⚡ Epigenetics Crash Course

8 Topics · Flashcards · Diagrams · Memory Tricks — You've got this!

1 Epigenetics: Overview & Definitions
Epigenetics = Heritable changes in cellular function or gene expression transmitted cell-to-cell (or generation-to-generation) via chromatin-based molecular signals — without altering the DNA sequence.
Epigenomics = Genome-wide study of epigenetic patterns (methylome, histone landscapes, nucleosome positioning across cell types).
🧠 Memory Trick
"Epi = Above" — Think of it as notes written on top of the DNA without changing the letters. Like sticky notes on a textbook — the book stays the same, but the notes change how you read it!

Two Key Properties

Dynamic & Transient
Rapid response to cell needs
OR
🔒
Long-lasting
Transmitted through cell divisions / generations

The Big 3 Mechanisms

EPIGENETIC MECHANISMS DNA Methylation → Repression Histone Modifications → Activation OR Repression Histone Variants
Fig: The three major epigenetic mechanisms (Fig. 3-8)
🧠 Mnemonic: "MoMoVa"
Methylation (DNA) · Modifications (Histones) · Variants (Histones)
Key distinction from genetics: epigenetic marks are sequence-independent
Clinical Environmental/lifestyle factors alter epigenetic marks → disease. Reversibility = therapeutic opportunity. Large-scale projects: ENCODE, methylome catalogues.
2 DNA Methylation
Methylation of cytosine C5 (carbon 5 of the pyrimidine ring) → produces 5-methylcytosine (5-mC). Occurs almost exclusively at CpG dinucleotides.
Methyl-CpG binding proteins
Recruit chromatin-modifying enzymes
SILENCE transcription

5-mC vs 5-hmC

5-mC
(methylcytosine)
Stable mark, inherited through cell division
~5% of all cytosines
vs
5-hmC
(hydroxymethylcytosine)
Variable; highest in regulatory regions
0.1–1% of cytosines
Likely demethylation intermediate
5-mC enzymatic 5-hmC further steps Unmethylated C Demethylation pathway (Fig. 3-9)
🧠 Memory Trick
"Methylation = Mute" — Both start with M. Methylate a gene → mute/silence it!
"CpG = C-phosphate-G" — The p is the phosphodiester bond between C and G.
Demethylation: 5-mC → 5-hmC (enzymatic). Extensive demethylation during germ cell development & early embryogenesis (restores totipotency/pluripotency).
Clinical Cancer: hypomethylation of large genomic segments OR regional hypermethylation at CpG islands (Ch. 15). Differential methylation at specific loci + sequence variation → modulates genetic risk (e.g., rheumatoid arthritis).
3 Histone Modifications & Variants
Modifications occur on N-terminal "tails" of core histones H2A, H2B, H3, H4, extending from the nucleosome core (Fig. 3-8).

Key Modification Types

Methylation
Acetylation
Phosphorylation

Must-Know Marks

MarkExampleEffect
H3K9 methylation H3K9me Repressive — silent chromatin
H3K27 acetylation H3K27ac Activating — open regulatory regions
🧠 Memory Trick
"K9 silences like a K-9 police dog guarding the gene" — H3K9me = repressive!
"Acetylation = Activation" — Both start with "A"! H3K27ac = active.
NUCLEOSOME H2A H2B H3 H4 DNA Me H3K9 Ac H3K27 P Phos
Histone tails protruding from nucleosome with modifications
ENCODE Project: identified 12 common modifications across ~50 cell types, attributing putative function to >50% of the genome — far more than the ~2% that is "coding."

Histone Variants

Encoded by separate genes; replace core histones to generate specialized chromatin.

VariantLocationFunction
CENP-ACentromeres onlyMarks kinetochore location
H2A.XSites of DNA damageMarks DSBs requiring repair
🧠 Memory Trick
"CENP-A = CENter Piece" — sits at the center (centromere) of the chromosome.
"H2A.X = X marks the spot" — X marks the damage!
4 Chromatin Architecture & 3D Genome
The genome is NOT a linear string — it adopts a highly ordered 3D arrangement in the nucleus, guided by epigenetic signals. The 3D landscape predicts the transcriptome of any cell type.

