method

Abstract

This disclosure relates to a chemogenetic method that enables the use of a drug at low doses by overexpressing the target of the drug to be administered, and to a vector for introducing a nucleotide sequence into the target cells for the purpose of enabling such overexpression.

Description

【Technical Field】 【0001】 The present invention relates to a method for reducing the side effects of a neuromodulatory agent in order to enhance the effectiveness of the neuromodulatory agent, and a method for reducing the excitability of neurons. The present invention also relates to a gene therapy vector useful for carrying out the method of the present invention. 【Background Art】 【0002】 Neuropathy is characterized by abnormal activity of neurons and circuits within the central nervous system (CNS). This abnormal activity is often transmitted to the output neurons of the CNS, i.e., motor neurons, causing inappropriate muscle contractions and sensory and motor dysfunctions. 【0003】 Typically, drug therapies have been developed aimed at improving abnormal activity by regulating ion channels that carry excitatory and inhibitory currents. However, drug delivery to the nervous system is difficult, and existing therapies often have undesirable side effects due to low specificity. 【0004】 Thus, although some neuromodulatory agents developed for neuropathy have shown some therapeutic advantages, there have been limitations; the reason being that they have off-target effects on non-dysfunctional neurons and other systems that express the target (i.e., cardiovascular, respiratory, gastrointestinal, etc.). 【0005】 Potassium (K + ) channels are membrane proteins that allow for rapid and selective flow of K + ions across the cell membrane, and thus generate electrical signals in cells. Voltage-dependent K +Kv channels open and close in response to changes in membrane potential. Kv channels are one of the main components of the generation and propagation of electrical stimuli in the nervous system. Changes in membrane potential cause these channels to open, allowing passive ion flow from the cell and restoring the resting membrane potential. Therefore, activation of Kv channels is considered inhibitory to action potential generation. 【0006】 There are 40 human voltage-gated potassium channel genes belonging to 12 subfamilies (Kv1-Kv12). These subfamilies are typically limited to specific sites or cell / tissue types, and their presence is found in both excitable and non-excitable tissues. Mixing and combining Kv channel subunits can lead to a high degree of diversity in Kv channels. Within each of the Kv1, Kv2, Kv3, Kv4, and Kv7 families, homozygous and heterozygous channels can be formed, resulting in a wide range of functional characteristics. Kv2 family members can also complex with Kv5, Kv6, Kv8, or Kv9 family members, which have more limited expression patterns in the nervous system and smooth muscle. 【0007】 Voltage-gated potassium channels in neurons target various intracellular compartments, and channels with different subunit compositions may exist in different subpopulations of neurons. 【0008】 The tetrameric structure of the Kv channel consists of two functionally and structurally independent domains: an ion permeation pore and a voltage-sensing domain. The ion permeation pore consists of four subunits symmetrically arranged around the conduction pathway. The voltage-sensing domain consists of four transmembrane segments (S1-S4) located around the channel. Structural reconfiguration of the voltage-sensing domain in response to changes in membrane potential, particularly S4 (containing positively charged amino acids at residue positions every three residues), leads to structural changes in the permeation pore (opening or closing the ion conduction pathway). The voltage-dependent mechanism is not yet fully understood. 【0009】 Low threshold voltage-dependent potassium (K) + The Kv7 family of channels consists of five members (Kv7.1–7.5), each encoded by the KCNQ1–5 genes. Kv7 channels are constructed as tetramers of identical or interchangeable α-subunits, each consisting of six transmembrane segments (S1–S6) as well as cytoplasmic N-terminal and C-terminal segments. Kv7 is widely expressed in both the CNS and other tissues, and Kv7.2 and Kv7.3, in particular, are known to regulate neuronal excitability. 【0010】 Kv7.2 and Kv7.3 channels typically form heteromers with each other, increasing conductivity, but are rarely expressed as low-conductivity homomers. Kv7.2 and Kv7.3 are expressed in most neuronal subtypes and tend to localize to the axonal origin where action potentials (spikes) initiate. Therefore, the conductivity of Kv7.3 and Kv7.2 is considered a particularly potent inhibitor of action potential generation and activity in the same neuron and its targets (e.g., inhibiting motor neurons reduces muscle activity). 【0011】 Neuromodulatory agents targeting Kv7 channels have been developed to treat disorders associated with neuronal hyperexcitability. Retigavin is an anticonvulsant that primarily acts as a Kv7.2-Kv7.5 potassium channel opener, and at higher concentrations, acts as a GABA-positive allosteric regulator. The term "channel opener" refers to a leftward shift in the voltage dependence of channel opening toward more negative potentials. In the presence of retigabine, Kv7 channels open at more negative potentials, contributing to membrane hyperpolarization and voltage stabilization. 【0012】 Retigabin has also been shown to stabilize the opening state of Kv7.2 / 7.3 (heterogeneous complex) channels and to slow down inactivation with little change to the voltage dependence of inactivation. The overall effect is increased potassium conductivity, hyperpolarization of membrane potential, and suppression of action potential generation. This effect of retigabine has been observed in vitro at concentrations of less than 10 μM (Corbin-Leftwich et al., 2016; Villalba-Galea, 2020). Oral administration of retigabine at 600-1200 mg / day (Gunthorpe et al., 2012), corresponding to an average plasma concentration of 0.83 μM, has been shown to have up to 45% clinical efficacy in reducing seizures in people with epilepsy (Brodie et al., 2010; French et al., 2011). In particular, the EC50 of retigabine is 0.6 μM for the Kv7.3 channel, while most channels in the nervous system are Kv7.2 / 7.3 heteromers, and their EC50 is 1.6 μM. In some patients, these doses caused discoloration of the eyes and skin, which led to limitations in its indications. Subsequently, retigabine was voluntarily withdrawn for commercial reasons (Brickel et al., 2020). 【0013】 There remains pharmaceutical interest in Kv7 as a therapeutic target. Clinical trials have been conducted on at least three activators that exhibit higher efficacy, greater selectivity for Kv7.2 / 7.3, and no cosmetic side effects, showing promising results. However, because Kv7.2 / Kv7.3 expression is ubiquitous in the nervous system, neurological side effects (drowsiness, fatigue, etc.) are currently unavoidable. Although novel Kv7.2 / 7.3 activators have improved safety profiles, neurological side effects can only be mitigated by enhancing selectivity to dysfunctional neurons only. 【0014】 Chemogenetics is a method of engineering proteins to interact with small chemical actuators that the protein was previously unable to recognize. Since the 1990s, numerous chemogenetic platforms have been developed, proving particularly useful for neuroscientists attempting to modulate the neural activity of specific populations. Several classes of proteins have been engineered using this method, including kinases, non-kinase enzymes, ligand-gated ion channels, and G protein-coupled receptors (GPCRs). Among these, the most widely used are designer receptors activated by designer drugs (DREADDs). 【0015】 DREADD is an engineered GPCR that does not respond to its natural ligand, acetylcholine, but does respond to synthetic ligands that cross the blood-brain barrier (such as clozapine-N-oxide and olanzapine). 【0016】 For use, a viral vector is used to introduce the gene encoding the DREADD protein into the target cells. Various viral serotypes, promoters, and administration routes can be used to assist in the selection of target cells. Once gene transfer is complete, it takes 2-3 weeks for the infected cells to express the engineered receptor protein, after which it can be activated with the DREADD ligand. In this way, the activity of target neurons can be specifically regulated while minimizing the impact on non-target cells. 【0017】 The method described herein is similar to that used in DREADD, but involves expressing drug-responsive ion channels in neurons to modulate their activity. Importantly, this method overturns the existing concept of designing specifically paired de novo receptors and drugs, and repurposes wild-type and mutant ion channels for the purpose of mitigating or eliminating the side effects of drugs that act on the channels. This method involves overexpressing wild-type and / or mutant channels. 【0018】 Furthermore, the method described has the advantage of utilizing a channel that can be opened (agonist) or closed (antagonist) depending on the drug used. In contrast, activation of DREADD only induces a unidirectional excitatory change (either excitatory or inhibitory). [Overview of the project] 【0019】 This disclosure relates to a chemogenetic method that enables the modulation of the activity of targeted neurons, thereby increasing drug efficacy while reducing side effects based on the possibility of using lower doses, at least in part. The method involves using a gene therapy methodology via an introduction vector (e.g., AAV) to overexpress (upregulate) a therapeutic ion channel in a target neuron, wherein the conduction of the channel (and therefore the activity of the neuron) can be altered by low doses of specific activators and inhibitors. In other words, the method specifically increases the sensitivity of dysfunctional neurons to modulatory agents. 