAGGGGGTCTGCCCTGAGAG

[0451] GCCAGCTTTCCATCTCCTGAAGGCCCCACCAAGTCTTGGCTGGGAGAACTGC

[0452] CCTGACGATTCCTTCTCTCCAGCCTAGGGCCCTTCCAGAGCGTGAAGTTGCTC TGACCCGTTTCTCACTTCCTGGGTGCCTCAGAGCCAGGAGGCTGGAAGAAGC

[0453] TTCAGTTAATGCTTCACCATTTTTAATTAAACATGAGAACACAGAGGGATGGT

[0454] GAGCCTGGCGTTTTAAAATATGAATTATGAAAAGCCTCCCCTCCAGATGCTCC

[0455] CCTCTCAGCTGCTTTACCCCCTGGGCTTTAGTTTGTTTGCTTAAAATTTGTTCC

[0456] TTTTTTTTC AACC ATGAAC AATATTTTCTGCC TTC GCTTAAGAAGCTGGCC AGT

[0457] GAAATTTGAATTAAATTCAGAATCTAAAC (SEQ ID NO:54; GLUT1)

[0458] TAAATATTGAGGATTGTTAATTCTCAGATTTAGTAAGCTACTTGCCCTT

[0459] ACATTTTCTCCCCAATCTAGTGAGAATATTTTGGAATTCATGCTAAGCAGCCT

[0460] GCTGACAGCAAAGAGTGTTATATTTGTTTTGATCTTCGTATTATGACGGAGAG

[0461] CTAATATGGAATGAGCAACAATTTATTGCTAAAAACCTCCCTCAGAGACAGC

[0462] CAATAAAAGAGGTATGATATCGCTCTAATGAAATAAGCCTCAGAAATGATTA

[0463] TTTCTGGTCACTGGGTGAGGTTCAGATGAGACAGATGGAAATAATAAATGAT

[0464] GTCAGACAAGATGCTCCCTACCAGATGTTGAACAATCATAAATATTTATCTCA

[0465] GAAGATAAAAGTTTTACTTTATTTTCATATTAGATATATAACCTGCTTGTTTTC

[0466] TTGCTTGAAACTGACAGATAAAATGCTGGTTGCTTTCCGGAAAGAACATGTCT

[0467] TTGAGACAAGCTGAAGGGCTACGAATAGT (SEQ ID NO:55; GLUT1)

[0468] ATATCGGGCACATAACAAGTGTTCAGTAATTGTGAGCTGCCATTATTA

[0469] TTATTACTGTTATTTTTATGATTATGATCACACTCATATCAACAAAGAAAGGC

[0470] CTGGGGATTCTCTAGTTGGAGAATGGAGGGTGATGAGTCACATTCATCAATT

[0471] CTCTGCAGAGTTGTAATATTTAGAGAGGGTGATGACATGCTGTTCTCTAGCCC

[0472] CACTGAGAGCAAGGCCACAAGAAAAATAAATCAACTGCACAGCATGACAGG

[0473] CTGGGCCTGGATCTAAGGAAGAGGAAGAGACTGATGGGGGTTTGGGGAGAA

[0474] GAGTTTGTCTTGAGTAGGGAGGGGTGACAGGCAGAGGCCACCACTGTCCATG

[0475] GCCTCATTCTCCCCTGAGGAGAATGGTTGTGCAGCACACACCTCCTGGGGGG

[0476] CGGCCAGGGACTGGCCAAGGGGCCTGTAGGATGTTTTGGTGCTGAGCCACCA

[0477] CTGGCCATGGGCAGAGGCAGGCCAGAGGCCAGGGGA (SEQ ID NO: 56;

[0478] GLUT2)

[0479] TAAATCTGAGCCTTCATTCTTGGAAAGGCATGGTAAAGAGAAGATACC

[0480] AGAATGGGGAAAGAGCTGAACTTTCAAACTAGAGGAAACATCCTTTCAAATT

[0481] TAATTCACAAACTGGGAATGGAAATGTCATTGCCTCACTCAGAGGGTTAATC ACTTTCTAATTTGAACTGGCAAATTTTATAATGACACTGTGAAATTAGCAGCA

[0482] TTTTATTGTGTTAATTATCTGCAGGACACTGTATCATTACTTCCAATAAATGTA

[0483] AATGAGATTGTGTCTCCTAGGACAATACACAGACAAGGTGAATTTTTATTGG

[0484] CTATTTACAGGATTCCAGGACAGTCTTTCTTCTTTTTTCTCTCCTTATAAAATA

[0485] CTTTCAAACACTGTCAGCTTGCCATGCGAATTCCCCTGTATTTTATTTAATACA

[0486] ATATTTTAAAGGTCATTAAAGTTCTCCTTAGCTCACTGGAACTCATTTTCACTC

[0487] AAAGCAATGTTGATTATTAAAAACCAT (SEQ ID NO: 57; GLUT2)

[0488] TGCCTGTTTCACTAGTAACTGATTTTGAATTCCAGATAATACTATTATG

[0489] TCCATTCCTGGATATTAGGTCAAACAGCTAAACTATTGCTTATTCACTTATGT

[0490] CCCAGACATGAGGTATCACTAGATCCAGAAATGAATTATCCCTGGTGGCCTC

[0491] TCTTTGACACATGGCTCCAGGAACTCATCATTTAGACGTCAATCATTCCCTCC

[0492] TCCTGGCTCTGATGTCTCACAGGTTTCCTGACAACAATTAGGGGGACTAAAAT

[0493] CACTCATAATCACAATCACACCTGCCTTCCTTATTGGCTCACAGGCAATTTAC

[0494] TTCTCTCAGCTCTTCTATCACATAGCAGATCCGGGTGTTACTTTTCCAAGCTTC

[0495] AGTTATAGACATTCAATCTGACAGCTTCCCTGAAGTTATTATTCTGTCCTGTA

[0496] TTTTCTGATATTGATCCCAAATGTTTGGGTAAAAGTATAAAAACTGCAGTCAG

[0497] CCATTCAGATGTTTAGGAATATCCAGA (SEQ ID NO:58; GLUT2)

[0498] TTATGTGATCATTTACAAATGACTGTTCCAAAATTCTTACGTTTATATA

[0499] TTTATTGCAATTTTAGGCTGTGTTTACGTTATGAAGACATAACTCCTGTGCAT

[0500] GCTGTTGGGTTTTATAAAGGCTTCTTTCAGACATTTGCATTAGGAATTCAGTT

[0501] TTGAAATAAAAGTCATTCCCAAATAAGGCTGATGTTTTTTATTAAAATAAAAT

[0502] TTATGCCCAGTCCCCCAAATGGCTCTGGTGGTAATTAAGCAGAATCAAGTATT

[0503] GGAATGTAGCAATTTTTAAAAACCAAGCTGACAATTAGTTTTTAAAGAGAGA

[0504] CAGTCACCTAATAGAGATCAAATTTTAATGCATTTTCTTAGCCTGCTGCCATT

[0505] TCCAGAACTTTGTCAGACATTTTCTGTTTTCCAAGTAGAAGAAAAAGTGGCAC

[0506] ATTACAATAAATTTTTTTCGTTTTTACAGTAGTATTTCTTTGCTGAAACATGTA

[0507] CTCTGTTGAGCTTATTTACATGACCTC (SEQ ID NO: 59; GLUT2)

[0508] GCAAACATAACCAGAAAAAAAATTCTAAGGTATGATAATCATTACTAT

[0509] GATGATGATGATGAATTGACTTATTGATGGACACATTAATACCAGTTGATTTA

[0510] ATCTCCAACAGAAAGGAAAAGTGAGTGAGTTGGAAGCTACAGTTAGGAATA AAAGGAAATTTGGTGTCATCATTGCTTTTCCAGAGGACTGAGAACGTCTGGG

[0511] GACACTTAGTGTATAACAAATAAAAAACCCTTTTCCCTAGCATCAACTGGGA

[0512] TTATAAAACTCACCTAGATGCTCGGGCCTCGTTCTTTAATAATGAACCTTGGA

[0513] GAGGGGTTGGCTGCCCCAATCCTCAAGTGCTCTTTTTGTGCCTCTGCCAGACC

[0514] CATCACCCTGAGCCAGCACATACAAACTGTCGTCCTGATCCAGAGCAGCTGT

[0515] CTAATTACAATTTTGTTATTATTAATTACAGAGTAGAAAATCATTTGTAATTA

[0516] GCTCAGCCTAAATCATAGTTTAAACAGCCAAAA (SEQ ID NO: 60; GLUT2)