Four Levels of Organization

LevelFeatureSignificance
Chromosome territories Each chromosome occupies a distinct nuclear territory Proximity influences abnormality incidence
Subchromosomal domains Megabase-sized; GC/AT rich, gene density, replication timing Coordinate gene expression at chromosome level
Chromatin loops Bring enhancers/locus-control regions to promoters Long-range gene activation
Nucleosome positioning Gaps between nucleosomes expose DNA TF and regulatory protein binding
NUCLEUS Chr 1 territory Chr 7 territory Chr 22 territory chromatin loop enhancer promoter
Chromosome territories and chromatin loops within the nucleus (Fig. 3-10)
🧠 Memory Trick — "TSLN" (Top → Small)
Territories → Subchromosomal domains → Loops → Nucleosome positioning
Think: "This Structure Lets Nature" organize genes in 3D
5 Allelic Imbalance in Gene Expression
Genes present in two copies are not always expressed equally from both alleles. Three major patterns:
Balanced Biallelic G A Equal RNA from both Allelic Imbalance G > A Unequal RNA; one dominates Monoallelic A only Only one allele expressed
Three patterns of allelic expression (Fig. 3-11)
Prevalence: 5–20% of autosomal genes show unequal expression; usually <2-fold, but up to 10-fold differences seen.
Cause: Sequence variants alter TF binding or DNA methylation patterns at the two alleles.
🧠 Memory Trick
Think of it as a volume knob: Balanced = both speakers equal, Imbalanced = one speaker louder, Monoallelic = one speaker completely off.
6 Monoallelic Expression: 4 Mechanisms
Four distinct ways a cell can express only one of its two alleles (Table 3-2):
TypeGenesBasisOrigin
Somatic rearrangement Immunoglobulins, T-cell receptors DNA cutting & pasting → functional gene on one allele only B- & T-cell lineages
Random monoallelic silencing Olfactory receptors; up to 10% of genes in some cell types Differential epigenetic packaging at locus Specific cell types
Genomic imprinting >100 developmental genes Imprinted region epigenetically marked by parent of origin in germline Parental germline
X chromosome inactivation Most X-linked genes in females Epigenetic silencing of one entire X chromosome Early embryogenesis
4 Mechanisms of Monoallelic Expression SOMATIC REARRANGEMENT DNA cutting/pasting Ig + TCR genes B & T cells only RANDOM SILENCING Epigenetic packaging Olfactory receptors Up to 10% of genes GENOMIC IMPRINTING Parent-of-origin mark >100 genes Set in germline X CHROMOSOME INACTIVATION Whole X silenced Most X-linked genes ♀ Early embryogenesis DNA-level change ← All three are EPIGENETIC (no DNA sequence change) → 🔪 Rearranges DNA 👨‍👩‍👧 Non-random (parental) 🎲 Random per cell 🎲 Random per cell
Comparison of four monoallelic expression mechanisms (Table 3-2)
🧠 Mnemonic: "SRIX" (like "S-R-I-X")
Somatic rearrangement · Random silencing · Imprinting · X inactivation
Only "S" changes DNA. The rest are epigenetic!
Key distinctions:
  • Somatic rearrangement: UNIQUE — only in B & T cells, rearranges actual DNA. Rearranges hundreds of kilobases. Mature Ig/TCR mRNA is exclusively monoallelic.
  • Random monoallelic: broadens diversity of cellular responses (e.g., 1 OR gene per olfactory sensory neuron from among hundreds of family members). Also seen in immune/chemosensory genes.
7 Genomic Imprinting
Imprinting = Monoallelic expression determined nonrandomly by parental origin. Epigenetic marks established in the germline of one parent.

