Biology subjects

Seki, K.

Publications and source records attributed to Seki, K..

At least 19 recordsLinked to original sources

bioRxiv2026

Neural populations in the dorsal and ventral cervical spinal cord share temporal dynamics during reach

Neural population dynamics provide a popular framework for studying motor cortex function. However, whether such dynamics reflect stage-specific computations or are shared across the input and output stages of a sensorimotor circuit is unknown. In the spinal cord, afferent inputs and motor outputs are segregated by anatomical depth. Here we developed depth-resolved Neuropixels recordings from the rat cervical spinal cord during targeted reaching behavior. Across our recordings, population activity was low-dimensional and organized into transient, rotational and target-specific dynamical motifs. Despite differences in single-neuron activity by depth, dorsal and ventral populations expressed nearly identical dynamics across targets and animals. The dominant target-invariant component was strikingly synchronized across depths, whereas other components, including the target-specific component, showed condition-dependent ventral-to-dorsal delays with timing consistent with movement-related feedback. Thus, cervical spinal population dynamics span sensory and motor territories, and may reflect shared coordination of activity rather than purely state-specific local processing.

neuroscience↗

bioRxiv2026

The automated eukaryotic pangenome pipeline EukPan reveals accessory genome differentiation beyond core-gene phylogeny in Aspergillus oryzae

Pangenome analysis reveals recurrent gene-content variation beyond a single reference genome, but its application to eukaryotes is constrained by inconsistent gene annotation. ANNEVO predicts gene models from genome FASTA assemblies without RNA-seq data. We developed EukPan, an automated post-annotation pipeline that standardizes GFF/GTF files, selects representative isoforms, constructs proteomes, infers orthogroups, builds a concatenated single-copy core-protein alignment, and summarizes shared accessory orthogroups while excluding orthogroups detected in only one genome. Applied with ANNEVO to 123 Aspergillus oryzae genomes, EukPan identified 11,245 core and 4,407 shared accessory orthogroups. The core-protein phylogeny broadly recovered the reported A-H classification, whereas accessory-genome analyses clearly separated the 33 group-A strains from the other 90 strains. Directional analysis identified 62 group-A-associated and 158 group-A-depleted orthogroups, with major facilitator superfamily (MFS) transporter and fungal Zn2Cys6 transcription-factor domains prominent in the depleted set. Among 93 orthogroups present in all non-A strains and absent from all group-A strains, 59 mapped to 10 segments of RIB40, the standard A. oryzae reference genome and a non-A (group-F) strain. EukPan therefore enables reproducible, coordinated core- and accessory-pangenome analysis from eukaryotic genome assemblies.

genomics↗

bioRxiv2026

Hierarchical neuronal processing in primary somatosensory cortex during action observation

Voluntary movement requires the central nervous system to transform and integrate visual and somatosensory information into coordinated motor outputs. Although mirror-like neuronal activity during both action execution and observation has been extensively described in premotor, motor, and parietal cortices, it remains unknown whether the primary somatosensory cortex (S1) also participates in the action observation network. Here, we recorded single-unit activity from cytoarchitectonically defined areas 3a, 3b, 1, and 2 in macaque S1 while monkeys either executed or observed grasping movements. Approximately one-third of neurons across S1 modulated their firing during action observation, with the proportion of responsive neurons increasing from area 3 to areas 1 and 2, consistent with the hierarchical organization of somatosensory processing. Most action observation neurons showed congruent activity during action execution and observation, suggesting that these responses may reflect top-down motor-related or integrated visuomotor signals and are unlikely to be explained by visual input alone. The higher prevalence of action observation neurons in areas 1 and 2 suggests that action observation-related signals preferentially influence later stages of somatosensory processing, potentially via cortico-cortical interactions with motor and parietal regions.

neuroscience↗

bioRxiv2026

Fluid amyloid-β (Aβ) biomarkers reflect early β-sheet-rich Aβ deposition during the preclinical stage in Alzheimer's disease model 5XFAD mice

