Key Points: Human Biology ATAR - Recombinant DNA Technology, Gene Therapy, Cell
Replacement Therapy
🧬 Recombinant DNA Technology
Involves inserting a gene of interest (e.g., human insulin or human growth hormone gene)
into a bacterial plasmid.
Key steps:
1. Identify and isolate the gene (e.g., human insulin gene) on chromosome
2. Use restriction enzyme to cut the gene at specific sites, creating sticky ends.
3. Use same restriction enzyme to cut out a section of a plasmid (also creating sticky
ends in the plasmid)
4. Ligate / insert the gene into the plasmid using DNA ligase.
5. Insert recombinant plasmid into a host bacterial cell (transformation).
6. Bacteria (grown in culture) clone the plasmid and express the human gene to
produce / synthesize the protein (e.g., insulin, hGH).
7. Protein is harvested and purified for medical use.
8. Helpful for producing large scale amounts of proteins (e.g., insulin, hGH).
Applications in syllabus:
o Producing hormones (e.g., insulin for diabetes, human growth hormone for
dwarfism).
Previous exam question below
🧬 Gene Therapy
1. Identify and isolate the normal gene:
o For diabetes: isolate a functioning insulin gene.
o For cystic fibrosis: isolate a normal CFTR gene (which codes for the chloride channel
protein).
2. Insert the gene into a vector:
o The gene is inserted into a carrier (vector), often a virus (e.g., adenovirus,
retrovirus) that’s been modified to be harmless.
o The vector carries the gene into target cells.
3. Prepare target cells:
o Identify and isolate the target cells:
For diabetes: cells in the pancreas (beta cells) or sometimes liver cells.
For cystic fibrosis: lung epithelial cells.
4. Deliver the gene to target cells:
o Introduce the vector containing the gene into the body:
In vivo (directly into the patient) → e.g., inhaling a spray of viral particles
into lungs for CF.
Ex vivo (cells modified outside body and then transplanted) → less common
for CF and diabetes.
o The vector infects the target cells, inserting the normal gene into their DNA.
5. Expression of the gene:
o Target cells begin producing the correct protein:
Diabetes: cells make insulin.
Cystic fibrosis: cells make functional chloride channel proteins.
6. Monitor effectiveness:
o Doctors monitor protein production, symptom improvement, and potential side
effects (e.g., immune response to vector).
7. Repeat treatment if necessary:
o Because some gene therapies aren’t permanent, treatment may need to be
repeated as cells divide or die.
✨ Key differences between diabetes and cystic fibrosis applications:
Diabetes: goal is to restore insulin production to lower blood glucose levels.
Cystic fibrosis: goal is to correct defective chloride transport to reduce thick mucus buildup.
Applications in syllabus:
Gene therapy can treat diseases like diabetes mellitus.
🧬 Cell Replacement Therapy
Steps in Cell Replacement Therapy
1️. Source of Stem Cells Identified
Obtain pluripotent stem cells (e.g. embryonic stem cells or induced pluripotent stem cells
[iPSCs]).
These cells have the ability to differentiate into various types of neurons.
2️. Stem Cells Cultured and Grown
Stem cells are cultured in controlled lab conditions.
They’re encouraged to multiply to produce sufficient numbers of cells for therapy.
3️. Directed Differentiation
Stem cells are chemically or genetically induced to develop into:
o Dopamine-producing neurons (for Parkinson’s disease).
o Cholinergic neurons or other neuron types (for Alzheimer’s disease).
Specific growth factors and signalling molecules guide this process.
4️. Testing and Quality Control
Cells are tested for:
o Purity (no undifferentiated cells that could form tumours).
o Functionality (ability to produce neurotransmitters like dopamine or acetylcholine).
o Genetic stability.
5️. Preparation for Transplantation
Cells are prepared in a sterile, transplant-ready form (may be in suspension or on scaffolds).
Immunosuppressive protocols may be planned to prevent rejection (if cells are not
autologous).
6️. Transplantation into the Patient’s Brain
Neurosurgeons inject the cells into target brain regions:
o Substantia nigra (for Parkinson’s—replaces lost dopamine neurons).
o Hippocampus or cortex (for Alzheimer’s—attempting to replace lost neurons
involved in memory and cognition).
Stereotactic surgery (precise targeting guided by imaging) is used.
7️. Post-Transplant Monitoring and Support
Monitor patient for:
o Cell survival.
o Integration into neural circuits.
o Functional improvement (motor control in Parkinson’s, memory/cognition in
Alzheimer’s).
Monitor for adverse effects (e.g. immune rejection, tumour formation).
✏️Differences in Focus:
Disease Target Cells Target Brain Region
Parkinson’s Disease Dopamine-producing neurons Substantia nigra
Alzheimer’s Disease Cholinergic neurons / cortical neurons Hippocampus / cerebral cortex
Current challenges:
Ethical issues (especially embryonic stem cells).
Risk of tumour (teratoma) formation.
Immune rejection (esp. non-autologous cells).
Ensuring transplanted neurons integrate and function properly.
Summary
Aims to replace damaged or lost cells with healthy, functioning ones (using stem cells or
engineered cells).
Applications:
o Diabetes: replacing destroyed pancreatic beta cells to restore insulin production.
o Alzheimer’s disease: replacing neurons lost due to neurodegeneration.
o Parkinson’s disease: replacing dopamine-producing neurons in the substantia nigra.
Current limitations: sourcing cells, immune rejection, risk of tumour formation.
Exam example:
Question: "Describe how cell replacement therapy is being used to treat Parkinson’s
disease."
Answer: Transplanting dopamine-producing neurons or precursors into the brain to restore
dopamine levels and improve motor function.
📘 Exam Questions & Marking Key Summaries:
Topic Example Exam Question Key Points in Answer
Describe process to produce hGH Identify gene, cut with restriction enzyme,
Recombinant
using recombinant DNA plasmid cut same enzyme, ligate, transform
DNA
technology bacteria, clone, express
What type of therapy used to
Gene Therapy inject DNA into rats to correct Gene therapy, replaces faulty gene
gene?
Cell Describe how cell replacement
Transplant dopamine-producing neurons to
Replacement therapy is used to treat
replace those lost in substantia nigra
(PD) Parkinson’s
How could gene therapy and cell
Combined Gene Insert healthy gene into replacement cells
replacement therapy be
+ Cell before transplanting to treat disease
combined?
✏️Syllabus Links
The syllabus explicitly includes these under Science as a Human Endeavour:
Gene therapy can treat diseases like diabetes mellitus.
Recombinant DNA technology produces hormones and vaccines.
Cell replacement therapy potential for treating Alzheimer’s and Parkinson’s disease.
