1. Topic 12 Genetic Engineering
Selective Breeding vs. Genetic Engineering
Polymerase Chain Reaction, Gel Electrophoresis & DNA Sequencing
HGP & DNA Profiling in Forensics
Stem Cells & Cloning
DNA Probes, PGD & Gene Therapy

2. Selective Breeding vs. Genetic Engineering
Topic 12: Lesson 1
Resources:
Read Chapter A pp
Kahn Academy – Restriction Enzymes
Lab – Restriction Enzymes
Selective Breeding vs. Genetic Engineering

3. Selective Breeding = Artificial Selection
Humans have been using artificial selection for thousands of years

4. Artificial Selection in Pets & Livestock
Selective breeding takes many generations to achieve desired results, but can be achieved without much technology

5. Genetic Engineering
Directly inserting, removing, or changing an organism’s DNA using technology
Genetically Modified Organisms (GMOs) contain DNA from other organisms (transgenic organisms)

6. NOT Examples of GMOs
Grafting
Hybrid animals

7. Example of GMO: Bt Corn
European corn borer lays eggs in corn plants and the larvae feed on the corn leaves
Scientists inserted a gene from a soil bacteria, Bacillus thuringiensis, that is deadly to the corn borer in the corn

8. How to Make a Transgenic Organism
Obtain source DNA (gene)
Obtain a plasmid (circular DNA) from bacteria
Use restriction enzymes to cut out the gene of interest and to cut the plasmid to receive the gene
Mix source DNA gene with plasmid to form recombinant DNA
DNA ligase will seal the phosphodiester bonds
Insert recombinant DNA into host cell

9. Restriction Enzymes

10. Transgenic Plant with DNA from Bacterium

11. Polymerase Chain Reaction, Gel Electrophoresis & DNA Sequencing
Topic 12: Lesson 2
Resources
Read Chapter 11.2B-C pp
Virtual Lab – PCR
DNA Learning Center – PCR
MIT Open Courseware – PCR
Kahn Academy – PCR
Virtual Lab – Gel Electrophoresis
Kahn Academy – Gel Electrophoresis
Kahn Academy – DNA Sequencing
AK Lectures – Sanger DNA Sequencing
Polymerase Chain Reaction, Gel Electrophoresis & DNA Sequencing

12. PCR Amplifies Genes
95°C
54°C
72°C

13. PCR = Copying DNA by Alternating Temperatures
One gene can be copied millions of times within 1-2 hours
A three step process is repeated for times
A thermal cycler machine controls temperature changes
STEPS OF PCR
Denaturation (95°C; 201.2°F) – the double stranded DNA opens into two pieces of single stranded DNA
Annealing (54°C; 129.2°F) – primers pair up with the single stranded DNA template; DNA (Taq) polymerase begins to copy the template
Extension (72°C; 161.6°F) – DNA (Taq) polymerase is at its optimum temperature and completes copying the double stranded DNA

14. Uses of PCR First step in each of the following practices
Establish paternity
DNA fingerprinting
Study human evolution
Diagnose genetic diseases
Clone DNA of dead organisms

15. Gel Electrophoresis

16. Sorting Molecules by Size

17. DNA Sequencing
Finding the Order of Nucleotides in DNA

18. Using Gel Electro for DNA Sequencing

19. Different People Produce Different Bands on Gels
SNP/RFLP
STR

20. Paternity Testing

22. Human Genome Project & DNA Profiling in Forensics
Topic 12: Lesson 3
Resources
Read Chapter 11.2B pp
Bozeman Science – DNA Fingerprinting
Naked Scientists – How Does DNA Fingerprinting Work?
Activity – Who Killed Sharon?
Human Genome Project & DNA Profiling in Forensics

23. Human Genome Project Aim to sequence the 3.2 billion base pairs
Completed in 2003
Found
25,000 genes
400,000 proteins
1.5% of DNA codes for protein

