3.183.193.20(i)3.20(ii)

Controlling Gene Expression

How cells with identical DNA become muscle, nerve, or blood cells — and how expression is regulated at every level.

Every cell in your body contains the same 20,000+ genes. So how does a liver cell ‘know’ to make albumin while a red blood cell makes haemoglobin?
Learning Objectives
AO1 Understand the role of transcription factors in regulating gene expression, including the binding of promoters and enhancers
AO2 Understand post-transcriptional control of gene expression, including alternative RNA splicing
AO1 Understand how epigenetic changes (DNA methylation and histone modification) can modify gene expression
AO2 Understand the role of non-coding RNA in gene regulation, including X-inactivation and Barr bodies

Diagnostic Quiz

Q1
What is the role of RNA polymerase in transcription?
A It joins amino acids together
B It synthesises mRNA from a DNA template
C It unwinds the double helix permanently
D It removes introns from pre-mRNA
Q2
Which of these is NOT part of protein synthesis?
A Transcription
B Methylation
C Translation
D Splicing
Q3
What are introns?
A Coding sequences of a gene
B Non-coding sequences that are removed from pre-mRNA
C Proteins that regulate gene expression
D Sequences that code for tRNA
Productive Struggle

Before you learn the content, have a go at this problem. It's okay to find it hard — struggling builds stronger understanding.

Identical twins share 100% of their DNA. Yet one twin develops Type 2 diabetes at age 40 while the other remains healthy. Suggest how gene expression could explain this difference.

Write your best answer with no support. Consider what could change gene activity without altering the DNA sequence itself.

Use these prompts to structure your answer:

  • What are epigenetic changes?
  • Name two types of epigenetic modification.
  • How could different lifestyles (diet, exercise, stress) lead to different patterns of gene expression?

Complete the paragraph:

Although the twins have the same _______ (DNA sequence), their _______ patterns may differ. Environmental factors such as _______ and _______ can cause _______ changes — modifications that alter gene expression without changing the DNA sequence. For example, _______ groups may be added to DNA, which _______ transcription of certain genes. This means one twin may express genes for insulin resistance while the other does not.

Transcription Factors

Not all genes are active in every cell. Transcription factorsProteins that bind to specific DNA sequences near a gene to control whether RNA polymerase can transcribe it. are proteins that regulate which genes are switched on or off by controlling whether RNA polymerase can bind and begin transcription.

How Transcription Is Initiated

Every gene has a promoterA region of DNA upstream of a gene where RNA polymerase and transcription factors bind to initiate transcription. region — a specific DNA sequence just before (upstream of) the coding region. RNA polymerase cannot bind to the promoter alone; it needs transcription factors to assemble first.

Some genes also have enhancerRegulatory DNA sequences, often far from the gene, that increase the rate of transcription when activator proteins bind to them. sequences. These can be thousands of base pairs away from the gene, but when activator proteins bind to them, the DNA loops so the enhancer comes close to the promoter, boosting transcription.

Interactive — Click each part to learn more
DNA Enhancer Promoter Gene (coding region) Terminator Transcription Factors RNA Pol Activator binds DNA loops Direction of transcription
Exam Tip

Questions often ask you to explain how transcription factors initiate transcription. Key steps: (1) TFs bind to the promoter region, (2) this forms a transcription initiation complex, (3) RNA polymerase can then bind, (4) transcription of the gene begins.

Key Distinction

Activators increase transcription by binding to enhancers and recruiting RNA polymerase. Repressors decrease transcription by blocking TF binding or preventing the initiation complex from forming. This is how different cell types express different genes.

Post-Transcriptional Control

Once pre-mRNA is transcribed, it still needs processing before it can be translated. This is another level where gene expression can be regulated.

Alternative RNA Splicing

Pre-mRNA contains both exonsCoding sequences of a gene that are expressed in the final mRNA. Exons are joined together after splicing. (coding regions) and intronsNon-coding “intervening” sequences within a gene that are removed (spliced out) during RNA processing. (non-coding regions). During RNA processing, introns are removed by a complex called the spliceosomeA large molecular machine made of snRNPs (small nuclear ribonucleoproteins) that catalyses the removal of introns from pre-mRNA., and the remaining exons are joined together.

The key insight: different combinations of exons can be joined together from the same pre-mRNA. This means one gene can produce multiple different proteins. This is called alternative RNA splicing.

