Biotech 101 is in Session …… Take your seats ………… Before we can get to the “Ah Ha” of new Biotech Research Projects, We need to learn about: –Basic Cell.

Biotech 101 is in Session …… Take your seats …………• Before we can get to the

“Ah Ha” of new Biotech Research Projects,

• We need to learn about: – Basic Cell Biology

– Basic Biochemistry

– Gene Cloning

Cells: The Basic Unit of Life

All Cells Have DNA (The “Holy Grail” of the Cell). It resides in the nucleus or nuclear region*.

A Gene is a sequence of DNA which encodes for a Trait (blood type, eye color, etc.)

nucleus

YEAST

HUMAN SKIN

BACTERIUM*PLANT

vacuole

Cells are not to scale

Plants and Animals are multicellular

Review: Basic Cell Biology

DNA

The complete set of genes is called the Genome

Proteins made from

DNA message

A-T-C-G….

The fundamental dogma of molecular biology is that genes act to create phenotypes through a flow of information from DNA to RNA to proteins, to interactions among proteins (regulatory circuits and metabolic pathways), and ultimately to phenotypes (the living being). Groups of individual phenotypes constitute a population

The fundamental dogma of molecular biology is that genes act to create phenotypes through a flow of information from DNA to RNA to proteins, to interactions among proteins (regulatory circuits and metabolic pathways), and ultimately to phenotypes (the living being). Groups of individual phenotypes constitute a population

Central Dogma of Watson & Crick:

DNADNA

RNARNA

ProteinsProteins

CircuitsCircuits

PhenotypesPhenotypes

PopulationsPopulations

Transcription

Translation

DNA is like videotape

Proteins: “products made from the

blueprint on the DNA” • Essential chemicals for cell structure and activities (reproduction, moving around, defense against invaders, metabolism, etc.). Insulin is a protein.

• Basic unit: amino acid (~20 different ones) An average protein has ~500 amino acids.

• The arrangement of these amino acids determine the type of protein made

• Codons (triplet code of bases) on mRNA determines the sequence. “Universal Code”– AUG-UUU-... --> methionine-phenylalanine-...

Gene Cloning: 4 essential steps:

1. Obtain donor DNA = (promoter (start code)+GOI*+stop code + marker gene). *gene of interest

2. Link donor DNA to vector DNA (such as plasmid or phage) via restriction enzymes (“scissors”) & DNA ligase (“glue”)

3. Insertion of the recombinant DNA (rDNA) into a host cell such as bacteria, yeast, plant or animal. “Transformation”

4. Detect recombinant clone (transgenic organism) or new gene product (protein) [marker gene helps to identify the transformed cells]

Clone = Identical Twin

Plasmid =

Small circular DNA found in bacteria and

yeast

EcoRI = DNA-

cutting enzyme

from E.coli

Example: Insulin gene

A Few Applications of Biotechnology:

• More nutritious crops with longer shelf lives. • Higher yields per acre due to resistance to pests,

herbicides and environmental conditions such as drought, salinity, etc.

• New & Safer vaccines and drugs, esp. for cancer • Transgenic animals & plants which produce human

hormones, antibodies or enzymes.• Bioremediation to clean up pollutants by altering

microbes or plants.• Therapeutic cloning: example: grow skin cells for burn

victims

Genetic Engineering enhances the process of Conventional Selective Breeding: It is faster, more precise and doesn’t require sexual compatibility

Salt Tolerant Tomatoes May Hold the Key to Growing Crops in Marginal Soils (and may even reclaim salt from the topsoil)

• 24.7 million acres of once agriculturally productive land are being lost annually because of irrigation-induced salinity (USDA). • Crop production is limited by salinity on 40 % of the world's irrigated land and on 25 % of the land in the USA. • Blumwald and Zhang genetically engineered tomato plants that produce higher levels of a “sodium transport protein." Store salt in vacuole of leaves/roots• Plants grow and produce fruit even in irrigation water that is > 50X saltier than normal. Nature Biotechnology, Aug 2001

Blumwald & Zhang’s Work

Transgenic tomatoes in 5mM (A) and 200mM (B) of NaCl

2 & 3 Have gene for transporter

UC Davis Research:

“Pharming” represents the Third Wave of Ag Biotech

• Use of Genetically Modified Plants or Livestock as factories, rather than food

• Plant derived medicines is not new! Examples: Taxol, digitalis, quinine, etc.