Core Features

Imprinting Life Cycle ① GAMETOGENESIS Imprint established in one parent's germline ② FERTILIZATION Imprint carried into zygote ③ SOMATIC Maintained through cell divisions ④ RESET Erased & re-established Reversible — must reset in germline for next generation
The imprinting cycle: set → maintain → reset
🧠 Memory Trick
"Imprint = Stamp from parent" — Like a wax seal on a letter. Mom stamps some genes, Dad stamps others. The seal is removed and re-stamped each generation.

vs Random monoallelic: Imprinting is NOT random — it's always the same parent's allele silenced. Like a rule vs a coin flip.
Clinical ~100 imprinted genes on autosomes. Classic disorders:
Prader-Willi syndrome (Case 38) & Beckwith-Wiedemann syndrome (Case 6) — Ch. 6, 7.
8 X Chromosome Inactivation
Most extensive example of random monoallelic expression. Mechanism of dosage compensation in females — silences most genes on one X chromosome.

Key Steps

Early embryogenesis
(first week)
Random choice
per cell
Clonally maintained
all daughter cells
Female = clonal mosaic
X Inactivation → Clonal Mosaic Xm Xp Early embryo cell RANDOM Xm Xp Maternal X active (pink cells) Xm Xp Paternal X active (blue cells) XIST Gene — The Master Switch Located in X inactivation center · Produces long ncRNA Expressed ONLY from the INACTIVE X · Silenced on the active X No XIST = No X inactivation!
X inactivation creates a clonal mosaic in females (Fig. 3-13)

Epigenetic Features of Inactive X

DNA methylation
Histone modifications
macroH2A variant
Inactive X = Barr body (heterochromatic mass visible in interphase nuclei).

Exceptions!

🧠 Memory Tricks
"XIST = eXit sign for X" — XIST RNA coats the X and tells it to shut down (exit)!
"Barr body = Barred from expression" — The Barr body is the inactive X, barred from being read.
"Calico cats" — Classic example of X-inactivation mosaic. Orange/black patches = different X active in each clone!
Clinical Carrier females for X-linked disease show variable phenotype depending on the ratio of Xi vs Xa cells (Ch. 6, 7). Skewed inactivation can cause symptomatic carriers.

🃏 Flashcard Deck

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Topic 1: Overview

What is epigenetics?
Heritable changes in cellular function or gene expression via chromatin-based molecular signals WITHOUT altering the DNA sequence. Transmitted cell-to-cell or across generations.
Name the 3 major epigenetic mechanisms
1) DNA methylation (repression)
2) Histone modifications (activation or repression)
3) Histone variants (mark specific regions)
Two key properties of epigenetic changes?
1) Dynamic & transient (rapid response)
2) Long-lasting (transmitted through cell divisions or generations)
What is epigenomics?
Genome-wide study of epigenetic patterns: methylome, histone landscapes, nucleosome positioning across cell types.

Topic 2: DNA Methylation

Where does DNA methylation occur?
Cytosine C5 (carbon 5 of the pyrimidine ring) → 5-methylcytosine (5-mC). Almost exclusively at CpG dinucleotides.
What is the effect of DNA methylation?
Gene REPRESSION: methyl-CpG binding proteins → recruit chromatin-modifying enzymes → silence transcription. "Methylation = Mute"
5-mC vs 5-hmC: what's the difference?
5-mC: stable mark, ~5% of all cytosines in adults, inherited through division.
5-hmC: variable, 0.1-1% of cytosines, highest in regulatory regions. Likely demethylation intermediate.
When does extensive demethylation occur?
During germ cell development & early embryogenesis — restores totipotency/pluripotency. 5-mC → 5-hmC → unmethylated C.
DNA methylation changes in cancer?
Hypomethylation of large genomic segments OR regional hypermethylation at CpG islands. Both common patterns.

Topic 3: Histone Modifications & Variants

Where do histone modifications occur?
On N-terminal "tails" of core histones H2A, H2B, H3, H4 extending from the nucleosome core.
H3K9me — activating or repressive?
REPRESSIVE — marks silent chromatin.
🧠 "K-9 police dog guards (silences) the gene"
H3K27ac — activating or repressive?
ACTIVATING — marks open regulatory regions.
🧠 "Acetylation = Activation" (both start with A)
What is CENP-A?
H3-related histone VARIANT found only at centromeres. Marks kinetochore location.
🧠 "CENP-A = CENter Piece"
What is H2A.X?
Histone variant that marks sites of DNA double-strand breaks requiring repair.
🧠 "X marks the (damage) spot"
ENCODE Project finding about histone modifications?
12 common modifications across ~50 cell types → putative function for >50% of the genome (far more than ~2% that codes for protein).