[0517] ATACAACACTTCATCATTTCAGTTACCTCTTTGTACAGCATCCAAGATG

[0518] CCTACAGGCCTAGATGGAGCAGCCTTATTAGAACACCAAGGGGAAGGGGTAT

[0519] CCCTAGCTGGCTAATCACAATACTGTCAAATTGCTACATCAAACGACTATAAT

[0520] TCACTGAATAGGGAAGTTATTAAAATTCCAAGTTCTCTCTATGCTGTGACCAG

[0521] AATGACAATGGCAATACCGTCAGCCTTCCTTGCAAACAAGGAAACACATTAG

[0522] GAAAGGTGTAGGTCCCACAATCTCCTGTATCTATTCCATGAGAGCTACCCTCA

[0523] ACTAAACAAATAACGTGATCAGTTTTGTACCTTCTGTTATGTGACATCATTTG

[0524] TGCAGAAGAAAATATCCCAATTCTACTACATGTCCCATGAAAGTTAAGGTCTT

[0525] AATATTCTACAACTCATAGCACAGAACTCTGCACTACAACCACCCTTCACAAT

[0526] CTAGAAGAGCAACAGCAAATTGGCTAATT (SEQ ID NO: 61; GLUT2)

[0527] ATGCATAGAAAATCCAAGGAAGAAATAAACACCAAATTATAAAAAAA

[0528] AAGTAATTAGATTACAGATGGCAGTCCAGAAAGAGCAGGTAATTTTACCAAT

[0529] ATTAAAAGCAATTTTTGAAAAAGAAATAAAATCCAAGTCTTCTGGTACATAT

[0530] TGTAGAATTTTTAAAAATAAAACTGTGCCACAATAATATTAAGGAAAATAAT

[0531] CTTAACAACAGCAGTAGTTGGCTGAGTGCAGTGGCTCATACCTATAATCCCA

[0532] GCACTTTGGGAGGCAAAGGCAGGAAGATTACTTGAGGCTAGGAGTTGGAAA

[0533] CAAGCCAGCCTGGGGGACATAGTGAGGCCATATCTCTACAGAAAAAAAAAA

[0534] TTAGCTAAGCATGGTGGTGTGGCACTGTAGTCCTAGCTGCTCAGGACGCTTAG

[0535] TCAGGGGGATTGCTTAAGCCCAGGAGTGTGAGGCTTCAGTAAGCTATGATTG

[0536] CACCAGCCTGGGTGAAAGAGCAAGACCTTGTTTCTCAA (SEQ ID NO: 62;

[0537] GLUT3)

[0538] CACATAAAAGAGAATTTTTTGCCCCACAGCACTTCTTGTCCATTAGAG

[0539] ATATTTGCCAGATGACATAAGATCTTCTACACTGGCAGCTTCAGTGGCTATTT GGAAAGCTTCGCTGCCACCTGCAGCTGCACCTGCTGTGAGATTTTACTTGTAG

[0540] ATTCCTTGGTGAGTACAAAATCTATTGCCTAGATTAGTAAATGAACCTAATCC

[0541] TAGCAGCAATTTCCTAACATAGTTACAGAGTTTCCCTGGGTAGGATTGGACTA

[0542] TGCGGATATGTTTTCCCTTGTAAATTTATATAAACTAATGGATGCATAAACAA

[0543] TAAATGAATTCTGTATAGGCATGTAATTTTCTTCATTAACTAACATTATTATA

[0544] CTAAAATTGCTTAGGTCAGAATTCACCATGGAGCTGGTCTTCAAGCTGTGAGT

[0545] TTAAAAAAAGAAAAAGAAAATAATTATCTCTGAAATTGTCTGCTTTTTTAATT

[0546] TAGTAATTTTATCTGCTACTTGCAGAAA (SEQ ID NO:63; GLUT3)

[0547] AGTGGACTCTGAGCTAGAGCTGGCAGAGCAGGAGTTGTTCAGAGGTC

[0548] CCACTGGTCCAGGGAGGTACTGTCCACAAAGCATCTCAGGTCCGTGTTTGCAT

[0549] CATGACCATCTCTTGGGAAGAACTTAGCTTTAAAAGTATTTTTCTAAAATACA

[0550] TAGGCTGATGACCATAGTCACTTAAGTTTCTAAGGAAAATATGAAACTTACT

[0551] ACCTAAACCTGTGAGTCAATTGAGGTCTTTTACTTCCCAGGACTACAATTTTA

[0552] ATTTCTCTGCAAAAGGATCTGTTCATTTGCATGCGCTCATGTGATGATTTTGCT

[0553] TTGCTTTAGAGCTGTTACTTCTCTGAAAATAAAAAGATCTCTAAAGCCTCTGA

[0554] TAAAAAGAAAAGAGGATAATCACTGAAAACTTTGTCTTAACATATCTATCAA

[0555] CTTTATGACACAAATTTTAAAAGAGGAAAGAATTTTGGGAGGCCGGTGGTTT

[0556] GTGCTGATAATACCTGTAATTTTGTAATGTG (SEQ ID NO: 64; GLUT3)

[0557] AATTCACAGGCTTTTTACCATGTATAGAAGGTGAGGTTTTAAAAAAAA

[0558] AAATTCTCATTTTCTGCAAAGAAAGGTATTTGAGGGGAGGGGAGATGGTTTC

[0559] CTCCTGCCGTTGTGGGCGGCCCCGCACTTCCTGTTCTTCTTCTCATCTCTCCAT

[0560] CTGTTCGGTCCCCCTATCTCGTGTACCCATCGTGTAAACCTGAGCGAGGCGTT

[0561] CAGCCTGAACACTTGAAGTTTTACTCTTTACTGTTTAATTCTTATAATGTTCCC

[0562] TAAGAGATGATTTTCTGGGTTACATGGCCTGAGCATAAGTTAGACGTGGGAA

[0563] GAGAAGGGCCAAATTACAGCAGGAAGAATTCTTAAATTGGATGAAATTGTAG

[0564] CTTTTAGAAGCATAGCAGAAATCTGAAGAATTGAAGAAGCAAATTACGTAGA

[0565] ATTAACTCTTAGTGCTTTTAGGTTGCTATTTTAGCAGGAAGAAATGAGGTCGC

[0566] ATTAGATGCATTACAGAAGGAGACAGAAAT (SEQ ID NO:65; GLUT3)

[0567] GACTTGCTTATATACTGCCTGGTCTGTTTGTTTGTTCTTGTGTGTCTTCT

[0568] TCATCTTCCAAATAAAATGTAAGTTCTTGGGAAGAAACCTCAAACTTTTATTT TCCCTACAGCATCTACTGTGAAATTCGTATTCAAGATAGGATGAGAGTTTCTG

[0569] ACTTATAATCAGGTTACTTGCAAAGGTTGATTTTTCAGCCAGTTATTTGGATT

[0570] TTGTTTCTCATTAGGATAAGGTTGTAAAGGGTAGTTATGTTCCCATACCAGCC

[0571] CCCAAATGTCTATTTAATTGGCAATATAGCTGAGTAGTACTACAAGACCTAA

[0572] AATCATCTACAACAGTGCTTTTATGGGGAAATGCATTGCAGATTCCAACTGAT

[0573] GATGCCAGGAGCTCATTTTTCTGGCGTTGGTTCCAGGACAGGCCTTGACCCAT

[0574] GTGCAGTTAGAACTATCCTCCTGCCTCTACACACACACATACATATATTAGAC

[0575] TGGCTGTCTTGAGGTGCATGTTCCAAA (SEQ ID NO: 66; GLUT3)

[0576] TCGAGAATTTACTACAAAATATACTAGTGTGAGGTGTTTTACAGAATA

[0577] CTCTTTTCCTAGACTCTTAAAAACAAATCCTTCTGTGTTTTTCCATTTCCCATA

[0578] CAAAGTATAAAAACTTAGAATGGGTGTTCTGAAGTTATTCTGTTCCATGTTCC

[0579] TCTTCTTAGCTCAAAGTGATTAACAGATATTACTGTAACTAATGAGAACACAT

[0580] TGTGCAGGTGGGAGAATATTAGATAGAAATAGAACATGAATTAATGACATCT

[0581] GAAAACACTGATGGATGTTCATCAGTTGCTAGGGAAACTTTAGTTTCAGTACT

[0582] CATGCCAATGAAACAGGAATGCTGTCTATGCACAAGGCAGCTAGGATCTATC

[0583] TCAGAAATGGAAATGTTGTGCCTGGGAAATGATGATCAACAGGATAAGGCTG

[0584] TGAATATCAGGATCTTCCAAACAACATAACCCAATAGCTACCCAATATGATG

[0585] CTGATTGTGTTACCAGAACAGGGTCCTGATC (SEQ ID NO: 67; GLUT5)

