BiotechnologyBiotechnology

Genetic EngineeringGenetic Engineering

The use of microorganisms or biological substances to perform specific industrial or manufacturing processes

Recombinant DNARecombinant DNAGenes from different species are Genes from different species are combined and placed into host cellscombined and placed into host cells

• Example:Example:• Green Fluorescent Protein (GFP)Green Fluorescent Protein (GFP)

• Found in jellyfish, firefliesFound in jellyfish, fireflies

Making Recombinant DNAMaking Recombinant DNA

2. Insert gene of 2. Insert gene of interest into a interest into a plasmidplasmid•(GFP) (GFP) gene + vectorgene + vector= = recombinant plasmidrecombinant plasmid•Vector is a small, Vector is a small, circular piece of DNA circular piece of DNA •Plasmid transports Plasmid transports

target (GFP) gene target (GFP) gene to to host cellhost cell

3. Transfer plasmid into 3. Transfer plasmid into cells of host cells of host organism organism

GFPGFP Plasmid gets inserted into Plasmid gets inserted into EMBRYONICEMBRYONIC catcat cells cells

Cat’s Chromosomes within nucleus

Plasmid

44. Transformation of host . Transformation of host organism’s cellsorganism’s cells•Plasmid will eventually be Plasmid will eventually be incorporatedincorporated into host into host organism’s DNA organism’s DNA •In our example, cat will now In our example, cat will now be able to produce GFPbe able to produce GFP

• AKA the cat will nowAKA the cat will now GLOW!!!GLOW!!!

Mr. Green Genes,Mr. Green Genes,a transgenic cata transgenic cat

Green fluorescence Green fluorescence protein, or GFP, was protein, or GFP, was inserted into his genome inserted into his genome using recombinant DNA using recombinant DNA technology.technology.

GFP fluoresces (glows) GFP fluoresces (glows) when exposed to a when exposed to a blacklightblacklight

Other uses for recombinant Other uses for recombinant DNA:DNA:

1. Agriculture:-Crops: Disease protection, drought & frost

resistance, increased growth-Livestock: Increased muscle mass, hormone

acceleration, drugs in milk2. Pharmaceuticals: HGH, Insulin: insertion of human gene into bacteria for cheap proteins.

Instead of putting plasmid into eukaryotic cells, you can put it in bacteria, which will undergo transformation, and produce a bunch of the protein for human use

INSULIN PRODUCTION!

Other applications of Other applications of biotechnology…biotechnology…

Genetically modified foodGenetically modified foodThese fish are the same age, but the one on the bottom is growing much slower. The fish on the top has been genetically modified to grow faster.

These tomatoes were grown in the same field. The tomatoes on the left have been eaten by pests while the tomatoes on the left are untouched because they were genetically modified to be pest resistant.

ProsProsGenetically Modified (GM) crops can be made resistant to viruses, fungi and bacterial growth.

-grow faster.

-pest-resistant

-tolerate extreme weather conditions

-engineered with added vitamins and minerals

ConsCons

What are the long term health What are the long term health effects?effects?Can GM foods cause new Can GM foods cause new allergies?allergies?What are the effects on the What are the effects on the environment?environment?What’s next…genetically modified What’s next…genetically modified animals?animals?

How can you tell your food is How can you tell your food is genetically modified?genetically modified?

A warning label is currently not requiredA warning label is currently not required

To know, look for common GM ingredientsTo know, look for common GM ingredients

Soy lecithinSoy lecithin

FructoseFructose

DextroseDextrose

Grapeseed oilGrapeseed oil

Cottonseed oilCottonseed oil

Most corn syrups or corn starchesMost corn syrups or corn starches

Some foods are labeled GMO FreeSome foods are labeled GMO Free

More common than you think….

DNA can be used in Crime DNA can be used in Crime Scene InvestigationScene Investigation

DNA FingerprintingDNA Fingerprinting• Each person has their own unique Each person has their own unique

DNA “fingerprint” DNA “fingerprint” except for identical except for identical twinstwins

• Used for forensics, paternity testing, Used for forensics, paternity testing, and identification of remainsand identification of remains

• Small quantities of Small quantities of blood, semen, or blood, semen, or body tissue body tissue can be tested for DNA can be tested for DNA base sequencesbase sequences

DNA Fingerprints are made DNA Fingerprints are made by Gel Electrophoresis by Gel Electrophoresis

Gel Electrophoresis Gel Electrophoresis is a technique used is a technique used to separate to separate molecules based on molecules based on sizesize

The size of the The size of the fragments are fragments are unique to each unique to each person so it creates person so it creates a DNA fingerprinta DNA fingerprint

How does gel electrophoresis How does gel electrophoresis work?work?1.1. Restriction enzymes Restriction enzymes cut DNAcut DNA

2.2. Pieces of DNA are run Pieces of DNA are run through a gelthrough a gel

3.3. The pieces of DNA are The pieces of DNA are separated by separated by weightweight and and make a banding pattern that is make a banding pattern that is unique to an individual unique to an individual (fingerprint)(fingerprint)

4.4. Bands that align with each Bands that align with each other indicate a matchother indicate a match

Gel ElectrophoresisGel Electrophoresis

A case for MauryA case for Maury

• Shown at the right are Shown at the right are DNA "fingerprints" DNA "fingerprints" from a new mother from a new mother (M), her recently born (M), her recently born child (C), and two child (C), and two possible fathers (F1, possible fathers (F1, Ross and F2, Rick).

• What do you think - What do you think - is is Ross (F1) or Rick (F2) the father of the child?

M C F1 F2

A crime scene caseA crime scene case• Shown to the right are Shown to the right are

DNA "fingerprints" DNA "fingerprints" taken from seven (7) taken from seven (7) suspects (the numbered suspects (the numbered columns). columns).