24. Finding Variation in DNA
Each human’s DNA is 99.9% identical
Large variation is found in between the genes in non-coding regions

25. FBI Using DNA Fingerprinting
The FBI uses 13 different STRs to determine the matching identity of a person

26. Stem Cells & Cloning Topic 12: Lesson 4 Resources
Read Chapter 11.3A-B pp
Worksheet – Stem Cells & Cloning
NOVA – Stem Cell Video
NOVA - Stem Cell Poll
Module 6 – Contrasting Cases of Animal Modifications
Stem Cells & Cloning

27. From Zygote to Morula to Blastula to Gastrula

28. 3 Germ Layers

29. What is a Stem Cell?

30. Stem Cells Embryonic Stem Cells Adult Stem Cells
Totipotent – give rise to all cell types in the body
Derived from zygote cell or morula (first 8 cells)
Pluripotent – give rise to many but not all types of cells in the body
Derived from Inner Cell Mass of blastocyst

33. Cloning As a Means of Asexual Reproduction
Many organisms using cloning to create identical copies of themselves
Plants, fungi, bacteria, and some animals use asexual reproduction
Human can use cuttings to clone fruit trees and flowers

34. Gene Cloning
DNA/Molecular/Recombinant/Gene – DNA from one organism is combined with a vector and then put into a host cell
mRNA from eukaryotic cell is copied into cDNA
cDNA will be spliced into a vector plasmid with a restriction enzyme
The recombinant vector (cDNA + vector) is inserted into a host cell
Once in the host cell, the recombinant vector will replicate before cell division
The cell will make the proteins encoded in the cDNA

35. Reproductive Cloning (SCNT)

36. Therapeutic Cloning
Stem cells are extracted from a 5 day old embryo; the embryo is destroyed but the stem cells can transform into any of the 220 human cell types

37. DNA Probes, PGD & Gene Therapy
Topic 12: Lesson 5
Read Chapter 11.4 pp
PGD Video – Genetics & IVF Institute
Do you want to know your chances of getting Alzheimer's disease?
Module 4 – Max’s Case
DNA Probes, PGD & Gene Therapy

38. Probes Look for Genes
DNA probes detect specific nucleotide sequences in DNA
DNA probes are ssDNA strands that are complementary to a certain gene
Labeled with a fluorescent tag of radioactive isotope
Target sequence DNA is heated so it separates
DNA probe binds to region that it is complementary to
Scientists detect radioactivity or use a fluorescent light to determine if the gene is present

39. FISH

40. Preimplantation Genetic Diagnosis
If two carriers of a particular disease want to be sure that they have a baby that is disease free, PGD can reduce the odds
in vitro fertilization occurs (IVF)
Morula is formed and one of the cells is selected for diagnosis
PCR amplifies the region of DNA where the suspected allele may be
DNA probe determines if the alleles are present of the recessive condition
Embryo can be discarded or implanted back into the woman’s body if it is considered a healthy cell

41. Genotyping Using Gel Electro

42. Testing 1 of 8
What bioethical issues present themselves when PGD is applied?
Who should do PGD?
Mothers over 35
Repeated miscarriages
Previous pregnancy with chromosomal abnormality
X-linked carrier mothers
Unexplained infertility
IVF failures

43. Fixing Bad Genes
Most genetic diseases do not have a cure, but gene therapy could provide new treatment options
Gene therapy corrects defective genes
Swaps for bad copy, reverses mutation, turns off the gene
A vector (vehicle) that can deliver a gene to all of the cells that are “infected”
Viruses and plasmids are the two most commonly used vectors

44. Treating Cystic Fibrosis with the Cold Virus
Viruses can act as a vector for CF patients
The virus is introduced to the lung cells with a normally functioning gene
The CF gene is replaced by the normal gene

45. RNA Interference
RNAi can prevent translation of mRNA silencing the gene
RNAi bonds to a complementary mRNA strand to make a double stranded and nonfunctional mRNA