[0586] GATAGTAATTTCTTCTCCCTGTCCGATCTGATAGCCATCTTGGCGTTCT

[0587] CCAACACGGCTATCAGACCATTCCCATGCCAAGCCTGGGAAGGACTTGGTCA

[0588] ATACATCCGCTGTGTCAAGCTTTGAGTCATTTTTATGTTTGTTTGTTCCCCTTA

[0589] TTACTTTCTTTGGCTGAGTAAAATCTCTATTACTTTGGGCTCAGGAACGAAGA

[0590] ATGTGCTATGAACACCCGAGGCGTACATTTTATTTTATTTTTATTATTTTTTGT

[0591] GATGACTTCATTATTCCCTGTTGCCAAAGTTTTTGCCAAAATTGTTTTTCTCCC

[0592] TTCCTCTCTCTTCGGCATCCTCTTTTCTGGCATCTCTGCTTATTTATTGCAACC

[0593] GCACAGAAGTCGGCCTTGCCGTCGCATGGCCTGTCTTCCTCGGAGCCCCAAG

[0594] TGGCTCCCAGATTTCTGGGTTTCACGAACCAGTAAAAAGTCCCCTCTTCCCCT

[0595] TCCCTAAGAGAAGGGTTTAGACAAG (SEQ ID NO:68; GLUT5)

[0596] ATGAAGCCTAAAATAAACAGAAAGAGAAATCTGTTATGAAAGGGCAA

[0597] TAGCTCGCTAAGCTACCTATTATAAAATAGTAAAGGAGTCGGTGATTCAGGG AGTTAGTGGAGACCACTCCAGAGACATCATCCCACTGCACTTACACATCTAA

[0598] GGAAAGAGATATTTTAAAGAAAATAAAAAATAATGTAATCTTACAAACAAAC

[0599] CACACCAGTTGGCACATATCTTCCAAATGTACCTTCGAATTTTCAAACTCTAC

[0600] ATATTTGCTCCTGTATTTTCCTATACTTGGAATTTCTTCGATTTTTTAAATGAG

[0601] TCTCAGATGATATTTAAACTCCTCCATGAATATTTTCTGTTTTTAGCAGTCTAA

[0602] TATTATACTCTAATCTTTCAAATTTCTAGAATTTACTTGGCTCTCCTCACACAA

[0603] ATTGCAAAAAGCCTCACTAGCCCTTAAGAAATTATTTTCATTTCTCACTTGCT

[0604] TTTGTATTTGTTTGAATTAACATTTCCTC (SEQ ID NO: 69; GLUT5)

[0605] TTTTTTCTTCCTTTTTTCCTTTTCCTCTCTTCTCTCTGTCCCTCCTTTCCCT

[0606] TCCCTTCCTTCAACTAGGAAATATTAAGGGTTTTCTGTGTATAAGGCCTGATG

[0607] TACTCTAGATAGTCACAAATAAAACATACTTTCCCAACTCATGAATCGTGTAG

[0608] GCTAGTAAGGAAGATTCAGTTAAACAAGAATGTGATGAGCTTTAAGACATAA

[0609] GTTTTAAATATAAGGAGTTAGGGAAAAACTTGGCAGTTGGACTTAATATTAT

[0610] CTGTGGGACAGGTAATGCTTCCCTGCATAGAAATTAACTAGAAGGATTTTAG

[0611] GGAATTGTGTAGATTGGAGGAACAGTATGTGAGAAGGCCCAGGAGCTAGAA

[0612] GGAACATGGTGTTATTAAGGAGATAAGATTCTATGACGGGAACAGAATATGA

[0613] GGGAGTATGTGTAAAGAAAGTTGGGGGAAGATGGGGCTGGATAATGGAAGA

[0614] TTCTCAAAGCTATAACAAAGAGTTTGAATAAG (SEQ ID NO:70; GLUT5)

[0615] AGAACACGTGGATCCGTTAGAGGATGCCTCTTGGTTGCTTAAAAAAAG

[0616] AGTCAGTCCATTTAAAATTGTTAGTTTTCTATAGTCTTTCAGAACATAGAATT

[0617] CATTTAGAATTTCCAGCAAAAGTACAGAGAGTCTTCAGGGAAGTTGAAGAAG

[0618] ATAAAATAATTCTGAACACACTGTACACTTTCAGAAGAAAAATATTAAGAGT

[0619] AAAAATAGTTTCCCAACTCAGTCATTCTTTGGAACTGATAATTTATTCCTGAT

[0620] GGTTCCCCAAAAGAGGGGAAGAGAAGCAAAATAATATATTTGGCTCCTCTAA

[0621] TAGATATTATTCATAGCTCTTTTCCCTCATAGGATAATAGACTATTAGAGCTT

[0622] CTATAAAATCCCTTTCAGTAATTTTATGGTGGGTCTGCATTTTGTAAGCCATG

[0623] TTTTATAACTGAGAAAAGAAAAAAAAAAAACACAACCACCAAGAGAATAAT

[0624] TTTTACAATAATTCATTCCACTGATTTTATTAT (SEQ ID NO: 71; GLUT5)

[0625] AATTGGAATCTCTCCCAGAAGCGTTTCCTGTGATGGTCTGAGCCATAC

[0626] CTGTGCCCGTGCTATGTGCAGTTGGAGAAGTCAACAGCCCCTCCCACCCCAC CACCAATTCCCTGAATATTTGCAAACTGTGTTTAGAGAGAAAGAGGAAGCTT

[0627] TCTTGATAACTTCCTTGTTTATACTAATAGCAAAACTCCAACAACTTTGATGG

[0628] GATCTGCACCAGCAATCTTTCCCTAGGGCTGATGCAGGATATTTAGGGAAGC

[0629] TCTGTGTTCTAGGCAGGTAAGATAGCCAGGTTCCTAACGGGAATCAACATAT

[0630] CTCTTTAATTCATATTTGAAGTAAAAGAAACATCTCTCTTCCTCCCATTTGGCT

[0631] CAGGAATTCATCTCTAAAAATCTATCCTGAGACAAAATTTTATGCCCAAAGAT

[0632] AACTCATGCATTATTTTTAATAGCAAAAAAAATGGAAACAATCTTTGCCCAA

[0633] CATTAGCAAGATGATCGAATGAATTATGGCAT (SEQ ID NO: 72; GLUT5)

[0634] CACATATTGAACAGGGAACATGCTGGAGCCACCTCTGCAATCCTCCTT

[0635] CCCCACCTCTAATTCCTCCCAGCCCCAACCCACTGCCTGCTGTCAGCCTTCAC

[0636] GTATGGAAGTAAGTGGAGTGAAACCTGATTAAAAGCACTTATGAAAACTCAG

[0637] AAGTGAGAATGGAGCCAGGATACTGCAATAAATAAAGAATATTACCTTAGTC

[0638] GTGTTCTTCTCCTTCCTACCTACTATTTAATCGCTGCCATTCCTTCAAATTACA

[0639] GACAAGTGGATAGACTTAAAATACTCAGCTCTTAGACTGTATCCCTAACAGT

[0640] AAAGAAAAACTGGTCTCATATTTGAGAAAAAATCATAGCAGACAATCATAGC

[0641] TTTTCTAGCTATGGTACTTTGCCCACATCCTTCAGAGCACTACCAGTCTACCA

[0642] TTTGCCAGTATCTTTAACTGTTTTGCTTAAGGCCTCCTCTGAATTTACAGGAG

[0643] GCTTCCTGGCTATGGGGTTGGGGCGGGGCAG (SEQ ID NO:73; GLUT5)