【0020】 The inventors demonstrated that AAV-mediated overexpression of the voltage-gated potassium channel Kv7.3 (encoded in KCNQ3) allows Kv7 activators (such as retigabine) to significantly reduce the excitability of human neurons at both the individual neuron and population levels. At low concentrations (0.3–3 μM), retigabine reduced the number of action potentials in individual hIPSC-derived sensory and motor neurons. At the same dose, retigabine had minimal effect on control neurons expressing only green fluorescent protein. Using a multi-electrode array, the inventors showed that retigabine is effective in selectively reducing spontaneous burst activity from networks of thousands of MNs overexpressing Kv7.3, but does not have such an effect on GFP-expressing neurons. These in vitro results were applied to a mouse spasticity model, in which overactive MNs cause excessive muscle contractions (muscle spasms). In this model, intramuscular injection of the AAV-Kv7 construct resulted in motor neurons overexpressing Kv7.3, allowing for a reduction in the intensity and duration of muscle spasms with low-dose retigabine (2 mg / kg). However, mice injected with the AAV-GFP construct did not show a significant reduction in spasticity at the same dose. 3 μM of retigabine reduced AP firing in mouse motor neurons expressing Kv7.3, but this was not the case for GFP-expressing neurons; therefore, reduced MN excitability was confirmed as a mechanism for reducing spasticity. Furthermore, we demonstrated that introducing a point mutation at amino acid residue 315 of the Kv7.3 protein reduced baseline motor neuron excitability both in vitro and in vivo, and further increased the effect size of retigabine at the concentrations tested in vitro (Zaika et al., 2008; Gomez-Posada et al., 2010). 【0021】 In this therapy, AAV is used to overexpress the Kv7.3 ion channel subunit and its variations as either homomers or Kv7.2 / 7.3 heteromers in the hyperexcitable neurons of patients suffering from neuropathy, and a low dose of channel modulator is used to normalize the activity. 【0022】 According to a first aspect, there is provided a method for enhancing the effectiveness of a neuromodulatory agent; The method comprises: (a) introducing, via a vector, a nucleotide sequence encoding a Kv7.3 ion channel subunit comprising any one of the sequences of SEQ ID NO: 1 to SEQ ID NO: 8; or a nucleotide sequence encoding an amino acid sequence of a Kv7.3 ion channel subunit of any one of SEQ ID NO: 9 to SEQ ID NO: 16, into target neurons; (b) overexpressing the ion channel subunit in the target neurons to form functional ion channels; and (c) administering the neuromodulatory agent at a lower dose, wherein the method includes the above steps. 【0023】 Advantageously, by enhancing the effectiveness of the agent, a lower dose can be administered to achieve the same therapeutic effect. This means, beneficially, that the occurrence of side effects can be reduced. 【0024】 Thus, in a second aspect, there is provided a method for reducing the side effects of the neuromodulatory agent; The method comprises: (a) introducing, via a vector, a nucleotide sequence encoding a Kv7.3 ion channel subunit comprising any one of the sequences of SEQ ID NO: 1 to SEQ ID NO: 8; or a nucleotide sequence encoding an amino acid sequence of a Kv7.3 ion channel subunit of any one of SEQ ID NO: 9 to SEQ ID NO: 16, into target neurons; (b) Overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) Administering the neuromodulatory agent at a lower dose, which includes. 【0025】 The side effects of neuromodulatory agents can range from mild to severe; they can also affect the quality of life of the subject. For example, regarding retigabine, the NIH has listed the most common adverse effects; adverse effects occurring in more than 10% of subjects in clinical trials include abnormal gait, confusion, dizziness, fatigue, headache, nausea, drowsiness, speech disorder, tremor, urinary tract infection, and blurred vision (Harris and Murphy, 2011). 【0026】 In some cases, the required dose causes unacceptable side effects (adverse effects), so it is possible that the drug is removed from the market due to such adverse effects. If the dose can be reduced, the adverse effects can be reduced, and potentially the use of the drug can be resumed. 【0027】 In a third aspect, a method for increasing the therapeutic index of a neuromodulatory agent is provided; The method is: (a) A nucleotide sequence encoding a Kv7.3 ion channel subunit comprising any one of the sequences of SEQ ID NO: 1 to SEQ ID NO: 8 via a vector; or A nucleotide sequence encoding an amino acid sequence of any one of the Kv7.3 ion channel subunits of SEQ ID NO: 9 to SEQ ID NO: 16, introducing it into the target neuron; (b) Overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) Administering the neuromodulatory agent, which includes. 【0028】 In the fourth phase: A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, We provide gene therapy vectors that include; When this vector is used to transduce target neurons, the expression of the Kv7.3 ion channel subunit is upregulated, and a functional ion channel is formed. 【0029】 Advantageously, introducing the nucleotide sequence into a specific population of target neurons results in the nucleotide payload being delivered only to neurons exhibiting increased excitability. These neurons then overexpress the ion channels, presenting more targets for the neuromodulatory agent; this allows the agent to be effective at lower doses, thus reducing side effects. Targeting neurons in this manner also means a reduced possibility of off-target activity. 【0030】 In the fifth phase: (a) AAV capsids that selectively induce target neurons; and (b) A nucleotide sequence encoding a Kv7.3 ion channel subunit containing any sequence from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The invention provides a gene therapy vector for AAV virus particles containing the virus; When this vector is used to transduce target neurons, the expression of the Kv7.3 ion channel subunit is upregulated, and a functional ion channel is formed. 【0031】 In another aspect: AAV capsids that selectively induce plasma delivery to target neurons; Furthermore A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector containing; A gene therapy vector used to upregulate the expression of the Kv7.3 ion channel subunit by the target neuron, thereby enabling the formation of a functional ion channel in a subject suffering from a neurological disorder associated with increased neuronal excitability. To provide. 【0032】 In another scenario: AAV capsids that selectively induce plasma delivery to target neurons; Furthermore A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector for use in the treatment of subjects with neurological disorders, including; Herein, when the vector is transduced into a target neuron, the expression of the Kv7.3 ion channel by the target neuron is upregulated, thereby enabling effective administration of a neuromodulatory drug to the target at a lower dose. To provide. 【0033】 In another scenario: A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, Applications of gene therapy vectors including; Uses in the preparation of therapeutic drugs used to treat neurological disorders associated with increased neuronal excitability, It provides; Here, the neurological disorder is: Epilepsy, spasticity, BNFS, Parkinson's disease, nociceptive and non-nociceptive pain, ALS, Selected from the group consisting of 【0034】 For the purpose of providing a better understanding of the present invention and demonstrating feasible methods of its implementation, specific embodiments, methods, and processes according to the present invention are described below, with reference to the accompanying drawings, for illustrative purposes only. [Brief explanation of the drawing] 【0035】 [Figure 1] The diagram shows an AAV construct used for transduction into neurons, containing either Kv7.3 (construct shown at the top) or Kv7.3 A315T (construct shown at the bottom). Note that the GFP sequence is linked downstream of Kv7.3 and Kv7.3 A315T via a T2A linker. This means that the GFP fluorescence indicates the expression of Kv7.3. [Figure 2]This shows that AAV transduction of hIPSC neurons induces the expression of proteins encoded by the transgene incorporated into the AAV construct (A). Specifically, in this example, AAV was used to transduce hIPSC neurons with the aim of inducing the expression of either green fluorescent protein (AAV-GFP) or human Kv7.3 channel and GFP (AAV-Kv7.3). Figure 2 also shows baseline and post-exposure endpoints (Kv7.3 or GFP expression in the absence of the activator, B) evaluated using patch-clamp electrophysiology. These endpoints are: base current (minimum input current to elicit one action potential); number of action potentials in 3x base current; and resting membrane potential. In the presence of retigabine, neuronal excitability decreased, as evidenced by an increase in base current, a decrease in the number of action potentials in 3x base current, and hyperpolarization of the resting membrane potential. [Figure 3] In comparison with control neurons transduced with AAV-GFP, baseline overexpression of Kv7.3 does not significantly alter the excitability of hIPSC sensory neurons. Panel (A) shows the mean number of action potentials generated at each input current value for AAV-GFP (dashed line) and AAV-Kv7.3 (solid line) transduced neurons. Shaded areas indicate 95% confidence intervals. Excitability levels were judged to be similar when the measured effect sizes for AP / 500 ms (B), RMP (C), and baseline current (D) were extremely small. In these Cummings estimate plots, individual points represent the number of action potentials generated at 3x baseline current for neurons transduced with AAV-GFP (light gray) or AAV-Kv7.3 (dark gray). To the right of the plots, Hedges' G effect size and 95% confidence intervals for bootstrap data resampled (1000 repeats) are plotted. [Figure 4]The results, as determined by measuring the number of action potentials in 3x current, show that retigabine delivered at concentrations of 0.3 μM, 1 μM, and 3 μM had only a limited effect on the excitability of AAV-GFP transduced hIPSC sensory