DNA: found in the nucleus, in the form of chromosomes

Locus : gene’s position on a chromosome

Allele: different variation of genes

DNA is made up of… nucleotides

Nucleotides:
- 5 carbon sugar (deoxyribose)
- 1 phosphate group
- 1 nitrogenous base
Nitrogenous base:
- Adenine, Thymine
- Guanine, Cytosine
Base Pairing:
Hydrogen bonds only form between Adenine and Thymine and Guanine and Cytosine

DNA Replication
Helicase: unzips DNA by breaking the bonds of the base pairs
DNA Polymerase: principal DNA involved in DNA replication, joins nucleotides together
Winding Enzymes: zips 2 strands together

RNA: uses genetic code from DNA to create protein, single stranded, contains Uracil instead
of Thymine
- mRNA (messenger RNA): carries copies of instructions for assembling amino acids to
proteins, from DNA to cells
- rRNA (ribosomal RNA): makes up ribosome, links amino acids together
- tRNA (transfer RNA): moves amino acid to ribosome during building of protein

Introns: sequences of DNA that is not involved in coding for proteins

Exons: sequences of DNA that is involved in coding for proteins
Translation: another word for protein synthesis
Genome: the entirety of human’s genetic info
DNA Sequencing: technique by which the precise order of nucleotides can be determined
Why sequence DNA?
1. Determine protein produced
2. Determine family pedigree
3. Identify genetic faults
4. Compare taxonomy
5. Forensic
6. Medical diagnosis

Human genome project: international effort to sequence the entire human genome
- Gene identification: how many and what genes, understand functions of genes
- Phylogenetic: identify similarities and differences between the genome of other
organisms, understands evolutionary relationships
- Medical Diagnosis: identify changes in sequence, drug targeting/ gene therapy

Pyrosequencing: detects activity of DNA polymerase. Sequences 1 single strand of DNA by

synthesizing the complementary strand one base pair at a time.
Proteome: the entirety of proteins that a cell is able to make
Proteomics: identifies all the proteins in a cell, understands its functions and interactions,
useful in medical research
Simple organisms: easy to determine proteome bcs it does not contain introns
Complex organisms: knowledge of gene can’t easily be translated to proteome
because…
o Hard to distinguish between Extron and intron
o Makes it hard to find code sections

Genetic Engineering: make changes in the DNA of a living organism

Recombinant DNA: (fragments) DNA from one organism that is attached to another
Transgenic DNA: recipient of the recombinant DNA
Turning DNA into fragments…
1. Reverse Transcription: because finding the DNA would be too hard, we just find the
RNA; cell that is expressing the gene would have an RNA with complementary
sequence, using the enzyme reverse transcriptase, we can make DNA from RNA
2. Restriction: Hard bcs we need to actually find the gene.
 Step 1: break open cell and isolate DNA
 Step 2: incubate DNA with restriction endonuclease enzyme, that will
release the cut out of the gene which cuts at specific sites
 Restriction endonuclease enzyme also recognizes specific palindromic
sequences in the DNA they cut
 This sequence is palindromic, meaning that the same cut will be made
on both strands
Sticky Ends: enable DNA from one organism to be combined with DNA from another
organism making a recombinant DNA.

3. Gene Machine
 We are able to synthesize any fragments of the DNA using the gene machine,
without the need of a template gene.
 You are able to design the DNA strand

Transformed Cells: genetic alteration of a cell resulting from incorporation of external

genetic material. Cell takes DNA from outside the cell and becomes a component of the
cell’s DNA.

