Name:- Ranjana M

Regi. No.: 200810186598

Enrollment no:20/72/BA/001
M.Sc. Ayurveda Biology

Introduction to Methylation:
Methylation is a fundamental biochemical process that plays a critical role in regulating gene
expression, maintaining genomic stability, and modulating cellular function. It involves the
addition of a methyl group (-CH3) to a substrate molecule, which can include DNA, RNA,
proteins, lipids, and small molecules. This covalent modification alters the properties of the
substrate and can influence its structure, stability, and interactions with other molecules. In the
context of DNA methylation, the addition of a methyl group occurs primarily at cytosine bases
within the DNA sequence, forming 5-methylcytosine (5mC). This modification predominantly
occurs at cytosine-guanine dinucleotides (CpG sites), although non-CpG methylation has been
observed in specific cell types, particularly during early development and in neuronal tissues.
DNA methylation is catalyzed by a family of enzymes known as DNA methyltransferases
(DNMTs), which transfer a methyl group from the methyl donor molecule S-adenosyl
methionine (SAM) to the carbon-5 position of cytosine. DNMTs play essential roles in
establishing and maintaining DNA methylation patterns throughout development and adulthood.
In addition to DNMTs, active DNA demethylation processes mediated by enzymes such as
ten-eleven translocation (TET) proteins contribute to the dynamic regulation of DNA
methylation patterns.

Histone methylation involves the addition of methyl groups to specific lysine or arginine
residues on histone proteins, which are the building blocks of chromatin. This modification can
occur at multiple sites across the histone tails and can result in either transcriptional activation or
repression, depending on the specific histone residue and the degree of methylation. Histone
methyltransferases (HMTs) catalyze the transfer of methyl groups to histone substrates, while
histone demethylases (HDMs) remove methyl groups, allowing for the dynamic regulation of
histone methylation patterns. Beyond DNA and histone methylation, methylation also occurs in
RNA molecules, where it can regulate RNA stability, splicing, and translation. Additionally,
methylation of proteins, lipids, and small molecules has diverse functional consequences,
including modulation of enzymatic activity, signaling pathways, and cellular localization.

Overall, methylation is a highly dynamic and regulated process that plays essential roles in
numerous biological processes, including gene regulation, development, differentiation, and
disease pathogenesis. Understanding the mechanisms and functional consequences of
methylation is crucial for elucidating the complexities of cellular biology and for developing
therapeutic interventions targeting epigenetic dysregulation in human health and disease.
Methylation Mechanisms:
Methylation mechanisms involve a series of enzymatic reactions that result in the addition of
methyl groups to specific molecules. These reactions are catalyzed by a diverse array of
methyltransferase enzymes, each with unique substrate specificity and regulatory mechanisms.

In DNA methylation, the primary enzymes responsible for adding methyl groups to cytosine
residues are DNMTs. DNMTs utilize SAM as a methyl donor molecule and catalyze the transfer
of a methyl group to the carbon-5 position of cytosine, forming 5mC. DNMT1 is responsible for
maintaining DNA methylation patterns during DNA replication, while DNMT3A and DNMT3B
are primarily involved in de novo methylation during early development and cellular
differentiation. In addition to these canonical DNMTs, recent studies have identified novel DNA
methyltransferases, such as DNMT3C and DNMT3L, which exhibit unique expression patterns
and functional roles in specific cell types and contexts.

Active DNA demethylation processes counterbalance DNA methylation by removing methyl

groups from 5mC bases. The TET family of enzymes, including TET1, TET2, and TET3,
oxidize 5mC to form 5-hydroxymethylcytosine (5hmC), which serves as an intermediate in the
DNA demethylation pathway. Subsequent enzymatic reactions mediated by TET proteins and
thymine DNA glycosylase (TDG) result in the replacement of 5mC with an unmodified cytosine
base. Histone methylation is catalyzed by histone methyltransferases (HMTs), which transfer
methyl groups from SAM to specific lysine or arginine residues on histone proteins. These
modifications can occur on multiple histone tails, including histone H3 (e.g., H3K4, H3K9,
H3K27) and histone H4 (e.g., H4K20). The functional consequences of histone methylation
depend on the specific histone residue and the degree of methylation. For example,
trimethylation of H3K4 and H3K36 is associated with transcriptional activation, whereas
trimethylation of H3K9 and H3K27 is associated with transcriptional repression.