neurons. However, when measured at the same concentrations in AAV-Kv7.3 transduced neurons, large and very large effect sizes were obtained. In each of the estimated plots (A, B, and C), the upper graph is a line plot showing the change in AP count between measurement records taken before and after exposure to retigabine. The lower plot shows Hedges' G effect size and confidence interval. [Figure 5] When determined by measuring the baseline current (the minimum current required to generate one action potential), retigabine delivered at concentrations of 0.3 μM, 1 μM, and 3 μM showed only limited effects on AAV-GFP transduced hIPSC sensory neurons. However, when measured at the same concentrations in AAV-Kv7.3 transduced neurons, a large effect size was obtained. [Figure 6] Measurements of resting membrane potential (RMP) indicated that retigabine delivered at concentrations of 0.3 μM, 1 μM, and 3 μM exerted only small to moderate effects on the excitability of AAV-GFP-transduced hIPSC sensory neurons. However, when measured at the same concentrations in neurons transduced with AAV-Kv7.3, large and very large effect sizes were observed. [Figure 7] Baseline expression of Kv7.3A315T (dashed line) significantly reduces the excitability of hIPSC sensory neurons. AAV-Kv7.3A315T transduced neurons were shown to have reduced excitability levels because they could not generate potentials exceeding three action potentials (A, B). RMP was also hyperpolarized at baseline (C), but the baseline current remained virtually unchanged (D). [Figure 8]This study shows that retigabine delivered at concentrations of 0.3 μM, 1 μM, and 3 μM resulted in a very large increase in basal current in AAV-Kv7.3A315T transduced neurons. Compared to AAV-GFP, the effect size was at least 2–3 times larger in AAV-Kv7.3A315T. Note the differences in scales for both basal current (first axis) and corresponding Hedges' g effect size (second axis). [Figure 9] The results show that retigabine delivered at concentrations of 0.3 μM, 1 μM, and 3 μM had a very significant effect on RMP hyperpolarization in AAV-Kv7.3A315T transduced hIPSC sensory neurons. The effect size was also at least 2–3 times larger than that in AAV-GFP transduced hIPSC neurons. [Figure 10] Western blot (A) and ELISA (B) show overexpression of Kv7.3 in hIPSC motor neurons. Note the dense band around approximately 80 kDa. [Figure 11] Overexpression of Kv7.3 in hIPSC-derived motor neurons resulted in a slight decrease in the baseline frequency / current relationship (A-B), but did not affect action potential firing in 3x basal current (C) and basal current (D). (E-H) Overexpression of Kv7.3 increased the sensitivity of hIPSC MNs to retigabine. At concentrations below 3 μM, it did not affect the excitability of AAV-GFP transdextrins, but reduced AP firing in AAV-Kv7.3 transdextrins by up to 80%. Overexpression of Kv7.3 also enhanced the efficacy of retigabine, as indicated by the increased maximum effect on AP frequency (G) and FI slope (H). [Figure 12]Measurements using a multi-electrode array (MEA) plate demonstrate that overexpression of Kv7.3 increases the susceptibility of MNs to RTG at a population level. hIPSC MNs were grown in well plates with electrodes imprinted on the substrate to allow recording of spontaneous activity. 50% of the wells were treated with AAV-GFP, and the remaining 50% with AAV-Kv7.3. After sufficient time for transduction, spontaneous activity at baseline (0 μM) was recorded, and then the RTG concentration was increased. RTG did not affect the viability of motor neurons as assessed by electrode electrical resistance (A). However, MNs treated with RTG showed greater susceptibility to RTG, as evidenced by a leftward shift in the dose response to retigabine and by the spontaneous action potential burst frequency (B), burst duration (C), or total number of spikes. Sections E-F show representative examples of activity recorded after the addition of 1.8 μM (RTG) in wells treated with AAV-GFP(E) or AAV-Kv7.3(F). The vertical lines in the upper panel indicate network burst frequencies. The lower panel is a raster plot of action potential firings recorded at each of the 16 electrodes (rows). Sections G-H show representative examples of individual bursts from MN in wells treated with AAV-GFP(G) or AAV-Kv7.3(H). Each row represents spike activity recorded at a single electrode. To the right of each plot is a heat legend showing the change in firing frequency. [Figure 13]This study demonstrates that Kv7.3 can be well overexpressed in motor neurons in a neonatal mouse model of spasticity induced by spinal cord injury after intramuscular injection of AAV-Kv7.3. (A) shows Western blots for Kv7.3 in spinal cord samples collected 2 weeks after injection and spinal cord injury. Note the dense band at the expected molecular weight position for Kv7.3 in spinal cord samples from AAV-Kv7.3-treated mice; this band is not observed in AAV-GFP-treated mice. (B) shows immunohistochemistry showing specific Kv7.3 expression in MN. (C-D) shows changes in muscle spasm duration after administration of 2 mg / kg RTG in AAV-GFP-treated and AAV-Kv7.3-treated mice. E-F show changes in spasm intensity (number of action potentials per spasm) after administration of 2 mg / kg RTG in AAV-GFP-treated and AAV-Kv7.3-treated mice. G to H represent typical electromyography (EMG) outputs showing the effect of 2 mg / kg of RTG in AAV-GFP treated mice and AAV-Kv7.3 treated mice. The maximum output is pre-RTG, and the lower output is 20 minutes after RTG. [Figure 14]This study demonstrates that Kv7.3 overexpression in motor neurons via intramuscular injection of AAV-Kv7.3 does not significantly affect the baseline electrophysiological properties of motor neurons in a neonatal mouse spasticity model. The following are records of measurements performed on spinal cord sections taken from spastic mice. (A) shows that when there was no difference in the number of action potentials induced by a 2x current between AAV-GFP control transducer MNs and AAV-SRx-C490 transducer MNs, the excitability of the MNs did not decrease. (B) shows that the threshold for eliciting a single action potential did not differ in mouse AAV-SRx-C490 transducer MNs. (C) shows that there was no difference in resting membrane potential (RMP) between the groups. (D) shows that there was no difference in MN size between the groups. (E~H) indicates that, as evaluated by changes in the number of action potentials at 2x threshold and baseline current, excitability was significantly reduced in MN mice treated with AAV-SRxC490 by 3 μM retigabine, but no such reduction was observed in AAV-GFP treated mice. [Figure 15] Motor neurons in spastic mice treated with AAV-GFP fired as repeating sequences of action potentials (A), but motor neurons in mice introduced with a viral construct to overexpress mutant Kv7 (A315T) were unable to fire repeatedly. This suggests a permanent and significant decrease in the excitability of these motor neurons. [Modes for carrying out the invention] 【0036】 As used herein, “enhancing efficacy” means increasing the likelihood of obtaining the desired outcome. This may be achieved by improving the response to the drug rather than by improving the efficacy of the drug. Improved efficacy means that the same level of response can be achieved with a smaller amount of the drug. 【0037】 Therefore, increasing effectiveness means that lower doses can be used. 【0038】 As used herein, “neuromodulatory agents” refers to voltage-gated potassium channel targeting agents. Known agents include, but are not limited to, retigabine and its derivatives (see, e.g., Musella et al., 2022), BHV-7000 and Xen496, Xen1101, flupirtin, diclofenac, BMS-204352, meclofenamic acid, ETX-123, and linopyrdin. 【0039】 In one embodiment, the neuromodulatory agent is a channel opener. 【0040】 In one embodiment, the neuromodulatory agent is a channel blocker. 【0041】 In one embodiment, the neuromodulatory agent is selected from retigabine or its derivatives, BHV-7000 and Xen496, Xen1101, flupirtin, diclofenac, BMS-204352, meclofenamic acid, ETX-123, and linopyrdin. 【0042】 In this specification, "introducing a nucleotide sequence" refers to transduction or transformation of cells (such as neurons) using one or more nucleotide sequences, for the purpose of supplying existing components to an existing genetic material or providing a novel nucleotide sequence. 【0043】 The Kv7.3 ion channel subunit is a monomer of a voltage-gated potassium channel, which associates to form a tetrameric functional ion channel. Kv7.3 can associate as a homomer or as a heteromer with other Kv7 subunits, including Kv7.2 and Kv7.5, or other auxiliary subunits (e.g., KCNE). 【0044】 In this specification, "Kv7.3 ion channel" is intended to refer to both the Kv7.3 homomer and the Kv7.3 heteromer. 【0045】 In this specification, "nucleotide sequence encoding a Kv7.3 ion channel subunit" refers to a nucleotide sequence (RNA or DNA) encoding a Kv7.3 ion channel subunit. 【0046】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 1 or a variant of SEQ ID NO: 1 that encodes the same amino acid sequence as encoded by SEQ ID NO: 1. 【0047】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 2 or a variant of SEQ ID NO: 2 that encodes the same amino acid sequence as encoded by SEQ ID NO: 2. 【0048】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 3 or a variant of SEQ ID NO: 3 that encodes the same amino acid sequence as encoded by SEQ ID NO: 3. 【0049】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 4 or a variant of SEQ ID NO: 4 that encodes the same amino acid sequence as encoded by SEQ ID NO: 4. 【0050】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 5 or a variant of SEQ ID NO: 5 that encodes the same amino acid sequence as encoded by SEQ ID NO: 5. 【0051】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 6 or a variant of SEQ ID NO: 6 that encodes the same amino acid sequence as encoded by SEQ ID NO: 6. 【0052】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 7 or a variant of SEQ ID NO: 7 that encodes the same amino acid sequence as encoded by SEQ ID NO: 7. 