[0644] GCACAGAGATTATAAAAGAAAATATTCAATAAGAAGAGATAACTTTA

[0645] ATAAAATATACTTAGATGCCTACATCAACTTCTAAATTAGGCTTTCTTGTTTC

[0646] ATTTTGCTAGGTTTTCTTTCCGTTCTTTATGTAGTTATCCAGCTTTCAATTTCCC

[0647] TCACCCCCTCCCAAATACGAAATGCTTTAAAATTAAGCACAAGTAGAAACTA

[0648] CTTTAGTATATTGCATTTATTACACATCGTCTTTTCTGAGCAGAAGTTTGTAGA

[0649] ACCAGCTATTATACTCCATCAAAAAATATTTGTTTATCTTGGAGCATACCCGT

[0650] GTTCTTTGCAAGGCAGAGAAAATGAAAAGAAAAGAGATTACATTTAAAATAT

[0651] GCAGAATTTGATTGCTGGAAAAGTGCTTTACTAATGCAGTTTCAATGCATATA

[0652] AAGAAATGCATTTGCTATTCATTTCACTTTCTAAAAGGGTGTAGACAGTTTAT

[0653] ATACCCCTGGAAAATATTTTCAACTTAA (SEQ ID NO: 74; GLUT6)

[0654] AGAGCCTTCAGGTGTCTCCTTCTTACTCAGAGTTGCTGAAAGAAGTAG

[0655] GAAAACACATGGGGTTCTATATACGTTTTGGGGATTGCTGATGTGATTTATTG AAGATTTTCGTAGCCTTGAAAAATTCAATTTTATACAAAATCAAGTATTTTGC

[0656] ATAATAAAACTGGTAGAAAATGTGCATATGTTTTACTTTTATGTTCCGCTTTT

[0657] ATGAACACTGTCAGACTGAAAACTGAATTTATGAAGAATTTACCCAACACAC

[0658] CAAGAAAATTGCTGCCAAGAAATCTTGTATTTACACAGTGTGATAGAAAGAG

[0659] GCTGAGAGGCATTTAACACTGCAGTCCAGTTTATATTCAGCCACACTCTATAT

[0660] TTAACTTTTTCTTTTCCAAAACACGAGAAGCTAAAGGAAAAAATCTTTATAGA

[0661] AGAAAAAGGCCTTCTTTGAGATAAAAATTGTATAATTCAGAATAAAACTGCA

[0662] AAGGAGAGAGAAAGAATAATCTCAAGACGAT (SEQ ID NO:75; GLUT6)

[0663] CCGAATACAACTAAACATTCCTTTCAAATAATGTCACATTAAAACTTG

[0664] AACAAGACTCAAAGAATTTTTCCATATTCCAAAGAAGAAAGCAATATATAGC

[0665] AGCCTCAGAAATACTGAAATTGCTAGTAAAACAAATCTACAAAATTGACTTT

[0666] GGAAAAAACAAACACCAAAACAGCAATAAAAGCAACAAAAACAGCCAAGTT

[0667] TATGTACAAAGAGAAGCAACCAATATTAAAGTAAATAAAATATTTTCCTATA

[0668] ACCAGACTTACTGCTATGTTCCACGGGTTCAAATGAAAGTGCTTTTCTGCAAA

[0669] ATTATAGTTTACAGCATCGTATGTGATTAAAGAAGTAGCAGTGAAAATGTCT

[0670] ACCTGTCTACCGGAAGAAACGAAAGCTAGGTATGCTATAGAAAGGATAATCA

[0671] GCTATTTCCTGTTATTTTCCTCAGAACCTTAAGTCAACTCAATCCGACAAGCA

[0672] TTAGCTTCATTTTCCTTTTCTCTGATTCAATTCAC (SEQ ID NO: 76; GLUT6)

[0673] GTGAGTTATTTTCCTCATTTATCTTCTCTTGAACAAGAGACATTTATTT

[0674] ATCTCCCCCTTCCACTCCCATGCATACACATTCTCTGCACATTCATAGTGCTTA

[0675] TTTCACTTCATCATAATCACTTGACCTATTTTGAGAGAGGAACATGTTTTGTT

[0676] GGGAATGCAAACGATTCAGTGCAACATCTATCATATATACACTCAATGAAAA

[0677] TTATGCTTATTTTTTTCCTTACCAATACAACCCCCTTGGAACTCTCAGTTTTGC

[0678] CTTATGATATACCTGCATTTTAGAGTGTGATAAAATTACCCTAAGAAAACATC

[0679] AGTCTCCTCATTTGCAAACTTATGAGTCTGTATCATAAGACTGGGAAGAAACC

[0680] GTTTTGTTATAAAATTCTATGATTATATAAAATACCATAGTGTTATTACCCTTG

[0681] AAATGATTTTGTTTGCAGTGTTTGCTTCACGTAATTTCCCTAATAATTAGCCG

[0682] AGTCTTATGTTTATCGTACTTTTTT (SEQ ID NO: 77; GLUT6)

[0683] GAAGATAGTGTTTGTCAAGGATCAGCAACAAAGCCTGGTACATAGTA

[0684] GGCAGTCAATAATTACTATATCATATTGATTCTAAAATGCACACTTATTTTCA CATTTTAACATCTCTGAAATTGAAATGGATCTATATTGTGTTATGGTTTAATG

[0685] ACAGTTTTTTCTCTTTCTTAATGTGCATAAAATAACAGTACATCTTATAGTCA

[0686] GTGGCATCTTAAGTTGATGAAATGCAGTAATTTCTTCTCTCTCTTACTACTGTA

[0687] TCCAAGAACATATTTTACTCCCTCACAAACCTGACAAGTAACACAGAAAAAA

[0688] TAAAAGCAGCACTATAACAAGTGGTGCCTTACAACCATTTAGAATAATCAAG

[0689] AACAATTTTTTCCCATTTCACCTGACTATTTCACTTTTAGAGCTGTGATAAGA

[0690] GCAACTGAGATTTTTGTGACATAACAGGATTGTTTAAAAGATCTCTTTTTGCC

[0691] TTTTTCACAGACAGGCCTTGTTTAAATTAA (SEQ ID NO: 78; GLUT6)

[0692] ATGTAACTCAGCTGAAGAATCATTATAGGATTTTTTTAAAAACTGGAA

[0693] ACTTCCCCCCTGCCCCCAGCTGTCTACAGAAATAGATTATTTCTTCTAATTTCC

[0694] TGATTTAGTTATAGTCATAGTGTACATAAAAACAGATTTGATTTTCTTGGAAT

[0695] CAAACATAAAATGAAAGCCAAGGCCTTATCATTGAAACAAGACAGCAGTTTT

[0696] GCAAAGCTGTGTACATCCTTTGAACTCAGATGACATTGTCTCAGAGGAAAAG

[0697] ATGTTTGATGAGTTTTTTACACTTTAAAGCTCTTCCTAAGGAAAATGAGCTGG

[0698] GAATCTCCTTTGGTAGAATTGAAGGGGGGTGGATCATTTCGTCTTCAAAAGAT

[0699] GAGGGGAAAAGTGTCATTTTCTATTTCCCTTAAGTATCATTCTTTCAAATACT

[0700] TGTGTGACAAGAGGTTCAACAGGCTTAGATTTGGGTTTTTAGTTTTCCAGAAA

[0701] TGCATGGAGGACTTAAATGGCCCTGTACA (SEQ ID NO: 79; GLUT6)

[0702] AAGATTATTGGCTTTCTGACTAGTGCCTTAAAGCTGGAGTTGGATATA

[0703] TCAAAGCGCTGTTGCATGATTATTTATGATTTCATTTTACTGCAGTGTAAATTT

[0704] TTGTAATGTCAGCATTTAAGAAATATTATTCCCGTCAACTTTACCAAGCCCTG

[0705] AGTGGCTTCTTATATATTACTTTTCAGCATATTTGCTAATTTCCTCAAGCAGTG

[0706] CTAGATATTCACTTTGGAATTTGCCTTTTGATGTCTCTTCTGTGATGAGCAGCC

[0707] ACCAAACAGCCCCCACCACATTCTAATTCTTTATTGCAAACAGTAAAATATGG

[0708] AGCACTGCATCTGAAAACCACTGTGCGAGACAATCTCATTTTTACTTTCTTTC

[0709] TGCTTAAATATGCAATAAATATGCATTCTGATAAGTACAGTTCTAGAATAAG

[0710] ATAGTTTCGTGTAACATCTTGGAGTAAAAAAGAGAAAGAAAAATTTTTTTTTT

[0711] TTGAGAAGGAGTCTTGCTTTGTTCCC (SEQ ID NO: 80; GLUT6)