Topic 4: 3D Genome

Name the 4 levels of chromatin organization (big → small)
1) Chromosome territories
2) Subchromosomal domains
3) Chromatin loops
4) Nucleosome positioning
🧠 "TSLN — This Structure Lets Nature"
What do chromatin loops accomplish?
Bring enhancers/locus-control regions physically close to promoters → enable long-range gene activation.
Why is nucleosome positioning important?
Gaps between nucleosomes expose DNA → allow TF and regulatory protein binding.

Topic 5: Allelic Imbalance

What are the 3 patterns of allelic expression?
1) Balanced biallelic (G ≈ A)
2) Allelic imbalance (G > A)
3) Monoallelic (A only)
What % of autosomal genes show unequal expression?
5–20% show unequal expression; usually <2-fold difference, but up to 10-fold possible.
What causes allelic imbalance?
Sequence variants that alter TF binding or DNA methylation patterns at the two alleles.

Topic 6: Monoallelic Expression

Name the 4 mechanisms of monoallelic expression
SRIX:
1) Somatic rearrangement (DNA-level)
2) Random monoallelic silencing
3) Imprinting
4) X inactivation
(Only #1 changes DNA; rest are epigenetic)
Which monoallelic mechanism actually changes DNA?
ONLY somatic rearrangement (Ig & TCR genes in B & T cells). The other three are purely epigenetic.
Random monoallelic silencing — classic example?
Olfactory receptors — one OR gene expressed per olfactory sensory neuron from among hundreds of family members. Also seen in immune/chemosensory genes. Up to 10% of genes in some cell types.

Topic 7: Genomic Imprinting

What is genomic imprinting?
Monoallelic expression determined NONRANDOMLY by parental origin. Epigenetic marks set in the germline of one parent.
When are imprints established?
During GAMETOGENESIS (before fertilization). Maintained through somatic cell divisions. Must be REVERSIBLE each generation.
What controls imprinting at a locus?
Imprinting control regions (imprinting centers) — often involve ncRNAs that initiate epigenetic changes spreading outward along the chromosome.
Name 2 clinical imprinting disorders
1) Prader-Willi syndrome (Case 38)
2) Beckwith-Wiedemann syndrome (Case 6)
~100 imprinted genes on autosomes.
How does imprinting differ from random monoallelic silencing?
Imprinting: ALWAYS same parent's allele silenced (nonrandom, set in germline).
Random: coin flip per cell, determined somatically.

Topic 8: X Inactivation

What is X inactivation and its purpose?
Silencing of most genes on one X chromosome in females for DOSAGE COMPENSATION. Most extensive example of random monoallelic expression.
When does X inactivation occur?
Early embryogenesis (first week). Random choice per cell, then clonally maintained → females = clonal mosaic.
What is XIST?
Long ncRNA in the X inactivation center. Expressed ONLY from the INACTIVE X allele; silenced on active X. No XIST = no X inactivation.
What is a Barr body?
The inactive X chromosome visible as a heterochromatic mass in interphase nuclei.
🧠 "Barr = Barred from expression"
Epigenetic features of inactive X? (3 things)
1) DNA methylation
2) Histone modifications
3) Histone variant macroH2A
What % of X-linked genes escape inactivation?
At least 15% show biallelic expression. Also, pseudoautosomal genes (identical on X & Y) do NOT inactivate.

⚡ Quick Reference — Key Comparisons for Exam

ConceptKey DetailMemory Cue
DNA methylationCpG → 5-mC → silenceMethylation = Mute (both M)
H3K9meRepressiveK-9 dog guards the gene
H3K27acActivatingAcetylation = Activation (both A)
CENP-ACentromere variantCENter Piece
H2A.XDNA damage markerX marks the spot
XISTncRNA from inactive XeXIt Sign for X
Barr bodyInactive X in nucleiBarred from expression
ImprintingParent-of-origin, nonrandomWax seal from parent
4 monoallelicS-R-I-XOnly S changes DNA
3D genomeT-S-L-N (big→small)This Structure Lets Nature
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