【0053】 In one embodiment, the nucleotide sequence includes the sequence of SEQ ID NO: 8 or a variant of SEQ ID NO: 8 that encodes the same amino acid sequence as encoded by SEQ ID NO: 8. 【0054】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to SEQ ID NO: 9. 【0055】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to SEQ ID NO: 10. 【0056】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to SEQ ID NO: 11. 【0057】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to SEQ ID NO: 12. 【0058】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to SEQ ID NO: 13. 【0059】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to SEQ ID NO: 14. 【0060】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to Sequence ID No. 15. 【0061】 In one embodiment, the Kv7.3 ion channel subunit includes or consists of an amino acid sequence according to SEQ ID NO: 16. 【0062】 In one embodiment, the nucleotide sequence encodes a Kv7.3 ion channel subunit that follows any one of sequence numbers 9 to 16. 【0063】 In certain embodiments, the Kv7.3 ion channel subunit may contain a mutation at the position of amino acid residue 315 of SEQ ID NO: 9. For example, A315T, A315S, A315V, A315C, A315N, A315Q, or A315Y. 【0064】 In one embodiment, the ion channel subunit is wild-type Kv7.3. 【0065】 In one embodiment, the ion channel subunit is not wild-type Kv7.3. 【0066】 As used herein, "functional ion channel" means that the ion channel subunits associate to form a tetramer, which is either a homomer or a heteromer, and is localized on the neuronal membrane at a site suitable for acting as a voltage-gated potassium channel. 【0067】 As used herein, the term “neuron” includes a neuron and its parts (singular or plural) (e.g., the neuron cell body, axon, and / or dendrites). The term “neuron” refers to a cell of the nervous system that is electrically active and includes a cell body (or soma) and up to two types of extensions or processes, namely dendrites and axons (by which the majority of neuronal signals are transmitted to the cell body; and by which the majority of neuronal signals are transmitted from the cell body to effector cells (such as target neurons or muscles)). Neurons are afferent or sensory neurons that can transmit information from tissues and organs to the central nervous system; and also able to transmit signals from the central nervous system to effector cells (efferent or motor neurons). Other neurons, called interneurons, connect neurons within the central nervous system (brain and spinal cord). Yet another type of neuron, called “projection” neuron, extends its axon from one region of the nervous system to another. 【0068】 In some embodiments, the neuron is involved in sensory / motor activity. 【0069】 In some embodiments, the neuron is a motor neuron. 【0070】 In some embodiments, the neuron is a sensory neuron. 【0071】 In some embodiments, the neuron is an autonomic efferent neuron. 【0072】 In some embodiments, the neuron is an autonomic sensory neuron. 【0073】 In some embodiments, the neuron is a subthalamic nucleus neuron. 【0074】 As used herein, “vector” refers to any suitable gene transfer vector known in the art. This includes, but is not limited to, viral vectors (including adeno-associated viruses, adenoviruses, and lentiviruses) and non-viral methods (such as naked DNA or chemically assisted delivery methods). Typically, such vectors are gene therapy vectors. 【0075】 As used herein, “overexpression” is synonymous with upregulation of a gene and is intended to refer to increased gene expression compared to cells (neurons) that have not undergone transformation or transduction. For this to be effective, the neuron must express the complexed Kv7.3-containing ion channel at a higher level, either as a homomer or heteromer (e.g., Kv7.2 / 7.3 heteromer) of Kv7.3. 【0076】 In this specification, “administering at a lower dose” refers to exposing neurons that overexpress the transgene after gene transfer to a lower dose of the drug than the amount typically administered to a patient to achieve therapeutic efficacy. For example, for the treatment of epilepsy, retigabine has typically been administered at a dose of 600–1200 mg / day; this corresponds to an average plasma concentration of 0.83 μM. We have shown that concentrations below 0.5 μM were effective in vitro in human neurons when using this method. While not limited by theory, we believe this makes it possible to achieve an effective dose of less than 0.5 μM in in vivo administration. 【0077】 In one embodiment, the therapeutically effective dose of the neuromodulatory agent is reduced by up to 100%. For example, by approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 90%, 95%, 90%, or 95%. 【0078】 While not limited by theory, the inventors believe that Kv7.3 overexpression can exert a baseline effect of reducing neuronal excitability even in the absence of neuromodulatory agents. Thus, in some embodiments, the administration of neuromodulatory agents becomes unnecessary. 【0079】 The reduction in the effective therapeutic dose described above is expected to be sufficient to reduce the side effects of neuromodulatory agents to a clinically acceptable level. A clinically acceptable level will be understood by those skilled in the art. 【0080】 In this specification, "enhancing the efficacy of a drug" refers to increasing the therapeutic efficacy of a given drug dose in transdextrins that overexpress the transgene compared to non-transdextrins. 【0081】 As used herein, “reduce adverse effects” means to reduce or eliminate undesirable adverse effects associated with administering a drug to a subject at a particular dose. 【0082】 For example, the neuromodulatory drug retigabine (ezogabine), marketed as an anticonvulsant, has been shown to cause drowsiness, dizziness, tinnitus and vertigo, confusion, and slurred speech at therapeutic doses (600-1200 mg). Less common side effects include tremor, memory loss, difficulty walking, and diplopia. The FDA publicly warned in 2013 that ezogabine may cause ocular abnormalities characterized by blue discoloration of the skin and retinal pigmentation changes. While not limited by theory, the inventors believe that increasing Kv7.3 expression in hyperexcitable neurons associated with epileptic seizures could shift the therapeutic dose range of retigabine to include lower doses, thereby reducing or eliminating the above side effects. In other words, the therapeutic index of retigabine would increase. In fact, studies involving low doses (300-1200 mg) of retigabine have reported fewer dermatological, ophthalmic, and neurological adverse effects (Lerche et al., 2015; Brickel et al., 2020). 【0083】 The “vectors for gene therapy” as used herein are vectors suitable for gene therapy. While AAV capsids have been described herein, it will be understood that other vectors are also suitable and can be used without departing from the scope of the present invention. 【0084】 Typically, this gene therapy is used to treat neurological disorders or conditions. One example of a neurological disorder or condition that can be treated with this gene therapy is spasticity. Spasticity is a neurological symptom that appears in people with various neurological disorders, including, but not limited to, multiple sclerosis, stroke, traumatic brain injury, spinal cord injury, motor neuron disease, and cerebral palsy. Spasticity is caused by motor neurons causing excessive excitation of muscles, which occurs because the motor neurons become "hyperexcited" due to the disease. 【0085】 Neuronal excitability is determined by the frequency of action potentials generated in response to a given stimulus. Increased excitability is indicated by one or more of the scales described below. 【0086】 In this specification, “increased excitability” is defined as a neuron exhibiting at least one of the following characteristics: (a) Increase in the resting voltage (resting membrane potential - RMP) of the neuron (depolarization); this brings it closer to the threshold for generating an action potential; (b) A decrease in the stimulus intensity required to generate an action potential (basic current) in a neuron; (c) An increase in the number of action potentials generated by a given stimulus (frequency) compared to neurons with "normal" levels of excitability; (d) Action potentials continue to be generated even after the stimulus that triggers the firing is discontinued; (e) Increased action potential frequency in neuronal populations (2-100,000) recorded using a microelectrode array (microelectrode array-MEA); (f) An increase in the number or duration of action potential bursts recorded by MEA (an action potential burst is defined as at least five consecutive action potentials with intervals of 100 ms or less between each action potential). 【0087】 For example, the RMP of an overexcitable neuron may be -55mV, which is roughly 15mV higher in the positive direction than what is expected for the neuron and is closer to the threshold for action potential generation (approximately -40mV). If a stimulus intensity of 50 picoamperes (pA) generates an action potential (base current = 50pA) in a normal neuron, then 25pA is sufficient for an overexcitable neuron (base current = 25pA). Similarly, if a stimulus intensity of 100pA causes a normal neuron to generate 10 action potentials in 1 second (frequency = 10Hz), then an overexcitable neuron will generate 20 action potentials at the same stimulus intensity (frequency = 20Hz). 【0088】 Furthermore, hyperexcitable neurons often continue firing action potentials even after the cessation of stimulation, whereas this persistence does not occur in normal neurons. Such cases are observed in spasticity, which causes neurons to fire more action potentials for longer periods; and they also tend to fire in response to weaker stimuli. 【0089】 Typically, the gene therapy vector is an AAV vector containing a nucleotide sequence encoding the Kv7.3 ion channel subunit. The ion channel subunit may be the wild type encoded by SEQ ID NO: 1, or a variant encoded by SEQ ID NO: 2, or an equivalent sequence encoded by an equivalent sequence encoding the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4, respectively. 