[0712] CTGACTGCAAACATGTCTATGCAAGTTAATTTGCTTTGTGTGTCTTCCT

[0713] AGATACTATTATCTAAGATCGAATTCAGCTCTGTGAATAGTTTTCAGGCCATA TAATTATTTTGAAGGTGTTAGAGGCATTTCAAATGAAAATTTTGCAAATGAAG

[0714] TGACACTGGATTGTGTAAAATCCTATCTTTCTAGGCTTGTTGTTTTAATCTATA

[0715] TTTTAGTTGGGTGATAAGTAGGAAGATTTTGTGCTTTCAATAAACACTGCCTT

[0716] GTGGTTTTATTTTTAAAACTGCATCTTTAAATCAAGATGAAGCATATGGAAGC

[0717] TTCATTGCTTAAAAGCTGCACTTTTCTTCAGGGCAAATCTCAGAATCATAGTC

[0718] TTGACAAAGAGCTTAAAAACCTTTCAGAAAAAAAATTTAATTTTCTTCTTTCT

[0719] AGCAATTCAGTTACATTCACAGATTATTTTATACAAGAAAACAGTAGGCATTT

[0720] TTTCTTCATATCTGCTCCTATTCCTC (SEQ ID N0:81; GLUT6)

[0721] TTCTCATAAGTAAAAGCTTCTCTTAAAATATTGGAGTTCCTCCATATGG

[0722] GAAGCTTGGGGAGAGAAGAGAAAAAATTAAATTAAAGCATTTGAAAGTATA

[0723] TTATAGTTTTATCTCTTTGGCTCTGCGATTTCAGAAATAATATAATCCTATGGA

[0724] AAACAGGCTGCCAGAGGATTTTGTATAGTGGCGCCCCGACAGGATGGGACTG

[0725] AATGATGTCAAAAAAGCTTATATAACAGTTTTGTTTTATTCATTTACATAAAT

[0726] CACATTTACCGTAGGTGTTTTATTCAAAATAAAGTTTGGGAAAAATAAATTGA

[0727] ACAGATGAACCTAGGCAAACCTGGTGATTTTTTCATGAGTAGTGTTTGATGAT

[0728] AAAGGCATAAAAGCATAATGTGGTATCATTTCACCAAGGCACAATATGGATC

[0729] ACCCATATGGAGCAGAAGTTGATAATACTGCAAAGAAGCAAAAATAAACAG

[0730] AACAGATGTACTTGTAAAATCAAGTCTGCACC (SEQ ID NO: 82; GLUT6)

[0731] TCCTTGGTGAACTCATCCCACGGGAGGCGGGCTTCCTCAGGGAGCTCC

[0732] TTGCTGGTCAGAGGCAGGGTCACAGCCAGGATCATGCGAATCCACCTGTGCC

[0733] CTGGGTGACTCATCCAGCTGGGGGTTTTCTGGGCAGCTCAGCCTGGAGGAAG

[0734] AGGGGCTGCTAATGGCCTCGCCAGAGGCTGACACCTGCAGTTGGGAGGGGTG

[0735] CGGACGGGAAGTTGGTGACTCAGCAGCAGGCCCTGGACAGCTGCCCCTGTTT

[0736] CCTGGCCCTGAGTCAGGCTCATCAGCAGCACCCAGGGCCATATGGGAACCCG

[0737] CCTGGCTACCAGGATGTGGGTGTGCAGGAGGACAGGTGGCTGGGGGGCCAC

[0738] AAGGAGGGCCAGTGGGAATTGAGAGAGGCGCTTCCACCTGGGGGCCTGAGC

[0739] TGTGCAGGAAATGCTCTGACCTGCTTCTCAGAGACCCAGAATACTTTTTTTTT

[0740] TTTTTGAGAAACAGTCTCACTCTATTGCCCAGATTGG (SEQ ID NO: 83; GLUT7)

[0741] TGTAGCAGTGGCTTTCCAGGACATGCCCCAGGAGCACAGATGCTGTGA

[0742] GGCCGGGTTGCCTTGCCCTCGTTGCCACTTTCTTGGTGACTTCAGTCCATGGG AGGACCCTTGGCACACAACATCCCCAACGAGGGACTTGACTCTGGCACTGGG

[0743] CTGGTGTCAGCAGAATGTGGCCCCCTCCTCCACGCTCCATGAGGAAGAGGTG

[0744] GCTGCCCTGCCCAGCACGGGCCAGGCCGTGTGGAGGAAGGAAGTGCACCCA

[0745] GGTTGGCTGTGTGTCCCAGTTTCCTGCTGACTGCGCCCCGTCAGCACTCCTGC

[0746] CCTGGAGGTGGGGCTGCTGCTGGGATAAGTGGGTCCAAGGTTCTGGTGACTT

[0747] AAGTTGTGACATCCCTGGGGATGAAGGGCTACAACAGTCTTTCAGGAGGCTC

[0748] TCTGGGGACTTGTCCCACCCAGACCCCTCTCTGAGAGTGGGGGAGGCCACTG

[0749] GATGCCTCCTTGCATTGGAGGCCCCTTCCCCACCG (SEQ ID NO: 84; GLUT7)

[0750] CTTTGGAAATATTCTAAGTCTGGTAGAATTCAGTGTGGAGCACCGTGG

[0751] TGCCCACATGATGGATGCCCTGGAGTCTGGGTTGCTAGTGTTTTTTTGCAGCT

[0752] TTGTCTGGTCTCGGCCTTTATGGCAGAATTACATTATTCAGCCGCAGCCTTTC

[0753] CAAAAGGTTGAGTCAGCCTGAGTTTGCTGCTGGCGCCGTGGAGCCCTAATAC

[0754] TTTGCTGAGCCCTCGCATAGCGCCGGGGAGGCACACCTACTGTCCACCTGCTC

[0755] CCATGAGTCATAAAACTGCCACACTGCTGCCAGGCCTGCCAGCCCTGCATTTT

[0756] CCTCCAGTTGCCACCCTGATGAGAATGAACTTAGCCATATAGGTCAGGGCAC

[0757] CAGGCAGCACTTTTCCCCCAGCCTGCCTTCCTCTTGCAATGTGCTATTGGACA

[0758] ACCAGTTCGTAAAGCAAGGGCCAGGATGTTTCCATTGCTTTATACCAGGTGG

[0759] CTAAGGAGAGAGGGCAGCAGGGCAGCCTAGA (SEQ ID NO: 85; GLUT7)

[0760] CCACCACAGAAGGAGCCCAGGGTGGAGCTGGGGGGGCATGAGGTGAT

[0761] GAGGCGCCAGAAGGAGCCCGTCAGCACTTTTCAGTGCCCAAAATAACAAAGC

[0762] AAAAGGAAGCACGCAGGGGTGCAAAGGGGCAGGCGGGGCGAGGGGCTGTGC

[0763] CCCCACACCTGGGAGGGGTGGCGGGGGGAGTAAAAAGGCAGGAAAGAGAGA

[0764] GCAGAAGAGGATGTTCAGAAACAAGCCGCGGAGCCCGGGTTGGGCTGTGGT

[0765] GAGTATCTAGGTCACCAGGGAGCCTGCAGGCCTGACCACAGGGAGACCTGTG

[0766] TTCTCAGCTCTCCTCTTCCTCCGACCCTCCCAGAAAGGTGAGGTGACCCCAAT

[0767] ACAGCCTGAGGCCCTCTGCGCGACCACCCCCAGCCCCAATATCTGGAGGCCA

[0768] GAATCCCCAGCCTAGACTCATTGCTGGAATTTTGCCATGATTCAAAGCTGATT

[0769] TAGAACTTAGAAGATTTTGCTTAGCAGATCGAAATGGC (SEQ ID NO: 86;