Alternative RNA Splicing — Click to toggle
Pre-mRNA Exon 1 Intron 1 Exon 2 Intron 2 Exon 3 Intron 3 Exon 4 Splicing Pathway A Exon 1 Exon 2 Exon 3 Exon 4 → Protein A Splicing Pathway B Exon 1 Exon 3 Exon 4 → Protein B Exon 2 is skipped in Pathway B — different exon combinations produce different proteins from one gene
Real-World Example
The Drosophila Dscam gene can produce over 38,000 different mRNA variants through alternative splicing. This is crucial for the immune system — each neuron can have a unique identity.
This shows that the number of proteins an organism makes far exceeds the number of genes it has. Humans have ~20,000 genes but produce over 100,000 different proteins, largely thanks to alternative splicing. This is a key point for understanding how complexity arises.
Exam Tip

Remember: exons are expressed (they stay). Introns are intervening (they are removed). The spliceosome is the molecular machine that does this work.

Epigenetics

EpigeneticsHeritable changes in gene expression that do not involve changes to the DNA base sequence. Caused by chemical modifications to DNA or histone proteins. literally means “above genetics”. These are modifications that change how genes are expressed without altering the DNA base sequence. They can be inherited through cell division and sometimes between generations.

DNA Methylation

A methyl group (-CH3) is added to cytosineOne of the four DNA bases. When methylated (5-methylcytosine), the gene associated with that region is typically silenced. bases in DNA, usually where cytosine is followed by guanine (CpG sites). This is catalysed by DNA methyltransferase enzymes.

Effect: Methylation typically silences gene expression because:

  • It prevents transcription factors from binding to the promoter
  • It recruits proteins that condense the chromatin, making the DNA less accessible

Methylation patterns can be passed on during DNA replication — this is how differentiated cells “remember” which genes to express.

Histone Modification

DNA is wound around histoneProtein “spools” that DNA wraps around to form nucleosomes. Chemical modifications to histones alter how tightly DNA is packaged. proteins to form nucleosomes. The “tails” of histones can be chemically modified, changing how tightly DNA is packaged.

ModificationEffect on ChromatinEffect on Gene Expression
AcetylationLoosens chromatin (euchromatin)Increases transcription — DNA is accessible to RNA polymerase
DeacetylationTightens chromatin (heterochromatin)Decreases transcription — DNA is tightly wound and inaccessible
MethylationCan loosen or tighten depending on positionCan activate or repress — depends on which amino acid is methylated
Chromatin States — Click to explore
Euchromatin (open) Acetyl groups on histones Gene ON ✓ Heterochromatin (closed) Methyl groups / deacetylated Gene OFF ✗
Making Connections

Epigenetic modifications explain how differentiated cells maintain their identity. A liver cell keeps liver-specific genes active and other genes methylated/silenced. These patterns are copied when the cell divides, so daughter cells remain liver cells.

Flashcard Review

Transcription Factor
A protein that binds to a specific DNA sequence (promoter/enhancer) to regulate the transcription of a gene.
Click to flip
Promoter
Region of DNA upstream of a gene where RNA polymerase and transcription factors bind to initiate transcription.
Click to flip
Enhancer
A regulatory DNA sequence (can be far from the gene) that increases transcription rate when activator proteins bind to it.
Click to flip
Spliceosome
A molecular machine made of snRNPs that removes introns from pre-mRNA and joins exons together.
Click to flip
DNA Methylation
Addition of -CH3 to cytosine bases at CpG sites, typically silencing gene expression by blocking TF binding.
Click to flip
Histone Acetylation
Addition of acetyl groups to histone tails. Loosens chromatin (euchromatin), making DNA accessible for transcription.
Click to flip
Epigenetics
Heritable changes in gene expression without changes to the DNA base sequence, e.g. methylation, histone modification.
Click to flip
Barr Body
An inactivated, condensed X chromosome in female mammals, visible as a dense body at the edge of the nucleus.
Click to flip

Non-coding RNA & X-Inactivation

Not all RNA codes for proteins. Non-coding RNA (ncRNA)RNA molecules that are not translated into protein but have regulatory roles in gene expression. Includes miRNA, siRNA, and lncRNA like Xist. plays important regulatory roles in controlling gene expression.

X-Inactivation

Female mammals have two X chromosomes (XX) while males have one (XY). To prevent a “double dose” of X-linked gene products, one X chromosome in each female cell is inactivated early in embryonic development.