【0090】 The AAV virus particles described herein are adeno-associated virus particles containing a single-stranded DNA genome incorporated within a viral envelope, and are transdependent to cells (e.g., target neurons). In particular, examples of AAV virus particles described herein include nucleotide sequences according to SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the virus particles may lack replication-encoding nucleotides; that is, the virus particles are replication-deficient. 【0091】 AAV is a small virus belonging to the genus Depend Parvovirus, containing single-stranded DNA up to approximately 4.9 Kb in length. The AAV genome contains three capsid proteins, VP1, VP2, and VP3, all translated from a single mRNA via alternative splicing. In the wild type, multiple serotypes have been identified for AAV; wild-type serotypes tend to infect multiple tissues and cell types, but each serotype has a unique capsid gene sequence and therefore differs in its targeting. These serotypes are denoted by numbers, such as AAV1, AAV2, etc. It has been shown that modification of the capsid sequence using DNA recombination can create non-natural sequences that have properties to fit specific cells or tissues and possess targeting (or antitargeting) to the same, and that evade the immune system (Vandenberghe et al., 2009). The “AAV virus particles” used herein selectively transduce target neurons (such as motor neurons). 【0092】 As used herein, “AAV capsid” refers to the capsid of adeno-associated virus. It is known that the AAV capsid exhibits remarkable selectivity due to its composition and is capable of post-translational modification (the AAV capsid is composed of a mixture of VP1, VP2, and VP3, with 60 monomers arranged icosahedral symmetrically in a 1:1:10 ratio). The AAV capsid protein contains 12 hypervariable surface regions, but its genome generally contains highly conserved replication and structural genes across serotypes. 【0093】 The "AAV capsid" as defined herein can selectively transduce into specific cell types, such as neurons, rather than muscle tissue, and can also selectively transduce into target neurons rather than other neurons, and even into specific parts of neurons (such as axons). 【0094】 For use, an AAV capsid (viral particle) containing a nucleotide sequence according to one of sequence numbers 1-8 is introduced to the target; the viral particle infects the target neuron that will become a transduction neuron. The transduction neuron overexpresses the amino acid sequence encoded by the nucleotide sequence, which then associate to form a functional Kv7.3-containing ion channel (either homomer or heteromer). Generally, it takes approximately three weeks to deliver the viral particle to a target that will overexpress the functional ion channel. Once a suitable time has elapsed for the transduction neuron to overexpress the ion channel, the target can be treated with a lower dose of a neuromodulatory agent such as retigabine. 【0095】 In this specification, "selectively transducing" refers to the transduction ability of an AAV capsid to selectively transduce one cell type with preferential treatment over other cell types. This specificity may, for example, be preferential to neurons over muscle cells, or it may be preferential to motor neurons over sensory neurons. 【0096】 In this specification, “subject” means human or non-human mammal, which typically has a neurological disorder associated with increased neuronal excitability. 【0097】 In some embodiments, the subject has neurological disorders associated with dysfunction of the Kv7.3 ion channel. 【0098】 In some embodiments, the subject has symptoms characterized by neuronal hyperexcitability, which may or may not be related to dysfunction of Kv7.3 ion channels. 【0099】 In some embodiments, the subject has neurological disorders and / or neurological symptoms characterized by neuronal hyperexcitability, which may or may not be related to Kv channel dysfunction. 【0100】 In this specification, "neuropathies associated with increased neuronal excitability" refers to conditions in which excessive neuronal activity causes symptoms associated with the aforementioned neuropathies. Examples include epilepsy, spasticity, benign familial neonatal seizures (BNFS), nociceptive pain, non-nociceptive pain, Parkinson's disease, and multiple sclerosis. 【0101】 In one embodiment, the neurological disorder is related to the excessive excitability of neurons, such as motor neurons. 【0102】 In one embodiment, the neuronal disorder is associated with increased neuronal excitability. 【0103】 In one embodiment, the neurological disorder is: Epilepsy, spasticity, BNFS, nociceptive pain, non-nociceptive pain, Parkinson's disease, multiple sclerosis, Selected from. 【0104】 As used herein, “to treat” or “to treat” means regression of a condition, improvement or reduction of symptoms associated with a condition, or prevention of further onset / worsening of a condition. 【0105】 The term "treatment" includes combination therapies and combination treatments (e.g., sequential or simultaneous combination therapies) that combine two or more treatments or therapies. For example, the vector may be administered the day before or earlier than the administration of the neuromodulatory agent to the subject. 【0106】 As used herein, “lower dose” refers to a dose lower than the dose considered effective for the subject (or a lower portion of the effective range), and therefore a lower dose that does not cause side effects or reduces the severity of side effects while maintaining therapeutic efficacy. 【0107】 As used herein, “therapeutic dose” means the amount of the compound that is effective in treating the specified disorder or condition. 【0108】 In some embodiments, overexpressed Kv7.3 channels may associate to form heteromers such as Kv7.2 / 7.3 heteromers or Kv7.3 / 7.5 heteromers. Since the ratios of the subunits of these heteromers may differ, the method described herein may be fine-tuned. 【0109】 Similarly, in some embodiments, the ratio of the wild-type to the mutant Kv7.3 subunit may differ, so the method may be fine-tuned. This can apply to both heteromers and homomers. 【0110】 In some embodiments, overexpressed Kv7.3 channels associate to form homomers. 【0111】 Since allogeneic Kv7.3 channels may be slightly conductive, they have minimal effect on the baseline excitability of transduction neurons; however, the neuromodulatory agent can still act on transduction neurons at lower doses. 【0112】 In this specification, "base current" refers to the minimum current injection value required to sufficiently depolarize the neuron membrane potential in order to generate a single action potential. 【0113】 In this specification, “resting membrane potential” refers to the potential difference recorded between the intracellular and extracellular compartments of a neuron in the absence of current injection. This voltage value is recorded using electrophysiological techniques such as whole-cell patch clamp. 【0114】 In this specification, "shifting the resting membrane potential (RMP) of a target neuron from the threshold" means that the voltage value in the neuron's resting state is more negative (hyperpolarized), and therefore a stronger stimulus intensity is required to depolarize the voltage to a level where an action potential is generated. For example, a typical RMP of a neuron may be -65mV, which means that a current (stimulus) of 500pA is required to sufficiently depolarize the neuron to reach the threshold membrane potential (-45mV) that generates an action potential. If the RMP is more negative (-75mV), it is further from the threshold, and therefore more current injection is required to reach the threshold and generate an action potential. 【0115】 In this specification, "reducing the excitability of target neurons" means at least one of the following: (a) Hyperpolarize the resting membrane potential of the neuron, moving it further away from the threshold for generating an action potential; (b) Increase the stimulus intensity required to generate an action potential (basic current) in the neuron; (c) Reduce the number of action potentials (AP frequency) generated by a given electrical or chemical stimulus; (d) Reduce or eliminate the persistence of action potentials after the cessation of the stimulus that triggers the firing; (e) Reduce the frequency of spontaneous action potentials in the absence of stimulation; (f) Reduce the duration or frequency of spontaneous action potential bursts (a burst is defined as at least five consecutive action potentials at intervals of 100 ms or less). 【0116】 Decreased excitability can be measured using whole-cell patch-clamp electrophysiology in current-clamp mode. This measurement is performed by injecting a square current pulse with increasing intensity and recording the voltage change and the frequency of action potential generation during the current pulse duration. The resting membrane potential is measured under stable conditions without current injection (e.g., without voltage changes exceeding 1-2 mV). The baseline current is measured as the minimum input current in a given time to generate at least one action potential. The frequency of action potentials is measured as the number of action potentials generated within a specific time, which is usually equal to the duration of the current pulse injected into the neuron. 【0117】 Decreased excitability can also be measured using a microelectrode array (MEA); this microelectrode array consists of a well plate containing multiple electrode contacts located on the substrate surface of each well. Since the neuronal population is grown while in contact with the electrodes in each well, the activity of the neurons can be recorded as extracellular action potentials (spikes). Neurons grown on an MEA generate spontaneous action potentials, which are often called spikes. A series of five or more spikes recorded at intervals of 100 ms or less is called a burst. The frequency of spontaneous spikes and burst characteristics (burst frequency, duration, etc.) may be used to determine the excitability of neurons. 