In addition to DNA and histone methylation, methylation of RNA molecules is catalyzed by

RNA methyltransferases, which add methyl groups to specific nucleotide residues, such as
adenosine (m6A), cytosine (m5C), and guanosine (m7G). These RNA modifications play critical
roles in regulating RNA stability, splicing, translation, and other aspects of RNA metabolism.
Overall, methylation mechanisms are highly dynamic and tightly regulated processes that
contribute to the epigenetic regulation of gene expression, chromatin structure, and cellular
function. Dysregulation of methylation pathways has been implicated in various human diseases,
including cancer, neurological disorders, and developmental syndromes, highlighting the
importance of understanding the molecular mechanisms underlying methylation dynamics.

Regulation of Gene Expression:

The regulation of gene expression is a complex and tightly controlled process that involves
multiple layers of epigenetic regulation, including DNA methylation, histone modifications,
chromatin remodeling, and non-coding RNAs. Methylation, particularly DNA and histone
methylation, plays a central role in modulating gene expression patterns by influencing
chromatin structure, DNA accessibility, and the recruitment of transcriptional regulatory
complexes.

In DNA methylation, the addition of methyl groups to cytosine residues within gene regulatory
regions, such as promoters and enhancers, can inhibit the binding of transcription factors and
recruit proteins involved in gene silencing. This results in the formation of condensed chromatin
structures (heterochromatin) and transcriptional repression of the associated gene. DNA
methylation-mediated gene silencing is essential for the maintenance of cell identity and
tissue-specific gene expression patterns during development and differentiation. Conversely,
active DNA demethylation processes, mediated by enzymes such as ten-eleven translocation
(TET) proteins, can remove methyl groups from DNA, leading to gene activation and
transcriptional derepression.

Histone methylation also contributes to the regulation of gene expression by modulating

chromatin structure and recruiting chromatin-modifying complexes. Histone methyltransferases
(HMTs) catalyze the addition of methyl groups to specific lysine or arginine residues on histone
tails, while histone demethylases (HDMs) remove methyl groups, allowing for the dynamic
regulation of histone methylation patterns. The functional consequences of histone methylation
depend on the specific histone residue and the degree of methylation. For example,
trimethylation of histone H3 lysine 4 (H3K4me3) is associated with transcriptional activation,
whereas trimethylation of histone H3 lysine 9 (H3K9me3) is associated with transcriptional
repression. In addition to direct effects on chromatin structure and transcriptional regulation,
DNA and histone methylation can also influence gene expression indirectly by modulating the
recruitment of chromatin-modifying complexes, non-coding RNAs, and transcriptional
regulatory factors. For example, methyl-CpG binding domain proteins (MBDs) can recognize
and bind to methylated CpG sites, leading to the recruitment of histone deacetylases (HDACs)
and other chromatin-modifying enzymes that promote gene silencing.

Furthermore, DNA and histone methylation can interact with other epigenetic modifications,
such as histone acetylation and DNA hydroxymethylation, to fine-tune gene expression patterns
and cellular responses to internal and external stimuli. Crosstalk between different epigenetic
pathways adds another layer of complexity to the regulation of gene expression and underscores
the importance of integrative approaches in understanding epigenetic regulation in health and
disease. Overall, methylation-mediated regulation of gene expression is a dynamic and highly
orchestrated process that plays critical roles in development, differentiation, cellular homeostasis,
and disease pathogenesis. Elucidating the molecular mechanisms underlying methylation
dynamics and its functional consequences on gene expression will provide valuable insights into
the complexity of epigenetic regulation and may lead to the development of novel therapeutic
strategies for treating human diseases.

Methylation in Health and Disease:

Methylation patterns are dynamically regulated and can be influenced by various factors,
including age, environmental exposures, genetic variation, and lifestyle choices. Aberrant
methylation patterns have been implicated in a wide range of human diseases, including cancer,
neurological disorders, cardiovascular diseases, metabolic disorders, and autoimmune diseases.
Understanding the role of methylation in health and disease is essential for elucidating disease
mechanisms, identifying diagnostic biomarkers, and developing targeted therapeutic
interventions.