Early detection of disease progression using clinically-relevant biomarkers in animal models is important for mechanistic studies and for developing therapeutics in neurodegenerative diseases including Alzheimers disease (AD). The preclinical stage of AD, when amyloid-{beta} (A{beta}) starts to accumulate before cognitive decline, provides a critical window for disease modification. In humans, decreases in cerebrospinal fluid (CSF) A{beta}42 and the A{beta}42/A{beta}40 ratio in preclinical AD are considered to reflect the preferential sequestration of aggregation-prone A{beta}42 into {beta}-sheet-rich deposition in the brain, with corresponding changes being detectable in plasma. However, the extent to which these biomarker-pathology relationships are recapitulated in AD model mice remains incompletely defined. Here we show that CSF and plasma A{beta}42 and the A{beta}42/A{beta}40 ratio decline with age in parallel with the progression of {beta}-sheet-rich A{beta} deposition in preclinical 5XFAD mice, one of the most widely used AD mouse models, as assessed through monthly profiling of these biomarkers. Notably, the CSF A{beta}42/A{beta}40 ratio showed a negative correlation with {beta}-sheet-rich A{beta} deposition in the brain, whereas CSF A{beta}40 did not show a comparable association. In addition, the plasma A{beta}42/A{beta}40 ratio showed a positive correlation with the CSF A{beta}42/A{beta}40 ratio, suggesting that the plasma A{beta}42/A{beta}40 ratio may also reflect brain A{beta} deposition in this model. The strength of these correlations differed by sex, suggesting that sex-dependent differences in the A{beta} kinetics in this model may influence how closely fluid biomarkers reflect pathological progression. These findings support the potential utility of fluid A{beta} as a pathology-linked, translatable biomarker in preclinical 5XFAD mice. Highlights- Fluid A{beta} biomarkers are associated with early A{beta} deposition in preclinical 5XFAD mice. - The CSF A{beta}42/A{beta}40 ratio negatively correlates with {beta}-sheet-rich brain A{beta} deposition. - The plasma A{beta}42/A{beta}40 ratio positively correlates with the CSF A{beta}42/A{beta}40 ratio. - Monthly profiling defines fluid A{beta} biomarker dynamics in preclinical 5XFAD mice. - Sex differences may affect biomarker-pathology relationships in these mice.

neuroscience↗

bioRxiv2025

Protocol for intra-nerve AAV injection and dorsal root potential recording for optogenetic modulation of the peripheral sensory nerve activity

Optogenetic modulation of peripheral sensory nerve activity holds great potential for the treatment of sensory disorders. Here, we present a protocol for applying optogenetic techniques to peripheral sensory nerves using an adeno-associated virus (AAV) vector. We describe the procedure for gene transduction into dorsal root ganglion neurons via retrograde transport following intra-nerve AAV injection. We then outline a terminal, acute electrophysiological experiment to evaluate optogenetic effects at the level of the dorsal root. For complete details on the use and execution of this protocol, please refer to Kosugi et al1. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=195 SRC="FIGDIR/small/693516v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@5a2de4org.highwire.dtl.DTLVardef@7ad4f0org.highwire.dtl.DTLVardef@32113aorg.highwire.dtl.DTLVardef@10918c9_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

bioRxiv2025

Spinal interneuronal populations encode static hindlimb posture in the cat

Proprioceptive signals from primary afferents reflect changes in single-joint angles, whereas neuronal population in the cerebral cortex represent whole-limb postures. Where and how this transformation emerges along the somatosensory axis from peripheral proprioceptive receptors remains unclear. We simultaneously recorded many lumbosacral spinal neurons in two decerebrate, immobilized cats while a robotic device held the hindlimb at 16 static endpoint positions spanning hip-knee configurations. Using high-density multielectrode recordings, we asked how spinal populations encode static limb state. At the single-neuronal level, activities in a majority of neurons covaried with a single joints angle (hip or knee), a smaller subset showed combined modulation by both joints, and a distinct subset ( single-endpoint neurons) fired selectively at one unique hip-knee configuration near the sampled joint-range limits and was quiescent in adjacent postures. Population analyses revealed a low-dimensional structure: the first two principal components tracked knee and hip angles, respectively, whereas a third component isolated a boundary-aligned, posture-specific pattern, with loadings peaking at the same extreme configurations preferred by single-endpoint neurons. Decoders trained on ensemble activity reconstructed both joint angles and the limbs endpoint position in body-centered coordinates, indicating that the recorded spinal-interneuron populations contain sufficient information to reconstruct whole-limb kinematics. Together, these findings are consistent with a hierarchical organization whereby joint-based representations within spinal-interneuron populations could contribute to the emergence of limb-centered representations in the ascending proprioceptive pathways. The boundary preference of single-endpoint neurons supports a possible categorical coding scheme at workspace limits that may provide spinal "landmarks" for switching control modes, enhancing stability near kinematic extremes, and supporting recalibration of proprioceptive population codes. Key PointsO_LIWe simultaneously recorded many lumbar spinal neurons in two decerebrate, immobilized cats while a robot held the hindlimb at 16 static positions to test how spinal populations encode posture. C_LIO_LIMany neurons varied with a single joint angle (hip or knee), a smaller subset showed combined hip-knee modulation, and a distinct subset was active only at one specific endpoint posture. C_LIO_LIPopulation analyses revealed a low-dimensional structure: the first two principal components tracked knee and hip angles, while a third captured a posture-specific pattern aligned with single-endpoint neurons. C_LIO_LIDecoders trained on ensemble activity reconstructed joint angles and the limbs endpoint, indicating that spinal interneuronal populations contain sufficient information for whole-limb kinematics. C_LIO_LIThese findings are consistent with a hierarchical organization whereby joint-based representations within spinal-interneuron populations could contribute to the emergence of limb-centered representations in the ascending proprioceptive pathways. C_LI