• A plant/seed can be a high protein expression system (ex: Ventria Biosciences in Sacto)

• Proteins can also be expressed in milk, semen or urine of cows, pigs, rabbits, etc. Example: spider silk (Nexia)

• Significantly less costly than stainless steel tanks due to low initial captital investment and scalability

Nature Biotechnology

Uncorking the biomanufacturing bottleneck

Alan Dove, Aug 2002 p777-9• “As biomanufacturing capacity becomes strained, several new methods

for producing biologics are being investigated by biotechnology companies”

• Spokesperson from GTC Biotherapeutics (formerly Genzyme Transgenics) in Framingham, MA estimates a 200% increase in manufacturing capacity for Mabs alone over the next 10 years. Their solution is milking transgenic animals (“mammary bioreactors”). PPL Therapeutics (UK) and BioProtein (France) have similar projects. Goats, sheep and cows are the most common mammals.

• Transgenic chicken eggs may also be feasible, but the research is still in early stages. Companies: Origen Therapeutics (Burlingame, CA); Avigenics (Athens, Ga); TranXenGen (Shrewsbury, MA); GeneWorks (Ann Arbor, MI) and Vivalis (France).

Use Tobacco to make medicines

Large Scale Biology (Aka Biosource) in Vacaville, Ca is using the tobacco plant as a production system for human vaccines, cancer therapies (ex: lymphoma) and other pharmaceuticals. “Pharming in Plants” but no gene flow!

(Transient expression of mRNA of vaccine gene)

“Personalized medicine”

RNA

www.lsbc.com

Over 120 million children worldwide are deficient in vitamin A. Rice has been engineered to accumulate -carotene, which is converted to vitamin A in the body. Incorporation of this trait into rice cultivars and widespread distribution could prevent 1 to 2 million deaths each year.

Increased -Carotene in Rice Grains

(bacteria)

(daffodil)

Normal rice

“Golden” rice

(daffodil)

Introduced enzyme(source)

Ye et al. (2000) Science 287: 303-305.

Normal rice

Edible VaccinesIntroduce antigenic proteins from disease-causing organisms into plants. Eating the fruit or vegetable can then induce antibodies just like a vaccination, rendering the person immune to the disease.

The feasibility of this approach has already been demonstrated. Dr. Charles Arntzen of Arizona State University. He is actively pursuing research to allow children to be immunized against debilitating diseases such as hepatitis B, for example, by simply eating a modified banana, potato or tomato. Also Bill Langridge, Loma Linda Univ. in California. ( Sci. Am., Sept. 2000)

Nexia's lead products are biomaterials and pharmaceuticals used to treat life-threatening disease. Nexia produces these recombinant proteins in the milk of transgenic BELE® goats from which they are extracted. Nexia will further process the milk to purify recombinant products. BioSteel™ filaments are environmentally friendly and will be used in applications where strength and flexibility are required, such as medical devices or body armor.

Canada

Who Assures Safety of Biotech ?

Agency Products Regulated

• U.S. Department of plant pests, plants, veterinary Agriculture (USDA) biologics, animals, fish

(Safe to Plant)• Environmental microbial/plant pesticides, Protection

new uses of existing Agency (EPA) pesticides, novel

microorganisms (Safe for the Environment)• Food and Drug food, feed, food additives, Administration

veterinary drugs, human (FDA)drugs and medical devices

(Safe to Eat)

Web Sites for Information:• To see a weekly summary of world developments in agri-biotech , produced by the Global

Knowledge Center on Crop Biotechnology, International Service for the Acquisition of Agri-biotech Applications (ISAAA), and CAB International. (http://www.isaaa.org)

• Essential Biosafety CD: http://www.essentialbiosafety.info (free). This CD-ROM offers comprehensive information about the safety of genetically modified crops. Second edition will be released in Nov, 2002.

• Food Biotechnology: A Communication Guide to Improving Understanding is available! This Guide aims to raise awareness of this communications issue, and provides lots of practical advice and resources to overcome this problem.. Get your free CD by e-mailing a request to [email protected]. 

We have been here before!

We have recently advanced our knowledge of genetics to the point where we can

manipulate life in a way never intended by nature.

We must proceed with the utmost caution in the application of this new found

knowledge.LUTHER BURBANK, 1906