【0118】 In some embodiments, upregulation of the Kv7.3 ion channel subunit in transduction neurons reduces the activity of cells innervated by the axon terminal of the same transduction neuron. For example, reduced gastrocnemius muscle activity, as measured by electromyography (EMG), results from upregulation of the Kv7.3 ion channel subunit in motor neurons of the gastrocnemius muscle. The reduction in muscle activity has one or more of the following characteristics: A decrease in the intensity of compound muscle movements induced by stimulation of peripheral afferent nerve fibers; a reduction in the duration of sustained EMG activity after afferent nerve fiber stimulation; a decrease in the frequency of spontaneous EMG activity; a reduction in the duration of spontaneous EMG activity; a decrease in the intensity of spontaneous EMG activity; a reduction in muscle rigidity as assessed by a skilled clinician; and an increase in the range of motion at the joint to which the target muscle attaches, as assessed by a skilled clinician. 【0119】 The present invention also includes a therapy comprising injecting an AAV viral vector comprising the nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:2 into the target brain or spinal cord, or a therapy by intramuscular injection; these AAV viral vectors enable transduction into target neurons. 【0120】 The expression of the nucleotide sequence leads to the overexpression of the Kv7.3 ion channel in these neurons. The subject can then be treated with a lower dose of the neuromodulator than previously required to achieve a therapeutic effect. This lower dose reduces toxicity by decreasing the occurrence of side effects. The therapeutic objectives are achieved, namely, the alleviation of symptoms in patients suffering from diseases caused by hyperexcitable neurons, and / or the improvement of the patient's quality of life, without causing (or reducing the severity of) undesirable side effects. 【0121】 The methods disclosed herein allow for the effective use of drugs with extremely strong side effects simply by enabling lower doses that are effective for specific cells. 【0122】 In some embodiments, the method includes a gene therapy vector or AAV virus particle that retrogradely infects neurons for introducing genetic material into neurons for the purpose of treating diseases caused by or related to Kv7 ion channel dysfunction. 【0123】 In the case of A315T, a channel with high membrane insertion rate and conductivity, it is likely that only a low expression level is necessary for efficacy. This means that using this virus allows for delivery at lower doses while reducing the probability of immunogenicity and lowering costs. 【0124】 In one embodiment, the method is an ex vivo method. 【0125】 In this specification, please understand that "comprising" means "including". 【0126】 Aspects of the present invention that include specific elements are also intended to be extended to other embodiments that "consist" or "consist essentially" of the relevant elements. 【0127】 Where technically appropriate, embodiments of the present invention may be combined. 【0128】 In this specification, embodiments are described as comprising certain features / elements. The disclosure also extends to other embodiments comprising such features / elements or other embodiments comprising essentially the same. 【0129】 In this specification, "approximately" means within ±10% of the average value. 【0130】 Technical references, such as patents and applications, are incorporated herein by reference. 【0131】 The embodiments specifically and expressly listed herein may form the basis for disclaimers, either individually or in combination with one or more further embodiments. 【0132】 Examples Materials and methods cell culture Human iPSC sensory neuron progenitor cells (Axol Bioscience) were thawed and seeded at low density onto monolayer rat cortical astrocytes on poly-D-lysine coated coverslips. To promote the maturation of the cells into a differentiated neuronal phenotype, the cultures were grown in NbActiv4 (Neurobasal / B 27 supplemented with MAXIMIZER, GDNF, NGF, BDNF, and NT3). 【0133】 AAV On day 7 of in vitro testing, pAAV-CMV-EGFP, pAAV-CMV-hKCNQ3-T2A-EGFP, and pAAV-CMV-hKCNQ3(A315T)-T2A-EGFP were added to wells containing nerve cultures. Patch clamp measurements were performed and recorded either one week later (DIV15-16) or two weeks later (DIV21-23). 【0134】 Patch-clamp electrophysiology The standard patch-clamp method was used. The external measurement solution consisted of 140 mM NaCl, 25 mM KCl, 2 mM CaCl2, 3 mM MgCl2, 10 mM glucose, 10 mM HEPES, and pH 7.3. The internal measurement recording solution consisted of 120 mM potassium gluconate, 20 mM KCl, 3 mM MgCl2, 2.5 mM EGTA, 0.5 mM CaCl, 2.4 mM Na2-ATP, 0.3 mM Li GTP, 10 mM HEPES, and pH 7.3. Once all cells were positioned, the passive membrane properties (capacitance and resistance) of all cells were measured using voltage clamp mode. All excitability measurements were performed using current clamp mode. 【0135】 Measurement recording using a multi-electrode array By growing neurons in a 24-well plate containing 16 electrodes per well using the method described above, the neurons were brought into direct contact with individual electrodes. Then, after 30 days, spontaneous firing activity was recorded. Recordings were made at baseline (without the drug), and then recorded while increasing the concentration of the activating agent. 【0136】 Mouse spasticity model In neonatal mice (day 0 postnativity), the spinal cord was completely transversed between the 9th and 10th thoracic somites to induce spasticity. During this surgery, AAV was injected into both calf muscles. 【0137】 EMG Records 10–14 days after spinal cord injury, EMG electrodes were inserted into the injected muscle, and electrical activity was recorded before and after intraperitoneal administration of the activating agent (retigabine). Different doses were tested on different days. 【0138】 Patch-clamp electrophysiology of mouse spinal cord sections The spinal cord was harvested under euthanasia anesthesia and sectioned into 300 μm sections using a vibratome. Then, patch-clamp measurements of motor neurons expressing GFP were performed and recorded using the method described above (patch-clamp electrophysiology). 【0139】 item [Item 1] A method for enhancing the effectiveness of neuromodulatory drugs; (a) A nucleotide sequence that encodes a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8 via a vector; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the drug into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering the neuromodulatory agent at a lower dose, Methods that include... [Item 2] A method for reducing the side effects of neuromodulatory drugs; (a) A nucleotide sequence that encodes a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8 via a vector; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the drug into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering the neuromodulatory agent at a lower dose, Methods that include... [Item 3] A method to increase the therapeutic index of neuromodulatory drugs; (a) A nucleotide sequence that encodes a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8 via a vector; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the drug into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering the neuromodulatory agent, Methods that include... [Item 4] A method to reduce the excitability of neurons; (a) A nucleotide sequence that encodes a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8 via a vector; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the substance into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering a neuromodulatory agent selectively; Methods that include... [Item 5] A method for treating neurological disorders; (a) A nucleotide sequence encoding a Kv7.3 ion channel subunit, containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A step of administering a therapeutically effective amount of a gene therapy vector containing the specified substance to a target in need; (b) a step of upregulating the expression of the Kv7.3 ion channel subunit in a target neuron to form a functional ion channel; and (c) A process for administering a neuromodulatory agent known to target the Kv7.3 ion channel at a lower dose. Methods that include... [Item 6] A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector containing; When this vector is used to transduce target neurons, the expression of the Kv7.3 ion channel subunit is upregulated, and a functional ion channel is formed. Gene therapy vector. [Item 7] A gene therapy vector that conforms to item 6; Here, the vector selectively achieves transduction into the target neuron. Gene therapy vector. [Item 8] AAV virus particle vector for gene therapy; The vector is: (a) AAV capsids that selectively induce target neurons; and (b) A nucleotide sequence encoding a Kv7.3 ion channel subunit containing any sequence from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, Includes; When this vector is used to transduce target neurons, the expression of the Kv7.3 ion channel subunit is upregulated, and a functional ion channel is formed. Gene therapy vector. [Item 9] AAV capsids that selectively induce plasma delivery to target neurons; and A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector containing; This method is used to upregulate the expression of the Kv7.3 ion channel subunit by the target neuron, thereby enabling the formation of functional ion channels in subjects suffering from neurological disorders associated with increased neuronal excitability. Gene therapy vector. [Item 10] AAV capsids that selectively induce plasma delivery to target neurons; Furthermore A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO:1 to SEQ ID NO:8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector for use in the treatment of subjects with neurological disorders, including; When this vector is used to transduce target neurons, the expression of Kv7.3 ion channels by the target neurons is upregulated, thereby enabling the effective administration of neuromodulatory