• The bloodstain in the The bloodstain in the middle is from the middle is from the crime scene, ostensibly crime scene, ostensibly from the killer. Which from the killer. Which suspect is guilty?suspect is guilty?

How many genes do How many genes do humans have? What do humans have? What do

they code for?they code for?

Human Genome ProjectHuman Genome Project1990-20031990-2003

Goals:Goals:

Identify ALL genes in human DNAIdentify ALL genes in human DNA

Determine sequence of the base pairsDetermine sequence of the base pairsWe thought there were BILLIONS of bases in We thought there were BILLIONS of bases in the human genomethe human genome

Store all information in databaseStore all information in databaseUse information for disease/drug research, to Use information for disease/drug research, to compare human genome to other organismscompare human genome to other organisms

Obama, State of the Union 2013: “For every Obama, State of the Union 2013: “For every $1 we spent mapping the human genome, we $1 we spent mapping the human genome, we got $104 back”got $104 back”

With the HGP, we have discovered which genes are affected when you have the above diseases

What did we learn?What did we learn?We only have about 30,000 genesWe only have about 30,000 genes

A huge portion of our genome is non-A huge portion of our genome is non-coding, meaning it does not code for coding, meaning it does not code for proteinsproteins

In order to make huge advances in In order to make huge advances in diseases like cancer, the environment has diseases like cancer, the environment has to be taken into accountto be taken into account

Gave rise to fields such as epigenetics Gave rise to fields such as epigenetics

Is it possible to fix a Is it possible to fix a genetic mutation genetic mutation

using using biotechnology??biotechnology??

…Yes! It’s called gene therapy!

Gene TherapyGene Therapy

• Replacement of defective genes Replacement of defective genes with healthy geneswith healthy genes

• Prevents or treats diseasePrevents or treats disease• Examples: cystic fibrosis, Examples: cystic fibrosis,

hemophilia, sickle-cell anemia, hemophilia, sickle-cell anemia, immune-deficienciesimmune-deficiencies

CloningCloning

Cloning is the production of Cloning is the production of genetically identical organisms from genetically identical organisms from single cellssingle cells

First gene cloned mammal= Dolly First gene cloned mammal= Dolly the sheepthe sheep

Egg cellPin

Needle

How it works…How it works…

Egg cell is enucleated (nucleus removed) and Egg cell is enucleated (nucleus removed) and replaced with a nucleus taken from an adult replaced with a nucleus taken from an adult somatic cellsomatic cell

The egg is then placed in the reproductive The egg is then placed in the reproductive system of a surrogate mother, where it system of a surrogate mother, where it develops normallydevelops normally

Surrogate gives birth to offspring that is Surrogate gives birth to offspring that is identical to nuclear donoridentical to nuclear donor

http://learn.genetics.utah.edu/content/cloning/whatiscloning/

Animals that have been clonedAnimals that have been cloned• FishFish• Cats Cats

• CattleCattle• DeerDeer• DogDog• FerretFerret• Fruit Fruit

FliesFlies• GoatGoat• HorseHorse

•Mice•Mule•Pig•Rabbit•Rat•Rhesus Monkey•Sheep•Water buffalo•Wolf

Pets clonedPets cloned

These dogs, Mira and Missytoo, were both cloned from a dog that died in 2002. They are owned by Lou Hawthorne of California. Mr. Hawthorne owns BioArts, a company

with offices in California and laboratories in South Korea. The company offers pet cloning for around $150,000.

Current debateCurrent debateDecember 28December 28th, th, 2007 - FDA said cloned 2007 - FDA said cloned animals are safe to produce and eatanimals are safe to produce and eat

Waiting for consumer approval - has not Waiting for consumer approval - has not happened yethappened yet

United States bans United States bans humanhuman cloning of all types cloning of all types including organs.including organs.

Stem CellsStem CellsIs it possible to grow a leg or an Is it possible to grow a leg or an arm from just one cell?arm from just one cell?

Stem CellsStem CellsStem cells are cells that are Stem cells are cells that are undifferentiatedundifferentiated

They can become any cell, in any part They can become any cell, in any part of the body, and can divide without of the body, and can divide without limit to replenish other cells as long as limit to replenish other cells as long as the person is alivethe person is alive

Found in all multi-cellular organismsFound in all multi-cellular organisms

2 types of stem cells2 types of stem cells

Embryonic:Embryonic: taken from a fertilized egg, taken from a fertilized egg, about 4-5 days old. They can become any of about 4-5 days old. They can become any of your 200 cell types.your 200 cell types.

Somatic:Somatic: Found in bone marrow and the Found in bone marrow and the umbilical cord. Not as effective as embryonic umbilical cord. Not as effective as embryonic stem cells and are limited to what they can stem cells and are limited to what they can become.become.

What are the ultimate What are the ultimate goals of stem cell research.goals of stem cell research.

Uses of stem cellsUses of stem cellsMANY !!!!!!!!!!!!!!MANY !!!!!!!!!!!!!!

Possible cure for many genetic diseasesPossible cure for many genetic diseases

Alzheimer'sAlzheimer's

CancersCancers

Parkinson'sParkinson's

Limb, organ re-growthLimb, organ re-growth

Spinal cord injuriesSpinal cord injuries

Hamburgers?!Hamburgers?!

The controversyThe controversy

EmbryonicEmbryonic

Requires destruction of a human embryo Requires destruction of a human embryo (zygote)(zygote)

Recently single cell biopsies have been done Recently single cell biopsies have been done that can allow cell creation without embryo that can allow cell creation without embryo destruction! destruction!