In cancer, alterations in DNA methylation patterns are a hallmark feature of tumor cells and
contribute to tumor initiation, progression, metastasis, and treatment resistance. Global
hypomethylation of repetitive DNA sequences and focal hypermethylation of gene promoters are
common epigenetic alterations observed in various cancer types. Hypomethylation of repetitive
DNA elements, such as long interspersed nuclear elements (LINE-1) and Alu sequences, can
lead to genomic instability, chromosomal rearrangements, and activation of oncogenic pathways.
Focal hypermethylation of tumor suppressor gene promoters, such as CDKN2A (p16), MLH1,
and BRCA1, can result in transcriptional silencing and loss of tumor suppressor function.

In addition to DNA methylation changes, alterations in histone modifications, chromatin

remodeling, and non-coding RNA expression contribute to the epigenetic landscape of cancer
cells and influence tumor phenotype and behavior. Dysregulation of histone modifying enzymes,
such as histone methyltransferases (HMTs) and histone demethylases (HDMs), can disrupt
chromatin structure and transcriptional regulation, leading to aberrant gene expression patterns
and malignant transformation. Similarly, aberrant expression of non-coding RNAs, such as
microRNAs (miRNAs) and long non-coding RNAs (lncRNAs), can modulate gene expression
networks and contribute to cancer pathogenesis. In addition to cancer, alterations in methylation
patterns have been implicated in various neurological disorders, including Alzheimer's disease,
Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), schizophrenia,
bipolar disorder, and autism spectrum disorders. Dysregulation of DNA methylation, histone
modifications, and non-coding RNA expression has been observed in postmortem brain tissues
and peripheral blood samples from patients with neurological disorders, suggesting a role for
epigenetic mechanisms in disease pathogenesis.

Furthermore, methylation changes have been linked to cardiovascular diseases, such as

atherosclerosis, hypertension, myocardial infarction, heart failure, and stroke. DNA methylation
alterations in genes involved in lipid metabolism, inflammation, oxidative stress, and vascular
function have been associated with cardiovascular risk factors, disease severity, and prognosis.
Epigenetic modifications, such as histone acetylation and DNA methylation, regulate the
expression of genes involved in endothelial dysfunction, smooth muscle cell proliferation,
macrophage activation, and plaque stability, thereby influencing the development and
progression of atherosclerosis and its complications.

Moreover, methylation patterns have been implicated in metabolic disorders, such as obesity,
type 2 diabetes, and metabolic syndrome. Dysregulation of DNA methylation, histone
modifications, and non-coding RNA expression in adipose tissue, liver, muscle, pancreas, and
brain has been linked to alterations in energy metabolism, insulin sensitivity, glucose
homeostasis, lipid metabolism, and inflammation. Epigenetic modifications can modulate the
expression of genes involved in adipogenesis, lipogenesis, insulin signaling, glucose transport,
mitochondrial function, and inflammatory pathways, contributing to the pathogenesis of
metabolic diseases.

Additionally, methylation changes have been associated with autoimmune diseases, such as
systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), multiple sclerosis (MS), type 1
diabetes (T1D), and inflammatory bowel disease (IBD). Dysregulation of DNA methylation,
histone modifications, and non-coding RNA expression in immune cells and target tissues has
been implicated in the activation of autoreactive immune responses, tissue inflammation, and
organ damage. Epigenetic modifications can modulate the expression of genes involved in
immune cell differentiation, activation, survival, migration, and effector function, thereby
influencing the pathogenesis of autoimmune diseases. Overall, methylation patterns are
dynamically regulated and play critical roles in human health and disease. Aberrant methylation
patterns have been implicated in various diseases across different organ systems and contribute
to disease initiation, progression, and complications. Elucidating the molecular mechanisms
underlying methylation dysregulation in disease states holds promise for identifying novel
diagnostic biomarkers and therapeutic targets, as well as developing personalized treatment
strategies for patients with complex and heterogeneous diseases.

Environmental Influences on Methylation:

