1. UNIT VII – GENOMICS & CANCER
Big Campbell – Ch 17, 18, 20
Baby Campbell – Ch 10, 11, 12
Hillis – Chp 12, 13,

2. I. EUKARYOTIC GENE EXPRESSION REGULATION, cont Epigenetics
Definition
study of heritable changes in gene expression that do not involve changes to the DNA sequence; change in phenotype without a change in genotype.
Due to: DNA methylation, histone modification and non-coding RNA (ncRNA)-associated gene silencing
Illnesses, behaviors, & other health issues have been linked to epigenetic mechanisms, including cancers of almost all types, cognitive dysfunction, respiratory, cardiovascular, reproductive, autoimmune, and neurobehavioral illnesses.

3. I. EUKARYOTIC GENE EXPRESSION REGULATION, cont Epigenetics

4. I. EUKARYOTIC GENE EXPRESSION REGULATION, cont Epigenetics

5. II. GENOMICS Human Genome Project Goals
Identify all the approximately 20,000-25,000 genes in human DNA
Determine the sequences of 3 billion chemical base pairs that make up human DNA
Store information in databases
Improve tools for data analysis
Transfer related technologies to the private sector
Address the ethical, legal, and social issues (ELSI) that may arise from the project.
Completed early and under-budget in 2003

6. II. GENOMICS, cont
Chromosome 6, p Arm

7. II. GENOMICS, cont Transposons
DNA sequences that move from one location in the genome to another
“Jumping Genes”

8. II. GENOMICS, cont Human Genome Applications
SNPs – single nucleotide polymorphisms
Single base-pair that shows variation in a significant % of population
SNPs that alter the fragment length following exposure to restriction enzymes called RFLPs (restriction fragment length polymorphisms)
Genetic markers

9. II. GENOMICS, cont Human Genome Applications, cont
STRs – short tandem repeats
Short segments of DNA that are highly repetitive, polymorphic
Repeat patterns are inherited
Useful for DNA fingerprinting

10. III. BIOTECHNOLOGY, cont Tools, cont
Polymerase Chain Reaction (PCR)
In vitro method of amplifying small amounts of DNA
DNA is heated to separate the double helix.
Mixture is allowed to cool, DNA primers attach to target
Heat-stable polymerase is used to extend the primers in the 5’–3’ direction.

11. III. BIOTECHNOLOGY, cont Tools, cont
Southern Blotting
Designed by Dr. Southern
Detects particular DNA sequences
Northern Blotting
Detects particular mRNA sequences
Western Blotting
Used to detect proteins

12. III. BIOTECHNOLOGY, cont Tools, cont
cDNA - complementary DNA
Procedure for “cloning DNA” that uses mRNA, reverse transcriptase

13. III. BIOTECHNOLOGY, cont
Recombinant DNA
DNA containing nucleotides from other sources
Process utilizes restriction enzymes that make jagged cuts in DNA; creates sticky ends
When DNA from different sources treated with same restriction enzyme, sticky ends “mix & match”
Often use reporter genes to determine success; for example, ampicillin resistance

14. III. BIOTECHNOLOGY, cont
DNA Microarray Assays
AKA DNA Chips
Test used to determine gene function, gene interactions
May be used to determine agressiveness of cancers, method of treatment, etc

15. III. BIOTECHNOLOGY, cont
Gene Cloning
Typically plasmid vector
Plasmid isolated from bacterial cell
Foreign DNA inserted into plasmid
Plasmid returned to bacterial cell; described as recombinant bacterium
Foreign gene is cloned as bacteria reproduce
Common bacterium used for plants is Agrobacterium tumefactiens

16. III. BIOTECHNOLOGY, cont
A CLOSER LOOK AT GENE
CLONING

17. III. BIOTECHNOLOGY, cont
Therapeutic Cloning ES
iPS

18. III. BIOTECHNOLOGY, cont
Reproductive Cloning
Nuclear Transplantation
Process of using unfertilized egg cell & replacing nucleus with DNA
In 1997, scientists were able to produce first reproductive clone, “Dolly”, by culturing somatic cells in a nutrient-poor medium to de-differentiate them and force them back to totipotency.
Reproductive cloning in animals has enjoyed limited success.

19. III. BIOTECHNOLOGY, cont
Transgenic Organisms

20. III. BIOTECHNOLOGY, cont
RNAi
Based on principal of microRNA
Exogenous small-interfering RNA (siRNA) synthesized taken up by cells or gains entrance via a viral vector
Complementary to mRNA target
Translation is blocked
Has been used to block production of growth factors in certain cancers