[0770] GLUT7) CCTGAATCTTGAGCAGTCATTATCCCAAGGCCTATGAGATGTTCCCAG

[0771] TATAATAAACAAAACCCTCTGGGATGCTGGCCCTGCCTGCACATTCCTGCCCT

[0772] CTCACTCACAGCTTTCCTCTCATCCCTCCTTCAAAGCATCTGAGCTGGACAGC

[0773] TACCCAGAAAGCCTCCCACTGCTGCAGAATTGGAAATGTGGGTCCTCCACCT

[0774] AACAGCTGGGTTGGATCCTGGGTCAGCAGCAGATTAAGGCTACACTGCAGGG

[0775] GCTTGACCCAGGAGTGTATTTAGGAGCTTCTCGAGTCAGGCAGCAGGAAGTG

[0776] AGTCATCACTCGCCATCAGAGATGCAGGACGCCCTGCAGCTGGACGCTGCCA

[0777] CAGCCTGTCAAACCTCCTAAAGGCAGCTGAGTGCACTGGAAAGCAGAATGGA

[0778] AAATTACACTCACAGTCACCCCTTTTAAAGAAAGTTTATGAAACTGACCAGT

[0779] ATATGTTGTGGAAAAGTCTGGGCTCGCAGAAGAG (SEQ ID NO: 87; GLUT7)

[0780] ACCCGTGAGAAGTGGGGCCAACTTGTTTTGGAGATTCCGGGACTTGTC

[0781] TTTGGGCAAGTCTCCTTCCTAACACCCCCTTCCCCAGTCTATGGGGACAGATT

[0782] CAGGCTTGTGTGGCATTTGAATACTCTTGATCTGCCCTTATGTAATAAGTGTG

[0783] ACTCATGTCCCAGTATGAGTCACCCAATCCCGCGCCCGGGGCATCTCCGCCA

[0784] GGCTGTAGAGTTTCCCACCTGCCCTGATCCCTGTCTGAATAGAGATAGCATGA

[0785] CTCAGCGCCGGGGAAAGGGTGGTTCTCTCTGTCTAGCTGATGTCCTCAGCTCC

[0786] AGGAAGTTCTGGGCTTGCGGCATGACAGCAGAGAGAAGGGCCACGCTGCTCA

[0787] CGTCAGCAGTGCCCCACTGAGGGGACCCAATGCAGGAAGGGGCAGCCCAGA

[0788] GGCCAGCCGGCCCCCACCACCAGCTACCGGGTGTGGAGAGGAAACACAGGG

[0789] CAGTGTCTATTTAGATCTTCAGCAAGCTTTCCTT (SEQ ID NO: 88; GLUT7)

[0790] AGGATGCCCAATTGGCTGGGCTGTGCCAGGCATTCCTGAGTTTCCCCA

[0791] TCTCCTCTGGCCACCCCTCACCAGCCCTGGGCCTGGGACCCACCCTCTGGACC

[0792] CCGGCTTCCTTCCTAGGCCCACCTGAGGACTCTGTTCATTTTCCAGGTACCAG

[0793] GGCCAGAGGAGTTGGGGAAGGAAGGAGGGAGGACAGACTGGGCAGCCGGG

[0794] GGTGCCCGGAGCGCTGTTTCTGGGTCACCGGTCTTGGGGAATTCCCCAGTGCC

[0795] CGGAGAGCTGCTTACCTGGTGAGTAACCGCTGCTGGCTGCCCACACAGAATG

[0796] CCTGACTGTGAGGGGACCTCCCGTCCCACCCCTGCCCATGCCACTGCCATGCT

[0797] GCCCTCTCTCCTGAGAGCCCCGGGGTCCAGGAAAGAGACAGACAGGCAGGG

[0798] GGCTGATCCCCACCCCCGCACCCCCCCCCCACCCTGGGACTTTGCAGGGGGC

[0799] GGCTGGTGATTCAAGCCCCTCAGAGCTGAGTTTGT (SEQ ID NO: 89; GLUT7) TCCAAACAGTTCGTACAGTTCTAGGAAACAAGCTGAGTTTGGCCTGTG

[0800] CATTTTGGCATCCTGAACCACAGGATCATGGGGGGGAAGGGGAGAGGAGGG

[0801] AGGATGGAAAAGTACCCGGCAGTGCCTCCTGAATCCAACTTTCTGCTGGCGA

[0802] AATCCACCTCTCGCTGACTCTGAGTCACCAACAGCATGCCTTCCCTCCCCTGC

[0803] GGCCCATCCGCCCATTAGGCCCGGCAGCTCCCCTCCTATGCTGACTTCCCTCG

[0804] CTCGCTCCCACACGTGTGCTCAAGGTGGAGGCAACCCTAATGAGGAAACACG

[0805] AGGCAGCTGCTGTGGCAGGCGTTTCTTCCTTAGCCAGCAAAAGGAGGCCTGC

[0806] AGCAGCCTCGCCTGCAGGACCTGCTGGCCCACTGCTTTGCAGGTCACCCCTCA

[0807] AACCCAGGCTCCCCTCAGCCCTGTAACCTGGAGCTCGCCCCTAGGATGGGGA

[0808] ACCATAGGAATTCTGGGCTGGGCTCATAGACCCC (SEQ ID NO: 90; GLUT7)

[0809] TACTGAGAGAACCGCTAAGAAGGCAGCTATGATGTTTCTTTCGTATTT

[0810] TTTCTCAATGCCTAGCTCTGTGTTTGATGTCTAGTAGGCAACTTATAAAAGCA

[0811] TATTGACAGAATGAATGAGTGGGGCATGATTTTGTATATTCCTTTGCTCAGTC

[0812] CCTTTGCAGTCTATTTTAGATAAACACACATTCTAGTGAGTACCAGGAATTAG

[0813] ACAGCACATATGCATTTCAGAGACTGAGGGTATGAGAAGTCCAGACTCCTCT

[0814] CCCAGCTCTTCTCTCCACTTACAGAGGAATCCAACCTCCTCTCCTTCTCCCCCC

[0815] ACCTCACAGAAGAGGGAGGCCAGCTCTAATGGCCTTGGCCCCAGTTTGCTTG

[0816] TATAATTAATGTGGGATTTTTCCTCCTCTAGTAAAAAATGACCTTGTCAAACA

[0817] AGAGACGACAAAATCTCGGTTACTGAGCGATTGTGGACTCTCTGCTGGGATG

[0818] GCATCCTGGGCCTTGCAAAGAGAAATCTAA (SEQ ID NO: 91; GLUT8)

[0819] GTGACTGGGACTACAGGCACACCAATGCAGCTTAATAGGTTTTCTTTT

[0820] CTACTGTTGCTCTCAAATGCTTAATAGGGGAGGTGGTGTCTTCTCCACAAGAT

[0821] TTTTTTATTCTG(^TAACTTTCTATCTACTTTAATGGTTTCAAAATTAATTTTG

[0822] CTTACCCCACAGAGGCCCTCCTGTGAGTTAATATTAATTGTCAGAAGGGTACA

[0823] TAAATTACATAGTATTGGAGATAGTGTTCTCTATTCAGGAAATCTGATATACA

[0824] AATATGCTGGTAACTCCAAAGTATTTTTTATCAGTCTGATTTGTCTCTTGAGCT

[0825] TCATACTCTCATCTGCCTCCTGGATACGTACCTTCAACTCTGTTGATTGAGCCC

[0826] TCAATTTGTGCCTATTTCTGCATTCACCATTGTCCTGTTGCCCAAACCACCTCC

[0827] TCCCCTCACTCTCATGTTTGGTTGGTTACTTGATTCTCTTCTTTCTGTATTAAA

[0828] AATCTCTTTCCTGTCTCCTCCCA (SEQ ID NO: 92; GLUT8) TTGCAATTGTTATTAAATTCTTTTCCTGGGCTATAATGCATAGGCTTTG

[0829] ATCTAAAGTCCTACCGTTGCTAGAAGGACTCGTATGATTACGATAATGTGCAC

[0830] TGCTCTTTTTGCTATTCCTATAGGGAAAATAAACCCCACATTAGTTCCATATT

[0831] AGTACCTGACTTAGAGAAGTTAAAAAGCACCAGAAAAAGGTTAAACAGAGT

[0832] CAAGTACACATAAGAGAGAAGTGGTTATTAGTGTTCGGGGAGCCATTAAAAA

[0833] ATTCTGCTTATACAGAAGAATTTATTGGTGCTCCTGCTTCTGGCTGCTTTATGC

[0834] ATTTTAATAAAGAAGTAGGCTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGT

[0835] GTGTTGTGTGTGTGCTTGCACACATTTTATGGCTATGATTGAACAAGGACTTT

[0836] AGCTATTTGGTCTGTCATTAATTTTATCCTTTTTTTTTCCTCTCCTTTCCCCAAG

[0837] TCTCCTCCTTTACCTTTTAGCTTCCCA (SEQ ID NO: 93; GLUT8)