The process is controlled by a long non-coding RNA called XistX-inactive specific transcript — a long non-coding RNA produced from the X chromosome that coats the chromosome from which it is transcribed, triggering its inactivation. (X-inactive specific transcript):

  1. Xist RNA is transcribed from one X chromosome
  2. It coats the chromosome from which it was produced
  3. This recruits proteins that add repressive epigenetic marks (methylation, histone deacetylation)
  4. The coated X chromosome condenses into a Barr bodyThe inactivated, condensed X chromosome visible as a dark-staining body at the edge of the nucleus in female cells.
  5. Most genes on that chromosome are permanently silenced

The choice of which X to inactivate is random in each cell. This means female mammals are mosaics — some cells express the maternal X and some express the paternal X.

Real-World Example
Tortoiseshell cats are almost always female. The patchy coat pattern of orange and black fur is a visible result of X-inactivation — cells with one active X show orange, cells with the other active X show black.
Males (XY) only have one X chromosome, so there is no X-inactivation and no mosaicism. They express whichever colour allele is on their single X chromosome. A male can only be tortoiseshell if he has an XXY genotype (Klinefelter syndrome), which is very rare.

Chromosome Puffs & Cell Differentiation

In the giant polytene chromosomes of Drosophila and Chironomus larvae, regions of active transcription can be seen as “chromosome puffsRegions of decondensed, actively transcribing DNA visible in polytene chromosomes. Different puffs appear in different cell types, showing differential gene expression.” — areas where the chromatin is decondensed and genes are being actively transcribed.

Different cell types show different puffing patterns, providing direct visual evidence that different genes are expressed in different cell types. This is controlled by epigenetic mechanisms.

Visible Thinking: Claim — Support — Question

Consider this statement: “Your lifestyle choices can change how your genes are expressed, and these changes could be passed to your children.”

Make a Claim
Support It
Raise a Question

Fill in the Blanks

1. Transcription factors bind to the region of a gene.
2. sequences can be far from the gene but increase transcription rate.
3. The spliceosome removes from pre-mRNA.
4. Different exon combinations from one gene produce different proteins — this is called alternative RNA .
5. Addition of -CH3 groups to cytosine bases is called DNA .
6. Histone loosens chromatin and promotes transcription.
7. The inactivated X chromosome forms a dense structure called a .
8. The non-coding RNA responsible for X-inactivation is called .
Word Bank

promoter • enhancer • introns • splicing • methylation • acetylation • Barr body • Xist

Sequencing: Gene Expression Control

Drag the steps into the correct order to show how a gene is expressed and regulated.

? Spliceosome removes introns and joins exons
? Transcription factors bind to the promoter
? Polypeptide folds into a functional protein
? Pre-mRNA is transcribed from the DNA template
? Mature mRNA is translated at the ribosome
? RNA polymerase binds and begins transcription

Tiered Practice

Explain how epigenetic mechanisms allow cells with identical DNA to become specialised for different functions. Include reference to DNA methylation, histone modification, and the role of transcription factors. [6 marks]

Use the prompts below to explain how cells become specialised:

  • What role do transcription factors play in gene expression?
  • How does DNA methylation silence genes?
  • How does histone acetylation activate genes?
  • How are these patterns maintained when cells divide?

Complete this model answer:

All cells contain the same _______ but not all genes are expressed in every cell. Transcription factors are proteins that bind to the _______ region of a gene and allow _______ to begin transcription.

Genes that are not needed in a particular cell type can be silenced by DNA _______, where -CH3 groups are added to _______ bases. This prevents transcription factors from _______.

Genes that ARE needed can be activated by histone _______, which _______ the chromatin so RNA polymerase can access the DNA. These epigenetic patterns are _______ when cells divide, so daughter cells maintain the same specialisation.

Exam Questions

2-mark question

State two ways in which epigenetic changes differ from mutations.

1. Epigenetic changes do not alter the DNA base sequence / mutations change the base sequence. (1)

2. Epigenetic changes are often reversible / mutations are permanent changes to the DNA. (1)

4-mark question

Explain how alternative RNA splicing allows one gene to code for more than one protein.