Transformed microrganisms: cut desired gene from donor with restriction endonuclease,
cut plasmid vector, insid plasmid into bacteria, identify transformed bacteria
Transformed Animals: cut DNA fragment, make DNA fragment using reverse transcriptae,
insert into early embryo or eggs
Transport Plants: cut DNA fragment, Insert into plasmid and transfer into bacteria, Bacteria
“infects” plant transferring plasmid with desired gene
 The Central Dogma of Molecular Biology states that DNA makes RNA makes proteins

 Replication: copying DNA

 Transcription: DNA copied (transcribed) to mRNA

 Translation: mRNA directed protein synthesis

 Promoter region: a region of DNA that leads to initiation of transcription of a
particular gene

Defective Genes:

- Genetic disorders arise from mutated alleles

- Dominated disorders need only 1 allele to cause the disease

- Recessive disorders need both alleles to be defective for the disease to arise

Gene Therapy:

- Using recombinant gene technology to treat disease

- Adding the gene/ gene augmentation: introduce the functioning gene, the correct
gene product masks the effect of the non-functional protein

- Inhibiting the gene: introduce a piece of DNA which inserts itself in the middle of the
defective gene, gene can no longer be transcribed, malfunctioning protein is no
longer produced

- Successes: parkinsons, haemophilia, cystic fibrosis, SCID, Thalassaemia, LCA

How do we deliver genes?

- By cloning them and transporting them through a vector (vehicle) that takes the
genes into the cells that needs the genes.

- Delivery methods:

 In vivo: directly into body, virus is modified to prevent it multiplying and

attacking cells, mixed with a liquid and used as an inhaler, virus inserts DNA
into target cells

 Ex vivo: remove cells, add gene, replace cells

  Retrovirus: inserts gene into host DNA passed on

 Adenovirus: DNA left in free nucleus- not replicated with division

 Liposome: non-specific

 Naked DNA: non-specific injection

Multicellular Organisms: organisms made of many different types of cells that are
specialized to their function

- Grows and develops by 2 basic processes:

 Cell division

 Differentiation: cells become specialized

Cells:

- All contain a copy of an organism’s DNA

- All cells contain the same genes

- Every cell is capable of making every product that the body can produce

Differentiation:

- Although cells contain all genes, only a small part of it is expressed

- Different specialized cells have different combinations of genes that can be switched
on or off

 Meaning that only certain genes are expressed in one cell at any time

- Differentiation is permanent, a one way process

- The type of specialized cell produced is determined by the genes that are switched
on in that cell

- Once specialized, a cell cannot go back and become a stem cell

Specialized cells: produce different proteins

Proteins modify the cell:

- Determine the structure

- Control the processes

Specialisation: changes to the cell as a result of the proteins produced

All specialized cells originate from stem cells

stem cells: unspecialized cells that can develop into other types of cells

potency: the ability to differentiate into specialized cells

self-renewal: the ability to replicate indefinitely

more genes are switched on and fewer switched off in differentiation

2 features of stem cells:

- Replace themselves/keep dividing/replicate

- Can differentiate/ develop into other cells

Totipotent: can differentiate into any embryonic stem cell

- Capable of giving rise to whole organism (includes body cell or support cells like
placenta)

- Only present in very early mammalian embryos

- E.g: zygote

Pluripotent: can differentiate into any kind of body cell

- Can’t differentiate into cells of placenta or umbilical cord uncapable of making

whole organism

- E.g: embryonic stem cells

Adult stem cells:

- Obtained from the body tissue of an adult

- Obtained by simple operation: low risk but uncomfortable

Multipotent: give rise to a limited number of other cell types

- Are adult stem cells

- E.g: haemotopoietic bone marrow give rise to red blood cells, white blood cells and
platelets

Unipotent: least potent

- Can differentiate into only one type of cell

 - Generally provide replacement in different body parts

- E.g: heart stem cell

Cardiomyocytes: heart muscle cells

- Specialized cells and unable to divide and replicate in mature mammals

- Problem: if heart becomes damaged, these cells can’t be repaired

- Scientists now believe that cardiomyocytes can be replaced by new cardiomyocytes

derived from a small supply of unipotent cells in the heart

Yamanaka:

Showed that specialized adult cells could be turned into stem cells in mice

Requires reprogramming with 4 transcription factors

Induced Pluripotent stem cells can make any type of cell in the body/ reprogrammed
specialized cells made to express transcription factors normally associated with pluripotent
stem cells

In theory, this shows that any cells of the body can be turned into a pluripotent cell.

Benefit?

- No ethical dilemma

- Stem cells from patients own cell (no rejection)