[0838] TAGAGAGGCAGTATGTTTACACTTTCTTCCAAAGCAATAGAGAGTCAA

[0839] GTATGCTAGTGAGTAAGTGAATTTTCTATGGAATTTTATTATTCCTCCATATA

[0840] CTTGTTTCATACCAGGTGGAAATCAGGTTTCTAAGAGTAGTTTTATTTCTATG

[0841] CCTTTATAAGAACTTTGTCATTTTCCAAGTAGAATAAATTTGATAAGTAATGA

[0842] TTTATCCTTTTTGTGCTTTTCCTTCTGAACAAGGGCAACTGCTCTCCACTGAAC

[0843] ATTTTACCCTCCCAAGCTGAGTTTGACATAATTACCGCATTGGAGCTAAGATT

[0844] CATAAGCTGATTGCCAATGTCAACGGAAATGAAAGCTTTCATGAAAAACAAA

[0845] ACCCGTTAAGTATACTCAAGGCTCAGCACCAATCCCACTCATGAGTAATTAA

[0846] AATATCAATTTATGATGCATTTGTCTTTTACTGGTAAATCTGGAAATCCTATA

[0847] GATTTTTTTTTTCTTCAGACATAGATGCT (SEQ ID NO: 94; GLUT8)

[0848] AAGTCCTGGGACCCAGGGAGGCCACAGCACTGGAGGACGGCTGCCAG

[0849] CAGGTGGGAAAGGGGCTTGGCCAGGGCGCTGCCTGTCCCGTTCTTTCGTTTGC

[0850] AAGCAGCTCTTGAGTCCACACCTCCAGGGGCCTGCAAGTTCATTTGCTTGTCT

[0851] TACACCTGGCAGCCCTGAGCACTGAGGCTTGATGAGGCTAAGGAGATCTGCC

[0852] AGGAAGACCCTATTGATTTTAATTACCCAGAAACAGCCCACAGGGACAAAGG

[0853] ACTTTGGCTGTTTATTTAGCCAGGCTGTGCTGCGCTATTGTGGAAGGAGAGAT

[0854] CAAATCAGCTGCCCTCATTATAACTATAATAATAAGGCCAAAACACTGCTCG

[0855] GGGCTCAGGGCATTTTTTTTTTTAAAGCAATAACCTCACTGCCCCACCCCCTC

[0856] ACGCCCCTCCTCAGCAGGCTTGAACAACCAGGAGCAATTACATCCTGTCGAT

[0857] ATTAATTCAATTAAATCCAGGTTATTTATTGGG (SEQ ID NO:95; GLUT9) TAAAAGTCAGGAATATGAGCTCCCTGGATTCCACAGCATGTGACTTTA CTGATGGTGGGCCTTACCACATTGCAGGTAATGTGAGCTTCAGAACCTTCTCT CTCTCTGATTAGCCAGCAACGTCAGGGCAAAGCTGAAATCGCTTCATTGTAA CAGTTTGACCTCTGCAATACTGCACATCATTAAATTTTTAAGGGAGTTCATTG GGAAAGTCCTTTAATCCTTTAATAATTGAGTTGGGGGAAAGGGGCCAGTGAT GGAGAGCATTTACCTGAGCTGATATTAACTAGGTTTGCCTACTAATGGCTTTC CAACAATCCCACTGTGTCCATGTTGCCATTTAAAAGAGTGCATTTCATTTGGC CCTTGCTCTTTAACTGTCAACAGGGAAATTGAAAAATAATATAGCATTAAGT ATTGCAATTTGTATTACAGTTCTCTACCCAAGTCTCCCTTCTAGCTCTATTGAG CTCAGTCCAAACTAAGGTCTAACAACCAGT (SEQ ID NO:96; GLUT9)

[0858] GTGACTTCGTTTCCACATCTGTAAATGGGAAAGATCATAATACTATTT ATCTTGTAGGGTCACTGGGAGGATTTAATGACTTAATAATTGTTGAGCACACA TGGAATAGCCTGGAACACATTAAGTACTATAGTAGTATTTGCTACTGTTGTTG TTATTATCAAAGGAGCCTTACAGATATTGGGACATGATGGGACTAGCCATTA TGAGAAGTGCCCTGGGAAATCAATAGGAAATCATTACTGGAAAACACATGAA TGTACAGTGGACATGGCCAGTGTGATCCTGTGTTTATCTTCAGCAAGCAGGG AGCAGAGGCAGAGAGAAGAAAAAGTGGGAATTTCGTCCTGGGGGGATTGGG AAAAAGCAGTGTAGACAGAATGCAAAGTTAGAAGACAGGATTCTAGAGTAG TGATGAGGACATCAAATAGCAAATGTGAAGTTCCAGCCAGGCAAGGAGTCA AATGAAGCTAAGTGTGGACTAATGGACTGATTATGGG (SEQ ID NO: 97; GLUT9)

[0859] Validation'. To validate candidate REs, their genomic positions were visualized. Features that were used to validate included relative levels of open chromatin, potential links to cell population-specific genes, and sufficient amount of open chromatin to drive gene expression. The visualization is shown in FIGS. 11A-11H. Additional details regarding REs having SEQ ID NOS: 1-5 and 21-23 are provided in TABLE 2.

[0860] Example 2 -In vitro and in vivo testing of REs

[0861] P21-28 male and female mice were intraspinally injected with 1 pL of AAV PHP.eb GLUT11.7 GFP (FIG. 3A) at IxlO13GC / mL. Intraspinal injection was carried out as described elsewhere (see, Arokiaraj et al., Contemporary Approaches to the Study of Pain from Molecules to Neural Networks, Vol. 178, Chapter 20, Springer Nature Humana Press, 2022). Three weeks later, mice were perfused with 4% paraformaldehyde, and their spinal cords were dissected out and post-fixed for 12 hours at 4°C. The cords were transferred to 30% sucrose for 3 days and then cut on a cryostat at -20°C into 20 pm sections onto glass slides. The slides were processed for immunohistochemistry (FIG. 3B) with an antibody against Pax2.

[0862] P21-28 male and female mice were then intraspinally injected with 1 pL of the AAV PHP. eb GLUT11.7 hM4Di-GFP vector (FIG. 3C), which was diluted for functional testing to a final concentration of 5x1012GC / mL. Intraspinal injection was carried out as described elsewhere (see, Arokiaraj et al., supra). After three weeks, mice were perfused with 4% paraformaldehyde, and their spinal cords were dissected out and post-fixed for 12 hours at 4°C. The cords were transferred to 30% sucrose for 3 days and then cut on a cryostat at -20°C into 20 pm sections onto glass slides. The slides were processed for immunohistochemistry (FIG. 3D) with an antibody against Pax2, a marker of inhibitory neurons, demonstrating that nearly all GFP+ neurons were Pax2+ and therefore excitatory.

[0863] In further studies, adult mice were injected with AAV PHP. eb GLUT11.7 hM4Di- GFP in the dorsal horn and were subjected to mechanical threshold testing on the plantar hind paw, before and 5 days after intraplantar injection of complete Freund’s adjuvant (CFA) using injection of saline or clozapine-n-oxide (CNO; a synthetic drug that serves as a ligand to activate hM4Di receptor). These studies demonstrated that expression of hM4Di by the GLUT11.7 RE and its subsequent activation by CNO had no effect on baseline mechanical threshold, but caused the mice to recover from the CFA-induced mechanical hypersensitivity (FIG. 3E).

[0864] In another study, P21 C57B1 / 6 male and female mice were injected with AAV8 GLUT11.7 hM4Di T2AEGFP (FIG. 3F) in the dorsal horn. Three weeks later, the mice were perfused and stained with an antibody to Pax2 (FIG. 3G) as described above for FIG. 3C. For functional studies, C57B1 / 6 male and female mice were injected with

[0865] AAV8 GLUT 11.7 hM4Di T2AEGFP in the dorsal horn and were subjected to mechanical threshold testing on the plantar hind paw, before and 9 weeks after intraperitoneal injection of streptozotocin (STZ, 60 mg / kg) for 6 consecutive days to induce a model of diabetic neuropathic pain. Saline or clozapine-n-oxide (CNO; a synthetic drug that serves as a ligand to activate the hM4Di receptor) was injected 20 minutes prior to mechanical testing. These studies demonstrated that expression of hM4Di by the GLUT11.7 RE and its subsequent activation by CNO caused the mice to recover from the STZ-induced mechanical hypersensitivity (FIG. 3H).