drugs to the target at lower doses. Gene therapy vector. [Item 11] A method according to any one of items 1-5 or a gene therapy vector according to item 10; Here, the neuromodulatory agent: Retigabin / Ezogabin and its derivatives, BHV-7000 and Xen496, Xen1101, Flupirtin, Diclofenac, BMS-204352, Meclofenamic acid, ETX-123 and Linopyrdin, Selected from, Methods or vectors for gene therapy. [Item 12] A method or gene therapy vector according to any of the above items; Here, the Kv7.3 ion channel is expressed as a Kv7.3 homomer, a Kv7.2 / 7.3 heteromer, or a Kv7.3 / 7.5 heteromer. Methods or vectors for gene therapy. [Item 13] A method or gene therapy vector according to item 12; Here, the Kv7.3 ion channel is functionally expressed as a homomer. Methods or vectors for gene therapy. [Item 14] A method or gene therapy vector according to any of the above items; Here, the target neuron is a motor neuron. Methods or vectors for gene therapy. [Item 15] A method or vector for gene therapy according to item 5 or any one of items 9-14; Here, the neurological disorder is related to increased neuronal excitability. Methods or vectors for gene therapy. [Item 16] A method or vector for gene therapy according to item 5 or any one of items 9-15; Here, the neurological disorder is selected from the group consisting of epilepsy, spasticity, BNFS, Parkinson's disease, nociceptive pain, and non-nociceptive pain. Methods or vectors for gene therapy. [Item 17] A method or gene therapy vector according to any of the above items; Here, by upregulating the Kv7.3 ion channel subunit, the baseline current of the target neuron is reduced in the presence of the neuromodulatory agent. Methods or vectors for gene therapy. [Item 18] A method or gene therapy vector according to any of the above items; Here, by upregulating the Kv7.3 ion channel subunit, the resting membrane potential of the target neuron is shifted away from the threshold in the presence of the neuromodulatory agent. Methods or vectors for gene therapy. [Item 19] A method or gene therapy vector according to any of the above items; Here, by upregulating the Kv7.3 ion channel subunit, the excitability of the target neuron is reduced. Methods or vectors for gene therapy. [Item 20] A method or gene therapy vector according to any of the above items; By upcontrolling the Kv7.3 ion channel subunit, the excitability of multiple target neurons is reduced when recording using a microelectrode array (MEA) device. Methods or vectors for gene therapy. [Item 21] A method or gene therapy vector according to any of the above items; Here, by upregulating the Kv7.3 ion channel subunit by the target neuron, the activity of cells innervated by the axon terminal of the target neuron is reduced. Methods or vectors for gene therapy. [Item 22] Methods 19-21; In the presence of the neuromodulatory agent, a decrease in the excitability of the target neuron, or a decrease or enhancement in the activity of cells innervated by the axon terminal of the target neuron, occurs. method. [Item 23] Uses of gene therapy vectors in accordance with items 6-10; In the preparation of therapeutic agents used to treat conditions that are improved by upregulating the expression of the Kv7.3 ion channel, Purpose. [Item 24] Uses of gene therapy vectors in accordance with items 6-10 in the preparation of therapeutic agents for the treatment of neurological disorders associated with increased neuronal excitability; Herein the nerve damage occurred: Epilepsy, spasticity, BNFS, Parkinson's disease, ALS, nociceptive pain and non-nociceptive pain, Selected from the group consisting of, Purpose. [Item 25] Uses of gene therapy vectors in accordance with items 6-10; In the preparation of therapeutic drugs used as antispasmodics, Purpose. References Gomez-Posada, JC et al. A Pore Residue of the KCNQ3 Potassium M-Channel Subunit Controls Surface Expression. Journal of Neuroscience 30, 9316-9323 (2010). Zaika, O., Hernandez, C. C., Bal, M., Tolstykh, G. P. & Shapiro, M. S. Determinants within the Turret and Pore-Loop Domains of KCNQ3 K+ Channels Governing Functional Activity. Biophys J 95, 5121-5137 (2008). Musella, S. et al. Beyond Retigabine: Design, Synthesis, and Pharmacological Characterization of a Potent and Chemically Stable Neuronal Kv7 Channel Activator with Anticonvulsant Activity. J Med Chem 65, 11340-11364 (2022). Corbin-Leftwich, A. et al. Retigabine holds KV7 channels open and stabilizes the resting potential. Journal of General Physiology 147, 229-241 (2016). Gunthorpe, M., Large, C., Epilepsia, R. S.- & 2012, undefined. The mechanism of action of retigabine (ezogabine), a first‐in‐class K+ channel opener for the treatment of epilepsy. Wiley Online Library 53, 412-424 (2012). French, J. A. et al. Randomized, double-blind, placebo-controlled trial of ezogabine (retigabine) in partial epilepsy. Neurology 76, 1555-1563 (2011). Villalba-Galea, C. A. Modulation of KV7 Channel Deactivation by PI(4,5)P2. Front Pharmacol 11, (2020). Brickel, N. et al. Safety of retigabine in adults with partial-onset seizures after long-term exposure: focus on unexpected ophthalmological and dermatological events. Epilepsy & Behavior 102, 106580 (2020). Lerche, H. et al. Efficacy and safety of ezogabine / retigabine as adjunctive therapy to specified single antiepileptic medications in an open-label study of adults with partial-onset seizures. Seizure 30, 93-100 (2015). Harris, J.A. and Murphy, J.A. Retigabine (ezogabine) as add-on therapy for partial-onset seizures: an update for clinicians. Ther Adv Chronic Dis. 2011 Nov; 2(6): 371-6. Array Accession number: 1 - wild-type Kv7.3 >ENA|AAC96101|AAC96101.1 ホモサピエンス(ヒト)カリウムチャネル ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGGCCACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Accession Number: 2-Kv7.3 A315T >ENA|AAC96101|AAC96101.1 Homo sapiens (Human) potassium channel A315T ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGACCACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Accession number: 3 - Kv7.3 A315S >ENA|AAC96101|AAC96101.1 Homo sapiens (human) potassium channel A315S ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGTCCACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Accession Number: 4-Kv7.3 A315V >ENA|AAC96101|AAC96101.1 Homo sapiens (Human) potassium channel A315V ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGGTCACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Accession number: 5-Kv7.3 A315C >ENA|AAC96101|AAC96101.1 Homo sapiens (human) potassium channel A315C ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGTGCACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Accession No.: 6-Kv7.3 A315N >ENA|AAC96101|AAC96101.1 Homo sapiens (human) potassium channel A315N ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGAACACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Accession number: 7-Kv7.3 A315Q >ENA|AAC96101|AAC96101.1 Homo sapiens (human) potassium channel A315Q ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGCAGACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Accession number: 8-Kv7.3 A315Y >ENA|AAC96101|AAC96101.1 Homo sapiens (Human) potassium channel A315Y ATGGGGCTCAAGGCGCGCAGGGCGGCGGGGGCGGCTGGCGGCGGCGGCGACGGGGGCGGC GGAGGCGGCGGGGCGGCTAACCCAGCCGGAGGGGACGCGGCGGCGGCCGGCGACGAGGAG CGGAAAGTGGGGCTGGCGCCCGGCGACGTGGAGCAAGTCACCTTGGCGCTCGGGGCCGGA GCCGACAAAGACGGGACCCTGCTGCTGGAGGGCGGCGGCCGCGACGAGGGGCAGCGGAGG ACCCCGCAGGGCATCGGGCTCCTGGCCAAGACCCCGCTGAGCCGCCCAGTCAAGAGAAAC AACGCCAAGTACCGGCGCATCCAAACTTTGATCTACGACGCCCTGGAGAGACCGCGGGGC TGGGCGCTGCTTTACCACGCGTTGGTGTTCCTGATTGTCCTGGGGTGCTTGATTCTGGCT GTCCTGACCACATTCAAGGAGTATGAGACTGTCTCGGGAGACTGGCTTCTGTTACTGGAG ACATTTGCTATTTTCATCTTTGGAGCCGAGTTTGCTTTGAGGATCTGGGCTGCTGGATGT TGCTGCCGATACAAAGGCTGGCGGGGCCGACTGAAGTTTGCCAGGAAGCCCCTGTGCATG TTGGACATCTTTGTGCTGATTGCCTCTGTGCCAGTGGTTGCTGTGGGAAACCAAGGCAAT GTTCTGGCCACCTCCCTGCGAAGCCTGCGCTTCCTGCAGATCCTGCGCATGCTGCGGATG GACCGGAGAGGTGGCACCTGGAAGCTTCTGGGCTCAGCCATCTGTGCCCACAGCAAAGAA CTCATCACGGCCTGGTACATCGGTTTCCTGACACTCATCCTTTCTTCATTTCTTGTCTAC CTGGTTGAGAAAGACGTCCCAGAGGTGGATGCACAAGGAGAGGAGATGAAAGAGGAGTTT GAGACCTATGCAGATGCCCTGTGGTGGGGCCTGATCACACTGTACACCATTGGCTATGGA GACAAGACACCCAAAACGTGGGAAGGCCGTCTGATTGCCGCCACCTTTTCCTTAATTGGC GTCTCCTTTTTTGCCCTTCCAGCGGGCATCCTGGGGTCCGGGCTGGCCCTCAAGGTGCAG GAGCAACACCGTCAGAAGCACTTTGAGAAAAGGAGGAAGCCAGCTGCTGAGCTCATTCAG GCTGCCTGGAGGTATTATGCTACCAACCCCAACAGGATTGACCTGGTGGCGACATGGAGA TTTTATGAATCAGTCGTCTCTTTTCCTTTCTTCAGGAAAGAACAGCTGGAGGCAGCATCC AGCCAAAAGCTGGGTCTCTTGGATCGGGTTCGCCTTTCTAATCCTCGTGGTAGCAATACT AAAGGAAAGCTATTTACCCCTCTGAATGTAGATGCCATAGAAGAAAGTCCTTCTAAAGAA CCAAAGCCTGTTGGCTTAAACAATAAAGAGCGTTTCCGCACGGCCTTCCGCATGAAAGCC TACGCTTTCTGGCAGAGTTCTGAAGATGCCGGGACAGGTGACCCCATGGCGGAAGACAGG GGCTATGGGAATGACTTCCCCATCGAAGACATGATCCCCACCCTGAAGGCCGCCATCCGA GCCGTCAGAATTCTACAATTCCGTCTCTATAAAAAAAAATTCAAGGAGACTTTGAGGCCT TACGATGTGAAGGATGTGATTGAGCAGTATTCTGCCGGGCATCTCGACATGCTTTCCAGG ATAAAGTACCTTCAGACGAGAATAGATATGATTTTCACCCCTGGACCTCCCTCCACGCCA AAACACAAGAAGTCTCAGAAAGGGTCAGCATTCACCTTCCCATCCCAGCAATCTCCCAGG AATGAACCATATGTAGCCAGACCATCCACATCAGAAATCGAAGACCAAAGCATGATGGGG AAGTTTGTAAAAGTTGAAAGACAGGTTCAGGACATGGGGAAGAAGCTGGACTTCCTCGTG GATATGCACATGCAACACATGGAACGGTTGCAGGTGCAGGTCACGGAGTATTACCCAACC AAGGGCACCTCCTCGCCAGCTGAAGCAGAGAAGAAGGAGGACAACAGGTATTCCGATTTG AAAACCATCATCTGCAACTATTCTGAGACAGGCCCCCCGGAACCACCCTACAGCTTCCAC CAGGTGACCATTGACAAAGTCAGCCCCTATGGGTTTTTTGCACATGACCCTGTGAACCTG CCCCGAGGGGGACCCAGTTCTGGAAAGGTTCAGGCAACTCCTCCTTCCTCAGCAACAACG TATGTGGAGAGGCCCACGGTCCTGCCTATCTTGACTCTTCTCGACTCCCGAGTGAGCTGC CACTCCCAGGCTGACCTGCAGGGCCCCTACTCGGACCGAATCTCCCCCCGGCAGAGACGT AGCATCACGCGAGACAGTGACACACCTCTGTCCCTGATGTCGGTCAACCACGAGGAGCTG GAGAGGTCTCCAAGTGGCTTCAGCATCTCCCAGGACAGAGATGATTATGTGTTCGGCCCC AATGGGGGGTCGAGCTGGATGAGGGAGAAGCGGTACCTCGCCGAGGGTGAGACGGACACA GACACGGACCCCTTCACGCCCAGCGGCTCCATGCCTCTGTCGTCCACAGGGGATGGGATT TCTGATTCAGTATGGACCCCTTCCAATAAGCCCATTTAA Sequence ID: 9 - Wild Type Kv7.3 >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRRSLRFLQILRMMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLATIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI Sequence ID: 10-Kv7.3 A315T >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 A315T MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRSLRFLQILRMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLTTIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI Sequence ID: 11-Kv7.3 A315S >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 A315S MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRRSLRFLQILRMMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLSTIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI Sequence ID: 12-Kv7.3 A315V >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 A315V MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRRSLRFLQILRMMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLVTIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI Sequence ID: 13-Kv7.3 A315C >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 A315C MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRRSLRFLQILRMMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLCTIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI Sequence ID: 14-Kv7.3 A315N >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 A315N MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRRSLRFLQILRMMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLNTIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI Sequence ID: 15-Kv7.3 A315Q >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 A315Q MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRRSLRFLQILRMMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLQTIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI Sequence ID: 16-Kv7.3 A315Y >sp|O43525|KCNQ3_HUMAN Voltage-gated potassium channel subfamily KQT member 3 OS=Homo sapiens OX=9606 GN=KCNQ3 PE=1 SV=2 A315Y MGLKARRAAGAAGGGGDGGGGGGGAANPAGGDAAAAGDEERKVGLAPGDVEQVTLALGAG ADKDGTLLLEGGGRDEGQRRTPQGIGLLAKTPLSRPVKRNNAKYRRIQTLIYDALERPRG WALLYHALVFLIVLGCLILAVLTTFKEYETVSGDWLLLLETFAIFIFGAEFALRIWAAGC CCRYKGWRGRLKFARKPLCMLDIFVLIASVPVVAVGNQGNVLATSLRRSLRFLQILRMMLRM DRRGGTWKLLGSAICAHSKELITAWYIGFLTLILSSFLVYLVEKDVPEVDAQGEEMKEEF ETYADALWWGLITLYTIGYGDKTPKTWEGRLIAATFSLIGVSFFALPAGILGSGLALKVQ EQHRQKHFEKRRKPAAELIQAAWRYYATNPNRIDLVATWRFYESVVSFPFFRKEQLEAAS SQKLGLLDRVRLSNPRGSNTKGKLFTPLNVDAIEESPSKEPKPVGLNNKERFRTAFRMKA YAFWQSSEDAGTGDPMAEDRGYGNDFPIEDMIPTLKAAIRAVRILQFRLYKKKFKETLRP YDVKDVIEQYSAGHLDMLSRIKYLQTRIDMIFTPGPPSTPKHKKSQKGSAFTFPSQQSPR NEPYVARPSTSEIEDQSMMGKFVKVERQVQDMGKKLDFLVDMHMQHMERLQVQVTEYYPT KGTSSPAEAEKKEDNRYSDLKTIICNYSETGPPEPPYSFHQVTIDKVSPYGFFAHDPVNL PRGGPSSGKVQATPPSSATTYVERPTVLPILTLLDSRVSCHSQADLQGPYSDRISPRQRR SITRDSDTPLSLMSVNHEELERSPSGFSISQDRDDYVFGPNGGSSWMREKRYLAEGETDT DTDPFTPSGSMPLSSTGDGISDSVWTPSNKPI

Claims

[Claim 1] A method for enhancing the effectiveness of neuromodulatory drugs; (a) via vector, A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the drug into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering the neuromodulatory agent at a lower dose, including, method. [Claim 2] A method for reducing the side effects of neuromodulatory drugs; (a) via vector, A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the drug into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering the neuromodulatory agent at a lower dose, including, method. [Claim 3] A method to increase the therapeutic index of neuromodulatory drugs; (a) via vector, A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the drug into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering the neuromodulatory agent, including, method. [Claim 4] A method to reduce the excitability of neurons; (a) via vector, A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, The process of introducing the drug into the target neuron; (b) A step of overexpressing the ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering a neuromodulatory agent selectively, including, method. [Claim 5] A method of treating neurological disorders; (a) A nucleotide sequence encoding a Kv7.3 ion channel subunit, which includes one of the sequences of SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A step of administering a therapeutically effective amount of a gene therapy vector containing the gene to a target in need; (b) A step of upregulating the expression of the Kv7.3 ion channel subunit in the target neuron to form a functional ion channel; and (c) A step of administering a neuromodulatory agent known to target the Kv7.3 ion channel at a lower dose. including, method. [Claim 6] A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector containing; When this vector is used to transduce target neurons, the expression of the Kv7.3 ion channel subunit is upregulated, and a functional ion channel is formed. Gene therapy vector. [Claim 7] A gene therapy vector according to claim 6; Here, the vector selectively achieves transduction into the target neuron. Gene therapy vector. [Claim 8] AAV virus particle vector for gene therapy; The vector is: (a) AAV capsids that selectively induce target neurons; and (b) A nucleotide sequence encoding a Kv7.3 ion channel subunit, which includes one of the sequences of SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, Including; When this vector is used to transduce target neurons, the expression of the Kv7.3 ion channel subunit is upregulated, and a functional ion channel is formed. AAV virus particle vector for gene therapy. [Claim 9] AAV capsids that selectively induce plasma delivery to target neurons; and A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector containing; This method is used to upregulate the expression of the Kv7.3 ion channel subunit by the target neuron, thereby enabling the formation of a functional ion channel in a subject suffering from a neurological disorder associated with increased neuronal excitability. Gene therapy vector. [Claim 10] AAV capsids that selectively induce plasma delivery to target neurons; and A nucleotide sequence encoding a Kv7.3 ion channel subunit containing one of the sequences from SEQ ID NO: 1 to SEQ ID NO: 8; or A nucleotide sequence encoding the amino acid sequence of any of the Kv7.3 ion channel subunits from SEQ ID NO: 9 to SEQ ID NO: 16, A gene therapy vector for use in the treatment of subjects with neurological disorders, including; When this vector is used to transduce target neurons, the expression of Kv7.3 ion channels by the target neurons is upregulated, thereby enabling the effective administration of neuromodulatory drugs to the target at lower doses. Gene therapy vector. [Claim 11] A method according to any one of claims 1 to 5 or a gene therapy vector according to claim 10; Here, the neuromodulatory agent: Retigavin / ezogavin and its derivatives, BHV-7000 and Xen496, Xen1101, flupirtin, diclofenac, BMS-204352, meclofenamic acid, ETX-123 and linopyrdin, Selected from, Methods or vectors for gene therapy. [Claim 12] A method or vector for gene therapy according to any of the above claims; Here, the Kv7.3 ion channel is expressed as a Kv7.3 homomer, a Kv7.2 / 7.3 heteromer, or a Kv7.3 / 7.5 heteromer. Methods or vectors for gene therapy. [Claim 13] A method or gene therapy vector according to claim 12; Here, the Kv7.3 ion channel is functionally expressed as a homomer. Methods or vectors for gene therapy. [Claim 14] A method or vector for gene therapy according to any of the above claims; Here, the target neuron is a motor neuron. Methods or vectors for gene therapy. [Claim 15] A method or vector for gene therapy according to any one of claims 5 or 9 to 14; Here, the neurological disorder is related to increased neuronal excitability. Methods or vectors for gene therapy. [Claim 16] A method or vector for gene therapy according to any one of claims 5 or 9 to 15; Here, the neurological disorder is selected from the group consisting of epilepsy, spasticity, BNFS, Parkinson's disease, nociceptive pain, and nonnociceptive pain. Methods or vectors for gene therapy. [Claim 17] A method or vector for gene therapy according to any of the above claims; Here, by upregulating the Kv7.3 ion channel subunit, the basal current of the target neuron is reduced in the presence of the neuromodulatory agent. Methods or vectors for gene therapy. [Claim 18] A method or vector for gene therapy according to any of the above claims; Here, by upregulating the Kv7.3 ion channel subunit, the resting membrane potential of the target neuron is shifted away from the threshold in the presence of the neuromodulatory agent. Methods or vectors for gene therapy. [Claim 19] A method or vector for gene therapy according to any of the above claims; Here, by upregulating the Kv7.3 ion channel subunit, the excitability of the target neuron is reduced. Methods or vectors for gene therapy. [Claim 20] A method or vector for gene therapy according to any of the above claims; By controlling the Kv7.3 ion channel subunit upwards, the excitability of multiple target neurons is reduced when recording using a microelectrode array (MEA) device. Methods or vectors for gene therapy. [Claim 21] A method or vector for gene therapy according to any of the above claims; Here, by upregulating the Kv7.3 ion channel subunit by the target neuron, the activity of cells innervated by the axon terminal of the target neuron is reduced. Methods or vectors for gene therapy. [Claim 22] The method according to claims 19 to 21; In the presence of the neuromodulatory agent, a decrease in the excitability of the target neuron, or a decrease or enhancement in the activity of cells innervated by the axon terminal of the target neuron, occurs. method. [Claim 23] Uses of gene therapy vectors according to claims 6 to 10; In the preparation of therapeutic agents used to treat conditions that are improved by upregulating the expression of Kv7.3 ion channels, Use. [Claim 24] An application of a gene therapy vector according to claims 6 to 10 in the preparation of a therapeutic agent for the treatment of neurological disorders associated with increased neuronal excitability; Herein the nerve damage occurred: Epilepsy, spasticity, BNF-FS, Parkinson's disease, ALS, nociceptive pain and non-nociceptive pain, Selected from the group consisting of, Use. [Claim 25] Uses of gene therapy vectors according to claims 6 to 10; In the preparation of therapeutic drugs used as antispasmodics, Use.

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