neuroscience↗

bioRxiv2025

Engineered orthogonal translation systems from metagenomic libraries expand the genetic code

Genetic code expansion with non-canonical amino acids (ncAAs) opens new opportunities for the function and design of proteins by broadening their chemical repertoire. Unfortunately, ncAA incorporation is limited both by a small collection of orthogonal aminoacyl-tRNA synthetases (aaRSs) and tRNAs and by low-throughput methods to discover them. Here, we report the discovery, characterization, and engineering of a UGA suppressing orthogonal translation system mined from metagenomic data. We developed an integrated computational and experimental pipeline to profile the orthogonality of >200 tRNAs, test >1,250 combinations of aaRS:tRNA pairs, and identify the AP1 TrpRS:tRNATrpUCA as an orthogonal pair that natively encodes tryptophan at the UGA codon. We demonstrate that the AP1 TrpRS:tRNATrpUCA is highly active in cell-free and cellular contexts. We then use Ochre, a genomically recoded Escherichia coli strain that lacks UAG and UGA codons, to engineer an AP1 TrpRS variant capable of 5-hydroxytryptophan incorporation at an open UGA codon. We anticipate that our strategy of integrating metagenomic bioprospecting with cell-free screening and cell-based engineering will accelerate the discovery and optimization of orthogonal translation systems for genetic code expansion.

synthetic biology↗

bioRxiv2025

Dynamic and context-dependent modulation of proprioceptive input in primate primary motor cortex

During voluntary movement, somatosensory input is attenuated--sensory gating--which may prevent the CNS from being overwhelmed by predictable afferent feedback not essential for motor control. In the cerebral cortex, sensory gating has been demonstrated in primary (S1) and higher-order sensory areas. The primary motor cortex (M1) is the major cortical output relay to the spinal cord and muscles, and proprioceptive feedback is crucial for shaping motor output, much as in S1. However, whether proprioceptive signals to M1 are attenuated during movement, and if so, why, remains unclear. We recorded somatosensory-evoked potentials (SEPs) in M1 of two monkeys performing a wrist task while electrically stimulating muscle and cutaneous afferents innervating forearm extensors and adjacent skin. Both local field potentials and single-neuron recordings revealed significant suppression of muscle-evoked SEPs during Active movement and Static hold phases, providing direct evidence that proprioceptive input to M1 is generally gated during motor execution, as previously reported for cutaneous input. Yet, amidst this suppression, SEPs in a subset of M1 neurons were preserved during Static hold, especially those evoked by antagonist muscle afferents. Because monkeys had to maintain precise joint angles for stable posture, these results suggest that proprioceptive signals from antagonist muscles--likely reflecting spindle activity in lengthening muscles--escape attenuation to provide information essential for joint angle control. Overall, our findings demonstrate that while proprioceptive input to M1 is broadly suppressed during motor output, specific afferent signals from antagonist muscles are selectively maintained in a context-dependent manner to support posture control.

neuroscience↗

bioRxiv2025

A combinatorial mutational map of active non-native protein kinases by deep learning guided sequence design