Environmental factors, including diet, lifestyle, chemical exposures, psychosocial stress,
socioeconomic status, and physical activity, can influence DNA methylation patterns and
contribute to disease risk and health outcomes. Epigenetic modifications, such as DNA
methylation, histone modifications, and non-coding RNA expression, serve as molecular
mechanisms by which environmental exposures can alter gene expression patterns and cellular
phenotypes. Dietary factors, such as nutrients, bioactive compounds, and dietary patterns, have
been shown to modulate DNA methylation patterns and gene expression profiles. For example,
methyl donors, such as folate, vitamin B12, choline, and methionine, are essential cofactors for
DNA methylation reactions and play critical roles in one-carbon metabolism. Adequate intake of
methyl donor nutrients is associated with global DNA hypomethylation and promoter-specific
DNA hypermethylation, whereas deficiencies or excesses of these nutrients can disrupt DNA
methylation patterns and increase disease risk. Similarly, bioactive compounds found in fruits,
vegetables, whole grains, nuts, seeds, herbs, spices, teas, and other plant-based foods can
influence DNA methylation patterns and gene expression profiles. Phytochemicals, such as
polyphenols, flavonoids, isothiocyanates, allyl sulfides, and lignans, have been shown to
modulate epigenetic regulators, including DNMTs, HMTs, HDMs, and non-coding RNAs. These
dietary compounds can exert chemoprotective effects against cancer, cardiovascular diseases,
neurodegenerative diseases, metabolic disorders, and inflammatory conditions by regulating
epigenetic pathways involved in cell growth, differentiation, apoptosis, angiogenesis,
inflammation, and oxidative stress.

Furthermore, dietary patterns, such as the Mediterranean diet, DASH diet, vegetarian diet, vegan
diet, paleolithic diet, and low-carbohydrate diet, have been associated with distinct DNA
methylation signatures and gene expression profiles. Epidemiological studies have shown that
adherence to healthy dietary patterns is inversely correlated with DNA methylation age
acceleration, a biomarker of biological aging, and is associated with reduced risk of chronic
diseases and increased longevity. Conversely, adherence to unhealthy dietary patterns, such as
the Western diet, high-fat diet, high-sugar diet, processed food diet, and fast food diet, is
positively correlated with DNA methylation age acceleration and is associated with increased
risk of chronic diseases and premature mortality. In addition to diet, lifestyle factors, such as
physical activity, sleep quality, smoking, alcohol consumption, and substance abuse, can impact
DNA methylation patterns and gene expression profiles. Regular exercise has been shown to
promote global DNA hypomethylation and modulate the expression of genes involved in
metabolism, inflammation, oxidative stress, and neuroplasticity. Adequate sleep duration and
quality are associated with favorable DNA methylation patterns and reduced risk of metabolic
diseases, cardiovascular diseases, and cognitive decline. Conversely, chronic sleep deprivation
and poor sleep quality are associated with aberrant DNA methylation patterns and increased risk
of chronic diseases, mental health disorders, and neurodegenerative diseases.

Moreover, tobacco smoke contains numerous toxicants and carcinogens that can alter DNA
methylation patterns and gene expression profiles. Cigarette smoking is associated with global
DNA hypermethylation and site-specific DNA hypomethylation, particularly in genes involved
in cancer development, inflammation, and oxidative stress. Similarly, alcohol consumption can
influence DNA methylation patterns and gene expression profiles in the liver, brain, immune
cells, and other tissues. Chronic alcohol abuse is associated with global DNA hypomethylation
and site-specific DNA hypermethylation, leading to dysregulation of metabolic pathways,
immune responses, and neuronal signaling pathways. Furthermore, psychosocial stressors, such
as socioeconomic disadvantage, childhood adversity, traumatic experiences, social isolation,
work-related stress, and caregiving responsibilities, can impact DNA methylation patterns and
gene expression profiles. Chronic stress exposure is associated with alterations in
stress-responsive genes, inflammatory genes, and neuroendocrine genes, leading to dysregulation
of the hypothalamic-pituitary-adrenal (HPA) axis, sympathetic-adrenal-medullary (SAM) axis,
immune system, and central nervous system. These epigenetic changes may contribute to the
development of stress-related disorders, such as depression, anxiety, post-traumatic stress
disorder (PTSD), and cardiovascular diseases.

Overall, environmental influences on DNA methylation are multifaceted and encompass a wide
range of factors, including diet, lifestyle, chemical exposures, psychosocial stress,
socioeconomic status, and physical activity. These environmental exposures can impact DNA
methylation patterns and gene expression profiles, leading to alterations in cellular phenotypes,
physiological functions, and disease risk. Understanding the complex interplay between
environmental factors and epigenetic regulation is essential for elucidating disease mechanisms,
identifying preventive strategies, and developing personalized interventions for optimizing health
and well-being.