[0866] The experimental paradigm used for FIG. 3H was used to test the impact of activating hM4Di on mechanical allodynia that developed 1 and 2 weeks after inducing the spared sural nerve injury model. Saline or CNO was injected 20 minutes prior to mechanical testing. These experiments demonstrated that expression of hM4Di driven by the GLUT11.7 RE and its subsequent activation by CNO caused the mice to recover from the mechanical hypersensitivity induced by this neuropathic injury model (FIG. 31).

[0867] In another study, GLUT5.92 RE (FIGS. 4A-4B) and GLUT6.97 RE (FIGS. 4C- 4D) were subcloned into the AAV plasmid with an intron and with NLS-sfGFP as the cargo, and packaged with the PHP. eb serotype into separate viruses. Each virus was injected individually into adult C57B1 / 6 mice and three weeks later the mice were examined for expression using a rabbit anti-GFP antibody (FIGS. 4A-4D). These studies demonstrated expression of sfEGFP by the GLUT5.92 and GLUT6.97 REs in the dorsal horn of the mouse spinal cord.

[0868] Attorney Docket No. 26100-0147W01 CMU 2023-091 / Pitt 06158

[0869] TABLE 2

[0870] OTHER EMBODIMENTS

[0871] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

WHAT IS CLAIMED IS:

1. A nucleic acid construct comprising:(a) a nucleotide sequence encoding a polypeptide, and(b) at least one regulatory element (RE) specific for a selected cell population, wherein the RE comprises the nucleotide sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, or SEQ ID NO:23, or comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, or SEQ ID NO:23, wherein the RE is operably linked to the sequence encoding the polypeptide and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population.

2. The nucleic acid construct of claim 1 , wherein the polypeptide is a Designer Receptor Exclusively Activated by Designer Drug (DREADD) polypeptide.

3. The nucleic acid construct of claim 2, wherein the DREADD polypeptide is a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4- 5HT3 polypeptide.

4. The nucleic acid construct of claim 1 , wherein the polypeptide is a polypeptide that reduces transmitter release from neurons.

5. The nucleic acid construct of any one of claims 1 to 4, wherein the nucleic acid further comprises a nucleotide sequence encoding a tag polypeptide, such that when the nucleotide sequences encoding the polypeptide and the tag polypeptide are expressed, the polypeptide is coupled to the tag polypeptide.

6. The nucleic acid construct of claim 5, wherein the tag polypeptide is a fluorescent polypeptide.

7. The nucleic acid construct of claim 6, wherein the fluorescent polypeptide is a green fluorescent protein (GFP).

8. The nucleic acid construct of claim 7, wherein GFP comprises an amino acid sequence having at least 95% sequence identity with the superfolder GFP sequence set forth in SEQ ID NO:20.

9. The nucleic acid construct of claim 6, wherein the fluorescent polypeptide is selected from the group consisting of mCherry, mCitrine, mRuby, nuclear localization sequence-GFP, tdTomato, and SunlGFP10. The nucleic acid construct of any one of claims 1 to 9, wherein the construct further comprises virus sequences.

11. The nucleic acid construct of claim 10, wherein the virus sequences are adeno- associated virus (AAV) sequences or lentivirus sequences.

12. The nucleic acid construct of any one of claims 1 to 11, wherein the RE is specific for a population of dorsal horn spinal cord neurons, such that the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons transduced with the nucleic acid construct but is not expressed in at least 90% of other dorsal horn spinal cord neuron populations transduced with the nucleic acid construct.

13. A virus particle comprising the nucleic acid construct of any one of claims 1 to 12.

14. The virus particle of claim 13, wherein the virus is AAV or lentivirus.

15. A method for modifying a selected cell population within a broader population of cells, wherein the method comprises introducing into the broader population of cells a nucleic acid construct, wherein the nucleic acid construct comprises:(a) a sequence encoding a polypeptide, and(b) at least one RE specific for the selected cell population, wherein the RE comprises the nucleotide sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, or SEQ ID NO:23, or comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO: 22, or SEQ ID NO: 23, wherein the RE is operably linked to the sequence encoding the polypeptide, and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population, and wherein the polypeptide is expressed in the selected cell population.

16. The method of claim 15, wherein the polypeptide is a Designer Receptor Exclusively Activated by Designer Drug (DREADD) polypeptide.

17. The method of claim 16, wherein the polypeptide is a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4-5HT3 polypeptide.

18. The method of claim 15, wherein the polypeptide is a polypeptide that reduces transmitter release from neurons.

19. The method of any one of claims 15 to 18, wherein the nucleic acid further comprises a nucleotide sequence encoding a tag polypeptide, such that when the nucleotide sequences encoding the polypeptide and the tag polypeptide are expressed, the polypeptide is coupled to a tag polypeptide.

20. The method of claim 19, wherein the tag polypeptide is a fluorescent polypeptide.

21. The method of claim 20, wherein the fluorescent polypeptide is a GFP.

22. The method of claim 21, wherein the GFP comprises an amino acid sequence having at least 95% sequence identity with the sequence set forth in SEQ ID NO:20.

23. The method of claim 20 wherein the fluorescent polypeptide is selected from the group consisting of mCherry, mCitrine, mRuby, nuclear localization signal-GFP, tdTomato, and SunlGFP.

24. The method of any one of claims 15 to 23, wherein the nucleic acid construct further comprises virus sequences.

25. The method of claim 24, wherein the virus sequences are AAV sequences or lentivirus sequences.

26. The method of any one of claims 15 to 25, wherein the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced, but is not expressed in at least 90% of dorsal horn spinal cord neurons of other populations into which the nucleic acid construct was introduced.

27. A method for treating pain in a mammal, wherein the method comprises administering, to the mammal, a nucleic acid construct comprising:(a) a sequence encoding a polypeptide, and(b) at least one RE specific for a selected cell population, wherein the RE comprises the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, or SEQ ID NO:23, or comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO: 22, or SEQ ID NO: 23, wherein the RE is operably linked to the sequence encoding the polypeptide, and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population, and wherein the polypeptide is expressed in the selected cell population within the mammal.

28. The method of claim 27, wherein the mammal is a human.

29. The method of claim 27 or claim 28, wherein the polypeptide is a Designer Receptor Exclusively Activated by Designer Drug (DREADD) polypeptide.

30. The method of claim 29, wherein the polypeptide is a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4-5HT3 polypeptide.

31. The method of claim 27 or claim 28, wherein the polypeptide is a polypeptide that reduces transmitter release from neurons.

32. The method of any one of claims 27 to 31, wherein the nucleic acid construct further comprises virus sequences.

33. The method of claim 32, wherein the virus sequences are AAV sequences or lentivirus sequences.

34. The method of any one of claims 27 to 33, wherein the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons into which the nucleic acid construct was introduced, but is not expressed in at least 90% of dorsal horn spinal cord neurons of other populations into which the nucleic acid construct was introduced.

35. A method for treating pain in a mammal, wherein the method comprises administering, to the mammal, a virus comprising a nucleic acid that comprises:(a) a sequence encoding a polypeptide, and(b) at least one RE specific for a selected cell population, wherein the RE comprises the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO:22, or SEQ ID NO:23, or comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:21, SEQ ID NO: 22, or SEQ ID NO: 23,wherein the RE is operably linked to the sequence encoding the polypeptide, and is effective to drive expression of the sequence encoding the polypeptide in the selected cell population, and wherein the polypeptide is expressed in the selected cell population within the mammal.

36. The method of claim 35, wherein the mammal is a human.

37. The method of claim 35 or claim 36, wherein the polypeptide is a Designer Receptor Exclusively Activated by Designer Drug (DREADD) polypeptide.

38. The method of claim 37, wherein the polypeptide is a hM4Di polypeptide, a hM3Dq polypeptide, a PSAM4-GlyR polypeptide, or a PSAM4-5HT3 polypeptide.

39. The method of claim 35 or claim 36, wherein the polypeptide is a polypeptide that reduces transmitter release from neurons.

40. The method of any one of claims 35 to 39, wherein the virus is an AAV or a lentivirus.

41. The method of any one of claims 35 to 40, wherein the sequence encoding the polypeptide is expressed in a majority of the population of dorsal horn spinal cord neurons to which the virus was administered, but is not expressed in at least 90% of dorsal horn spinal cord neurons of other populations to which the virus was administered.IQ