Mapping protein sequence-function landscapes has either been limited to small steps (only few mutations) or to sequences similar to those already explored by evolution to maintain activity. Here, we overcome both limitations by applying deep-learning guided redesign to a natural protein tyrosine kinase to generate novel, functional sequences with highly combinatorial mutations. Using cell-free assays, we measure the activities and concentrations of 537 redesigned sequences, which differ from the wild-type by an average of 37 mutations while retaining activity in 85% of variants. These sequences sample 436 unique mutations at 76 different positions throughout the kinase domain. A simple regression model identifies key sequence determinants of function and predicts the function of unseen sequences. Our approach demonstrates how integrating deep-learning guided redesign, functional measurement at scale, and interpretable computational modelling enables functional exploration of highly combinatorial and sparse sequence-function landscapes at mutational scales not possible before.

biochemistry↗

bioRxiv2025

Conserved accessory genes link phylogenetically distinct Bacillus subtilis from Japanese and Indian fermented soyfoods

Bacillus subtilis is central to Asian fermented soybean foods, including Japanese natto. To explore the genomic boundaries of B. subtilis var. natto, we conducted a comparative pangenome analysis of 42 strains, including the core natto clade (n=26) and its closest relatives. Our analysis revealed a striking evolutionary paradox centered on a single strain isolated from Indian bekang. Core-genome phylogenetic analysis places this bekang strain clearly outside the tight natto clade, with a Nepalese kinema strainbeing its closest systematic neighbor. In stark contrast, quantitative analysis of accessory gene profiles revealed this single bekang strain is the functional nearest neighbor to the natto clade, sharing a highly conserved accessory gene repertoire. This shared profile defines a "natto-type" adaptive strategy (the "broad-sense natto group," n=27), separating it from other related strains. Analysis of this group-specific repertoire revealed an enrichment of transcriptional regulators and metabolic enzymes. This finding provides a compelling case study (n=1) of polygenic adaptation, suggesting complex evolutionary pathways, such as horizontal gene transfer or selective retention, can drive rapid adaptation across disparate lineages.

genomics↗

bioRxiv2025

Autogenic spinal excitatory circuit ensures skilled hand movements in primates

Skillful hand movements are a hallmark of primates, including humans, requiring sophisticated motor planning and execution. Challenging the longstanding paradigm that emphasizes the dominance of the cerebral cortex1, our study highlights a pivotal role for spinal excitatory reflex circuits in both planning and executing skillful hand movements. Using a combination of experimental approaches with behaving non-human primates and predictive simulation, we identified a group of excitatory spinal interneurons that orchestrate a closed-loop, positive feedback mechanism during voluntary wrist movements. This mechanism is characterized by a bidirectional interaction between inter-neuronal spiking and muscle activity, mediated by motoneuronal efferent signals and proprioceptive afferent signals from the same agonistic muscles. Furthermore, we demonstrate that the temporal profile of muscle activity during movement execution, including amplitude and duration, is pre-determined during motor planning at the spinal interneurons, functioning as a force-feedback gain within the excitatory circuit. These findings suggest that autogenic spinal excitatory circuits play a predominant role in shaping overall muscle activation during motor execution, provided the proper reflex gain is pre-set by higher neural systems during motor planning. Together, our findings provide a cellular-level perspective on how spinal reflex loops contribute to skilled voluntary movements in primates, extending over a century of spinal reflex research2.

neuroscience↗

bioRxiv2025

Establishment of a second-generation transgenic marmoset model of polyglutamine disease recapitulating neurological symptoms and pathology

Neurodegenerative diseases, including polyglutamine diseases, remain a major clinical challenge, partly because of limited animal models that recapitulate human disease. Here, we describe a second-generation transgenic marmoset model of spinocerebellar ataxia 3 (SCA3), a polyglutamine disease, which stably expresses expanded CAG repeats in ATXN3. All five offspring of the founder marmoset harbored the transgene with reduced transgene integration sites and without repeat instability or genetic mosaicism, offering improved construct validity. Three of the five marmosets developed progressive motor impairments that segregated into two distinct phenotypes: early onset with rapid progression and late onset with mild progression, accompanied by corresponding patterns in body weight gain and grip strength. Pathological analysis revealed cerebellar Purkinje cell loss, spinal cord neurodegeneration, and widespread intranuclear inclusions. The severity of motor phenotypes correlated with transgene expression levels in disease-relevant brain regions, including the cerebellum, spinal cord, and striatum. By overcoming the common translational limitations of rodent systems, our second-generation model offers a powerful platform for investigating disease mechanisms and testing potential therapeutic interventions. Our results advance the utility of transgenic marmosets as clinically relevant models of neurodegenerative diseases. Summary StatementSecond-generation transgenic marmoset models of spinocerebellar ataxia 3 replicated the progressive motor deficits and neuropathology of the founder marmoset, providing a powerful platform for studying disease mechanisms and developing therapies.