Techniques for Studying Methylation:

The study of methylation encompasses a wide range of techniques and methodologies for
profiling DNA methylation, histone modifications, and RNA modifications in various biological
samples. These techniques provide valuable insights into the epigenetic regulation of gene
expression, chromatin structure, and cellular function in health and disease. One of the most
widely used techniques for studying DNA methylation is bisulfite conversion coupled with DNA
sequencing. Bisulfite treatment chemically converts unmethylated cytosine residues to uracil
while leaving methylated cytosine residues unchanged. Subsequent DNA sequencing or
array-based analysis allows for the quantification of DNA methylation levels at single-base
resolution. Bisulfite sequencing techniques include bisulfite PCR sequencing (BSP),
methylation-specific PCR (MSP), pyrosequencing, and next-generation sequencing (NGS)
methods such as reduced representation bisulfite sequencing (RRBS) and whole-genome
bisulfite sequencing (WGBS). These techniques enable the genome-wide profiling of DNA
methylation patterns and the identification of differentially methylated regions (DMRs)
associated with disease states, developmental stages, and environmental exposures.

Another approach for studying DNA methylation is methylation-sensitive restriction enzyme

digestion coupled with DNA sequencing or array-based analysis. Methylation-sensitive
restriction enzymes, such as HpaII and MspI, cleave DNA at specific recognition sites only if the
cytosine residue is unmethylated. Differential digestion of methylated and unmethylated DNA
fragments allows for the identification of methylation-sensitive restriction sites and the
quantification of DNA methylation levels. Techniques based on methylation-sensitive restriction
enzymes include methylation-sensitive amplified polymorphism (MSAP), restriction landmark
genomic scanning (RLGS), and methylation-specific restriction enzyme digestion followed by
microarray hybridization or sequencing.
In addition to bisulfite conversion and restriction enzyme digestion, antibody-based techniques,
such as methylated DNA immunoprecipitation (MeDIP) and chromatin immunoprecipitation
(ChIP), are commonly used for studying DNA methylation and histone modifications,
respectively. MeDIP utilizes antibodies specific for methylated cytosine residues to enrich
methylated DNA fragments, which can then be analyzed by DNA sequencing, microarray
hybridization, or quantitative PCR. ChIP utilizes antibodies specific for histone modifications to
immunoprecipitate chromatin fragments containing modified histone proteins, followed by DNA
sequencing, microarray hybridization, or quantitative PCR analysis. These techniques allow for
the genome-wide profiling of DNA methylation and histone modification patterns in chromatin
regions associated with gene regulatory elements, such as promoters, enhancers, and insulators.

Furthermore, emerging technologies, such as single-cell epigenomics, long-read sequencing, and

nanopore sequencing, are revolutionizing the field of methylation research by enabling
high-resolution, high-throughput, and single-molecule analysis of DNA methylation patterns in
individual cells and complex tissues. Single-cell epigenomics techniques, such as single-cell
bisulfite sequencing (scBS-seq) and single-cell ATAC-seq (scATAC-seq), allow for the
characterization of DNA methylation and chromatin accessibility profiles in heterogeneous cell
populations, providing insights into cellular heterogeneity, lineage commitment, and
developmental trajectories. Long-read sequencing technologies, such as Pacific Biosciences
(PacBio) and Oxford Nanopore Technologies (ONT), enable the direct detection of DNA
modifications, such as methylation, hydroxymethylation, and other base modifications, at
single-base resolution without the need for bisulfite conversion or restriction enzyme digestion.
These technologies offer advantages for studying DNA methylation patterns in repetitive
regions, structural variants, and complex genomic loci that are challenging to analyze using
short-read sequencing platforms.

Overall, the study of methylation relies on a diverse array of techniques and methodologies for
profiling DNA methylation, histone modifications, and RNA modifications in various biological
samples. These techniques provide valuable insights into the epigenetic regulation of gene
expression, chromatin structure, and cellular function in health and disease, and enable the
identification of biomarkers, therapeutic targets, and personalized interventions for optimizing
human health and well-being
Methods to detect DNA methylation

Bisulfite Conversion Combined with PCR (BSP/MSP):

Principle: This method relies on the chemical conversion of unmethylated cytosine residues to
uracil through treatment with sodium bisulfite. Methylated cytosines are resistant to this
conversion. Following bisulfite treatment, PCR amplification is performed using primers specific
to the region of interest, incorporating the converted DNA. Subsequent sequencing or analysis
distinguishes methylated from unmethylated cytosines.
Applications: BSP and MSP are commonly employed to assess DNA methylation at specific
loci, particularly CpG islands within gene promoters. BSP provides quantitative information
about the methylation status of individual CpG sites, while MSP offers a qualitative assessment
by targeting either methylated or unmethylated sequences. These methods are valuable for
studying targeted regions of interest and investigating DNA methylation changes associated with
gene expression regulation, developmental processes, and disease states.

