BiotechnologyBiotechnology

Biotechnology Biotechnology Definition:Definition:

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

Recombinant DNARecombinant DNAGenes from different species are combined Genes from different species are combined together and placed into cellstogether and placed into cells

The 2 samples are combined and make a hybrid The 2 samples are combined and make a hybrid DNADNA

Hybrid DNA gets inserted into bacteria which Hybrid DNA gets inserted into bacteria which multiply with the recombinant DNA insidemultiply with the recombinant DNA inside

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

Making Recombinant DNAMaking Recombinant DNA Let’s use the example of Green Fluorescent Let’s use the example of Green Fluorescent

Protein (GFP). It occurs naturally in jellyfish Protein (GFP). It occurs naturally in jellyfish and fireflies, but we want to put it into the and fireflies, but we want to put it into the genome of a cat.genome of a cat.

1.1.Isolation of the gene of interestIsolation of the gene of interest• ““Cut” the GFP gene from a Cut” the GFP gene from a

jellyfish’s DNA using a jellyfish’s DNA using a restriction enzymerestriction enzyme

2. Insertion of the gene of 2. Insertion of the gene of interest into a plasmid vectorinterest into a plasmid vector• Vector is a small, circular Vector is a small, circular piece of DNA (plasmid)piece of DNA (plasmid)•GFP gene + vector = hybrid GFP gene + vector = hybrid PLASMIDPLASMID

3. Transfer of the plasmid into the 3. Transfer of the plasmid into the organism to be modifiedorganism to be modified• Plasmid gets inserted into EMBRYONIC cat Plasmid gets inserted into EMBRYONIC cat

cellscells

Cat’s Chromosomes within nucleus

Plasmid

44. Transformation of host . Transformation of host organism’s cellsorganism’s cells•Over time, the plasmid containing Over time, the plasmid containing the gene for GFP will be the gene for GFP will be incorporated into the cat’s DNAincorporated into the cat’s DNA•This new gene will change the This new gene will change the proteins producedproteins produced

• 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

1.1. PharmaceuticalsPharmaceuticals: : HGH, Insulin: insertion of HGH, Insulin: insertion of human gene into bacteria for cheap proteins.human gene into bacteria for cheap proteins.

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

http://www.abpischools.org.uk/page/modules/diabetes/diabetes6.cfm?coSiteNavigation_allTopic=1

2.2. AgricultureAgriculture::

-Crops: Disease protection, drought & frost -Crops: Disease protection, drought & frost resistance, increased growthresistance, increased growth

-Livestock: Increased muscle mass, hormone -Livestock: Increased muscle mass, hormone acceleration, drugs in milkacceleration, drugs in milk

Let’s take a closer Let’s take a closer look…look…

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.

How do they genetically How do they genetically modify foods?modify foods?

Recombinant DNA!Recombinant DNA!Desired genes are transferred to specific crops Desired genes are transferred to specific crops

Examples: bigger size, specific nutrients, resistance to Examples: bigger size, specific nutrients, resistance to drought or pestsdrought or pests

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

-GM crops can be engineered to grow faster.

-GM crops can be engineered to be pest-resistant

-GM crops can be engineered to tolerate extreme weather conditions

-GM crops can be engineered with added vitamins and minerals, which is especially beneficial in third world countries dealing with malnutrition

ConsConsThere are a lot of unanswered questionsThere are a lot of unanswered questions

What are the long term health What are the long term health effects?effects?Can GM foods cause new allergies?Can GM foods cause new 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?

We do not fully understand the effects GM food We do not fully understand the effects GM food can have on our worldcan have on our world

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 ingredientsSoy lecithinSoy lecithin

FructoseFructose

DextroseDextrose

Rapeseed oilRapeseed 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….

Crime Scene Crime Scene InvestigationInvestigation

DNA FingerprintingDNA Fingerprinting

• Each person has their own unique Each person has their own unique DNA DNA “fingerprint” (specific nucleotide sequence)“fingerprint” (specific nucleotide sequence) except for identical twinsexcept for identical twins

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

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

Gel Electrophoresis 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 How does gel electrophoresis work?electrophoresis work?