1. A gene contains both exons (coding sequences) and introns (non-coding sequences). (1)

2. Pre-mRNA is transcribed and contains both exons and introns. (1)

3. The spliceosome removes introns, but different combinations of exons can be retained / joined together. (1)

4. Different mature mRNA molecules are produced, which are translated into different polypeptides / proteins. (1)

6-mark question

Discuss the role of epigenetic modifications in controlling gene expression and their significance in cell differentiation. Include reference to DNA methylation, histone modification, and non-coding RNA.

DNA methylation: Methyl groups (-CH3) are added to cytosine bases at CpG sites. This prevents transcription factors from binding to the promoter region, silencing the gene. Methylation patterns are inherited during cell division, maintaining cell identity. (2)

Histone modification: Acetylation of histone tails loosens chromatin (euchromatin), making DNA accessible to RNA polymerase and transcription factors, activating genes. Deacetylation tightens chromatin (heterochromatin), silencing genes. Different patterns of histone modification in different cell types ensure only appropriate genes are expressed. (2)

Non-coding RNA: ncRNA such as Xist plays a role in gene regulation. Xist coats one X chromosome in female cells, recruiting proteins that add repressive epigenetic marks, condensing it into a Barr body. This is dosage compensation — ensuring females do not produce double the X-linked gene products compared to males. This demonstrates how non-coding RNA coordinates large-scale gene silencing. (2)

6-mark Strategy

For “discuss” questions, cover each named topic with: (1) a clear description of the mechanism, (2) its effect on gene expression, (3) its significance/application. Use correct terminology throughout.

Test Yourself

AO1Q1/5
What is the role of a promoter region?
A It codes for the protein product of a gene
B It is the binding site for transcription factors and RNA polymerase
C It is the site where ribosomes attach during translation
D It codes for enhancer proteins
AO2Q2/5
A pre-mRNA molecule has 5 exons. Through alternative splicing, one mature mRNA contains exons 1, 2, 3, and 5, while another contains exons 1, 3, 4, and 5. What can you conclude?
A The gene has mutated between the two splicing events
B Two different proteins can be produced from the same gene
C Exons 2 and 4 must be introns
D One mRNA is faulty and will not be translated
AO1Q3/5
Which epigenetic modification typically silences gene expression?
A Histone acetylation
B DNA methylation
C Phosphorylation of RNA polymerase
D Alternative RNA splicing
AO2Q4/5
A Barr body is observed in a cell. What can you deduce about this cell?
A It is from a male organism
B It is from a female organism with one inactivated X chromosome
C It has undergone a mutation
D It is a cell that is about to divide
AO3Q5/5
Scientists found that identical twins raised apart showed different patterns of DNA methylation. One twin, who smoked, had reduced methylation of a tumour suppressor gene. Suggest what effect this might have.
A The tumour suppressor gene would be silenced, increasing cancer risk
B The tumour suppressor gene would be more active, decreasing cancer risk
C The DNA sequence of the gene would be permanently altered
D The gene would be removed from the genome

How confident do you feel?

Exit Ticket

Three quick questions before you go:

1. Describe the role of transcription factors in gene expression.

2. Explain how one gene can produce multiple different proteins.

3. Describe how DNA methylation and histone acetylation have opposite effects on gene expression.

Vocabulary Grid

Transcription factor
Protein that binds to promoter/enhancer to control whether a gene is transcribed.
Promoter
DNA region upstream of a gene where TFs and RNA polymerase bind.
Enhancer
Distant regulatory sequence that boosts transcription when activators bind.
Spliceosome
Molecular machine (snRNPs) that removes introns and joins exons.
Alternative RNA splicing
Different exon combinations from one pre-mRNA producing different proteins.
Epigenetics
Heritable changes in gene expression without altering the DNA sequence.
DNA methylation
Addition of -CH3 to cytosine at CpG sites; silences gene expression.
Histone acetylation
Acetyl groups loosen chromatin (euchromatin); promotes transcription.
Barr body
Inactivated, condensed X chromosome in female cells.
Xist
Long non-coding RNA that coats and silences one X chromosome in females.

Self-Assessment Checklist

I can explain how transcription factors regulate gene expression
I can describe the roles of promoters and enhancers
I can explain alternative RNA splicing and its significance
I can describe how DNA methylation silences genes
I can explain the effect of histone acetylation on chromatin structure
I can describe X-inactivation and the role of Xist RNA
I can explain what a Barr body is and why it forms
I can link epigenetics to cell differentiation