Bisulfite Sequencing (Whole-Genome Bisulfite Sequencing - WGBS):

Principle: WGBS begins with bisulfite treatment of genomic DNA, converting unmethylated
cytosines to uracil while preserving methylated cytosines. Following library preparation,
next-generation sequencing (NGS) is performed to generate sequence data. Computational
analysis then compares the sequenced reads to a reference genome, identifying methylated and
unmethylated cytosines at single-base resolution.
Applications: WGBS enables genome-wide profiling of DNA methylation patterns across all
CpG sites in the genome. By providing comprehensive coverage and high resolution, this method
allows researchers to identify differentially methylated regions (DMRs) associated with
biological processes, such as development, aging, and disease. WGBS is particularly valuable for
investigating global changes in DNA methylation and understanding the regulatory mechanisms
underlying epigenetic modifications.

Methylated DNA Immunoprecipitation (MeDIP):

Principle: MeDIP involves the enrichment of methylated DNA fragments using antibodies
specific to 5-methylcytosine (5mC). Following immunoprecipitation, the enriched DNA is
isolated and subjected to downstream analysis, such as microarray hybridization or
next-generation sequencing (MeDIP-seq), to identify methylated regions across the genome.
Applications: MeDIP allows for the genome-wide profiling of DNA methylation patterns,
focusing on regions enriched for methylated cytosines. This method is particularly useful for
identifying DNA methylation changes associated with regulatory elements, such as promoters,
enhancers, and CpG islands. MeDIP-seq data provide insights into the distribution and
abundance of DNA methylation marks, aiding in the discovery of epigenetic biomarkers and the
characterization of disease-related epigenetic alterations.

Chromatin Immunoprecipitation (ChIP) Combined with Bisulfite Sequencing

(ChIP-BS-seq):

Principle: ChIP captures chromatin fragments containing specific histone modifications or

DNA-binding proteins using antibodies. Following immunoprecipitation, the captured chromatin
is subjected to bisulfite treatment and sequencing to analyze DNA methylation patterns within
the targeted chromatin regions.
Applications: ChIP-BS-seq integrates epigenetic data from histone modifications or
DNA-binding proteins with DNA methylation information, allowing researchers to investigate
the interplay between chromatin structure and DNA methylation. This method provides insights
into the regulatory mechanisms governing epigenetic modifications and their impact on gene
expression regulation, cellular differentiation, and disease development.

Enhanced Reduced Representation Bisulfite Sequencing (ERRBS):

Principle: ERRBS combines reduced representation bisulfite sequencing (RRBS) with

additional steps to enrich for CpG-rich regions of the genome. This approach focuses on
sequencing regions with high CpG density, such as gene promoters and CpG islands, providing
enhanced coverage and resolution compared to traditional bisulfite sequencing methods.
Applications: ERRBS is particularly useful for studying DNA methylation patterns in CpG-rich
regions associated with gene regulatory elements. By targeting biologically relevant CpG sites,
ERRBS enables the identification of DNA methylation changes involved in gene regulation,
cellular differentiation, and disease processes. This method is valuable for characterizing
epigenetic alterations associated with specific genomic loci and elucidating their functional
consequences.

Nanopore Sequencing:

Principle: Nanopore sequencing directly detects changes in DNA sequence and modifications,
such as DNA methylation, by measuring alterations in electrical current as DNA molecules pass
through a nanopore. This approach provides real-time sequencing data and can detect modified
bases without the need for chemical conversion.
Applications: Nanopore sequencing offers long-read capabilities and real-time analysis, making
it suitable for studying DNA methylation patterns in complex genomic regions and
heterogeneous samples. This method enables the detection of DNA modifications at single-base
resolution, providing insights into the dynamics of epigenetic modifications and their functional
consequences in gene regulation, genome stability, and disease development.
These methods offer diverse strategies for studying DNA methylation patterns and their
regulatory roles in biological processes. By leveraging these techniques, researchers can gain
comprehensive insights into the dynamic nature of epigenetic modifications and their impact on
gene expression, cellular function, and disease states.