Restriction enzymes Restriction enzymes cut DNAcut DNA

Pieces of DNA are run through a gel Pieces of DNA are run through a gel electrophoresis tray electrophoresis tray

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

Only about 10 regions are testedOnly about 10 regions are tested

The probability of having matching The probability of having matching fingerprints is ~1 in 1 millionfingerprints is ~1 in 1 million

Gel ElectrophoresisGel Electrophoresis

A case for MauryA case for Maury

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

• As is to be expected, the As is to be expected, the mother and the child share mother and the child share some DNA bands since they some DNA bands since they share 50% of their genes. share 50% of their genes. However, they each have However, they each have bands that are not bands that are not represented among the bands represented among the bands of the other. of the other.

• What do you think - is What do you think - 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 DNA Shown to the right are DNA "fingerprints" taken from "fingerprints" taken from seven (7) suspects (the seven (7) suspects (the numbered columns). numbered columns).

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

Endangered SpeciesEndangered SpeciesDNA can also be used to catalog DNA can also be used to catalog endangered species endangered species

Endangered Birds: Collect feathers Endangered Birds: Collect feathers to identify DNA instead of trying to to identify DNA instead of trying to capture and mark every single birdcapture and mark every single bird

In most cases when DNA In most cases when DNA fingerprinting was used they got a fingerprinting was used they got a higher number of species than higher number of species than when models were usedwhen models were used

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 ProjectAimed to sequence Aimed to sequence every single every single gene in gene in humans!humans!

1990-20031990-2003

Goals:Goals:Identify ALL genes in human DNAIdentify ALL genes in human DNA

Determine sequence of the Determine sequence of the 3 billion 3 billion base pairsbase pairs

Store all information in databaseStore all information in database

Improve tools for analysisImprove tools for analysis

Transfer information to the private sectorTransfer information to the private sector

Address all ethical and legal issues Address all ethical and legal issues

Implications???? Implications???? http://www.youtube.com/watch?v=F5LzKupeHtw

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 : http://www.youtube.com/watch?v=kp1bZEUgqVI

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

using using biotechnology??biotechnology??

…Yes! Its called …Yes! Its called gene therapygene therapy!!

Gene TherapyGene Therapy

• Replacement of defective genes with healthy Replacement of defective genes with healthy genesgenes

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

sickle-cell anemia, immune-deficienciessickle-cell anemia, immune-deficiencies

Gene Therapy: Cystic Gene Therapy: Cystic Fibrosis Fibrosis

Cystic fibrosis Cystic fibrosis is caused by a defective is caused by a defective gene that causes humans to produce gene that causes humans to produce excessive amounts of mucusexcessive amounts of mucus

This disease is life threatening This disease is life threatening

Gene therapy Gene therapy replacesreplaces that defective that defective gene with a normal gene which causes gene with a normal gene which causes the over production of mucus to stopthe over production of mucus to stop

CloningCloning

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

Bacteria and sponges often reproduce by Bacteria and sponges often reproduce by cloningcloning

Identical twins are natural clones (but not Identical twins are natural clones (but not fraternal – why?)fraternal – why?)

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

Egg cellPin

Needle

How it works…How it works…

The nucleus of an egg cell is removed and replaced The nucleus of an egg cell is removed and replaced with a nucleus taken from an adult organismwith a nucleus taken from an adult organism

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

The nucleus from the adult organism will be The nucleus from the adult organism will be identical to the offspring bornidentical to the offspring born

http://learn.genetics.utah.edu/content/tech/cloning/whatiscloning/scnt.html

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

People already eating food from cloned People already eating food from cloned animalsanimals

United States bans human cloning of all types United States bans human 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.

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!

Latest researchLatest researchIt is possible to create a stem cell using adult cells It is possible to create a stem cell using adult cells (no embryos needed) but the down side is that (no embryos needed) but the down side is that these adult cells are more likely to form tumors.these adult cells are more likely to form tumors.

Finding new stem cells in amniotic fluid could be Finding new stem cells in amniotic fluid could be an alternative to embryonic stem cellsan alternative to embryonic stem cells

In March 2009, President Obama lifted In March 2009, President Obama lifted restrictions on stem cell research. This will allow restrictions on stem cell research. This will allow scientists to conduct research on already-existing scientists to conduct research on already-existing stem cell lines.stem cell lines.

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

Uses of stem cellsUses of stem cells

MANY !!!!!!!!!!!!!!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