neuroscience↗

bioRxiv2024

Multi-timescale neural adaptation underlying long-term musculoskeletal reorganization.

The central nervous system (CNS) can effectively control body movements despite environmental changes. While much is known about adaptation to external environmental changes, less is known about responses to internal bodily changes. This study investigates how the CNS adapts to long-term alterations in the musculoskeletal system using a tendon transfer model in non-human primates. We surgically relocated finger flexor and extensor muscles to examine how the CNS adapts its strategy for finger movement control by measuring muscle activities during grasping tasks. Two months post-surgery, the monkeys demonstrated significant recovery of grasping function despite the initial disruption. Our findings suggest a two-phase CNS adaptation process: an initial phase enabling function with the transferred muscles, followed by a later phase abandoning this enabled function and restoring a control strategy that, while potentially less conflicted than the maladaptive state, resembled the original pattern, possibly representing a good enough solution. These results highlight a multi-phase CNS adaptation process with distinct time constants in response to sudden bodily changes, offering potential insights into understanding and treating movement disorders. SIGNIFICANCE STATEMENTAfter major changes to the bodys mechanics, the nervous system adapts using strategies on multiple timescales. Our primate tendon transfer study shows that core muscle synergy groupings remain stable, reflecting a default to modular control. However, the activation of these synergies changes dramatically; an initial, rapid swap of their timing proves to be maladaptive, impairing motor function. This conflict is only resolved through the gradual development of slower, compensatory strategies over several weeks. This process highlights the fundamental tension the CNS faces when its reliance on stable motor modules conflicts with the need for flexible control, offering insights into neural plasticity and staged rehabilitation.

neuroscience↗

bioRxiv2024

Spinal and cortical premotor control of primate dexterous hand movements

Skilled hand movements are an evolutionary advance in primates. Phylogenetically distinct corticospinal pathways are involved in hand control: "newer" direct corticomotoneuronal (CM) pathways and "older" indirect corticospinal pathways mediated by spinal premotor interneurons (PreM-INs). However, the functional differences of these pathways remain unclear. Here we show that CM cells and PreM-INs have distinct physiological properties for the activation of hand muscles and provide different types of movement control signals while monkeys perform a precision grip task. Spike-triggered averaging of electromyographic activity indicated that PreM-INs coactivate a larger number of muscles, whereas CM cells more selectively control fewer muscles. The firing activity of PreM-INs was tightly correlated with their target muscle activity and had a greater contribution to generating hand muscle activity. In contrast, CM cell activity diverged temporally from the target muscle activity and had a smaller contribution to its generation. On the basis of these results, we hypothesize that PreM-INs produce gross muscle activity by activating synergistic muscles, whereas CM cells fine-tune target muscle activity. This idea was supported by dimensional reduction analyses of hand muscle activity, as PreM-IN activity was specifically correlated with lower dimensional control of muscle activity, and CM cell activity was correlated with higher dimensional control. These results indicate that the two pathways have distinct functions, synergistic control and fine tuning of hand muscle activity, both of which are essential for the development of dexterous hand movement in primates.

neuroscience↗

bioRxiv2024

Bidirectional Optogenetic Modulations of Peripheral Sensory Nerve Activity: Induction vs. Suppression through Channelrhodopsin and Halorhodopsin

In this study, we investigated the potential of optogenetics for modulating activity of peripheral sensory nerves, particularly tactile and proprioceptive afferents, which are vital for movement control. Using adeno-associated virus serotype 9 vector, we selectively transduced channelrhodopsin (ChR2) and halorhodopsin (eNpHR3.0) into large-diameter sciatic nerve afferents of rats. Diverging from conventional dorsal root ganglion (DRG) approaches, we applied optical stimulation at the distal portion of the afferent nerve. The intensity of optical stimulation varied to modulate the extent of induction and suppression of afferent activity. Then, the effect of optical stimulation was determined by the activity recorded in the dorsal root of the same afferents. Our findings show successful induction and suppression of activity in large-diameter afferents via optical stimulation. By increasing the intensity of blue (for ChR2) and yellow (for eNpHR3.0) light stimulation, the activity of fast-conducting afferent fibers was preferentially evoked or inhibited in an intensity-dependent manner. These data indicate that the activity of large-diameter afferents can systematically be regulated by optogenetics. The present innovative methodology for manipulating specific sensory modalities at the nerve level offers a targeted and accessible alternative to DRG stimulation, expanding the therapeutic scope of optogenetics for treating sensory disorders.

neuroscience↗

bioRxiv2024

Developing, characterizing and modelingCRISPR-based point-of-use pathogen diagnostics

Recent years have seen intense interest in the development of point-of-care nucleic acid diagnostic technologies to address the scaling limitations of laboratory-based approaches. Chief among these are combinations of isothermal amplification approaches with CRISPR-based detection and readouts of target products. Here, we contribute to the growing body of rapid, programmable point-of-care pathogen tests by developing and optimizing a one-pot NASBA-Cas13a nucleic acid detection assay. This test uses the isothermal amplification technique NASBA to amplify target viral nucleic acids, followed by Cas13a-based detection of amplified sequences. We first demonstrate an in-house formulation of NASBA that enables optimization of individual NASBA components. We then present design rules for NASBA primer sets and LbuCas13a guide RNAs for fast and sensitive detection of SARS-CoV-2 viral RNA fragments, resulting in 20 - 200 aM sensitivity without any specialized equipment. Finally, we explore the combination of high-throughput assay condition screening with mechanistic ordinary differential equation modeling of the reaction scheme to gain a deeper understanding of the NASBA-Cas13a system. This work presents a framework for developing a mechanistic understanding of reaction performance and optimization that uses both experiments and modeling, which we anticipate will be useful in developing future nucleic acid detection technologies.

synthetic biology↗

bioRxiv2023

Structure-conditioned masked language models for protein sequence design generalize beyond the native sequence space

Machine learning has revolutionized computational protein design, enabling significant progress in protein backbone generation and sequence design. Here, we introduce Frame2seq, a structure-conditioned masked language model for protein sequence design. Frame2seq generates sequences in a single pass, achieves 49.1% sequence recovery on the CATH 4.2 test dataset, and accurately estimates the error in its own predictions, outperforming the autoregressive ProteinMPNN model with over six times faster inference. To probe the ability of Frame2seq to generate novel designs beyond the native-like sequence space it was trained on, we experimentally test 26 Frame2seq designs for de novo backbones with low identity to the starting sequences. We show that Frame2seq successfully designs soluble (22/26), monomeric, folded, and stable proteins (17/26), including a design with 0% sequence identity to native. The speed and accuracy of Frame2seq will accelerate exploration of novel sequence space across diverse design tasks, including challenging applications such as multi-objective optimization.

bioengineering↗

bioRxiv2023

An intronic copy number variation in Syntaxin 17 determines speed of greying and melanoma incidence in Grey horses

The Greying with age phenotype involves loss of hair pigmentation whereas skin pigmentation is not reduced and a predisposition to melanoma. The causal mutation was initially reported as a duplication of a 4.6 kb intronic sequence in Syntaxin 17. The speed of greying varies considerably among Grey horses. Here we demonstrate the presence of two different Grey alleles, G2 carrying two tandem copies of the duplicated sequence and G3 carrying three. The latter is by far the most common allele, probably due to strong selection for the striking white phenotype. Our results reveal a remarkable dosage effect where the G3 allele is associated with fast greying and high incidence of melanoma whereas G2 is associated with slow greying and low incidence of melanoma. Epigenetic analysis, based on nanopore sequencing of genomic DNA, reveals a drastic reduction in DNA methylation in part of the duplicated sequence harboring MITF binding sites. The copy number expansion transforms a weak enhancer to a strong melanocyte-specific enhancer that underlies hair greying (G2 and G3) and a drastically elevated risk of melanoma (G3 only).

genetics↗