What is Biotechnology?

Biotechnology can be defined simply as a field of applied biology that involves the use of living organisms and bioprocesses for a specific use that benefits humans. (Definition source of McGraw Hill Encyclopedia of Science). In the simplest form biotechnology is a type of biology that takes advantage of living organisms and their processes to produce materials that will improve the quality of human s life. The previous definition is accurate for defining elementary utilizations of biotechnology such as for the production of bread and alcohol however it does not describe more complex fields of biotechnology. The previous definition was credited to a source published around four years ago. Since then exponential growth has occurred in the field of biotechnology thus a new definition was created which is as follows. Biotechnology is a group of technologies that all work with living cells and their molecules for a specific use. The previous definition is much more accurate as it encompasses several aspects of biotechnology that the first definition excluded such as genetic testing, gene therapy and genetic engineering. Many people feel biotechnology is a new field of science however humans have been using microorganisms to perform certain tasks since the origins of civilization. The first known and documented use of

biotechnology is essentially agriculture as farmers for centuries have been using selective breeding to produce the most desirable crops. Two other early forms of biotechnology included the use of yeast to create alcohol and bread and the use of bacteria to create cheese.

Applications of Biotechnology
y Industrial Processes
There are two different applications of biotechnology in industry. The first is to use living organisms and their processes to simply produce a specific substance useful to industry. An example would be the production of biofuels through fermentation of sugars and starches (via various microorganisms). The second application involves using enzymes as industrial catalysts. An enzyme is essentially a protein that increases the rate of chemical reactions in living organisms. However; scientists are now using those enzymes as catalysts for industrial processes due to cheaper cost and

reduced environmental damage. An example of such is the utilization of Amylases from fungi and plants (the enzyme present in human saliva to digest food) to produce sugar from starches such as corn which in turn produces High-Fructose Corn Syrup; which is the largest type of commercial sweetener

y Medical and Pharmaceutical uses

This project focuses on thus only very brief details will be listed. The medical and pharmaceutical use of biotechnology has been the most recent innovation in the field of biotechnology. Two of the major applications of biotechnology in the medical industry are as follows. o Drug production (Biopharmaceuticals) o Gene Therapy o

y Agricultural uses
There are endless applications of biotechnology in the field of agriculture. The first and simplest applications were in the selective breeding of animals and plants to produce organisms with desirable traits. However the field has evolved to become more technologically sophisticated. Incorporating other related fields such as genetic engineering to create genetically modified organisms. The most drastic innovation in the field of agricultural biotechnology was the ability to modify organisms genetically to allow for certain desirable traits referred to as

GMO s (Genetically modified organisms). For example the technique of gene splicing (explained further on) was used on a variety of corn combining genes from soil bacteria that produces natural insecticide (Bacillus thuringiensis BT) which in the end produced a type of corn referred to as BT corn. This BT corn has the ability to produce its own toxin found in the soil bacteria that is non toxic to mammals thus no pesticides need to be added to the crop fields. This proves obvious environmental and economic benefits.

Drug Production via Biotechnology (Biopharmaceuticals)

Biopharmaceuticals are essentially just drugs made through biotechnology. In the simplest of terms biopharmaceuticals can be described as pharmaceuticals that are produced using living organisms or their active components. The most common method of creating Biopharmaceuticals is via recombinant DNA technologies (explained in Insulin Section Below). Pharming is a term used in biotechnology to describe the process of inserting desirable genes into a host organism in order for them to produce a certain pharmaceutical. Biopharmaceuticals are typically large molecules rather than synthetic drugs which typically have small molecules. The first biopharmaceutical ever developed was Humulin the term coined by the pharmaceutical company (Human Insulin). Prior to the development of Human insulin, insulin was withdrawn from the pancreas glands of either pigs or cattle.

Types of Pharmaceuticals Created via Biotechnology

y y y y Proteins Antibodies Vaccines Blood/Plasma Derived Products

Human Insulin (Biosynthetic Insulin)

The development of Human Insulin was perhaps one of the greatest medical innovations of all time. The Insulin was developed by a pharmaceutical company referred to as Genentech in 1978. Prior to biosynthetic insulin animal insulin was used as mentioned before however, on occasion our bodies developed resistance to foreign insulin; the creation of biosynthetic insulin solved that issue. The insulin was created by using recombinant DNA Technologies to insert the human insulin producing gene into the bacteria E.Coli (Escherichia coli) which in turn would produce insulin. Recombinant DNA is essentially a type of DNA that is created artificially by inserting one or more strands of DNA into a different set of DNA. Recombinant DNA is referred to as (rDNA). The process of creating Recombinant DNA to produce insulin is as shown below.

Process of Creating Recombinant DNA to produce Insulin in E.Coli (Biosynthetic Insulin)

1. DNA from one organism is extracted in order to isolate the desired gene (A gene is simply a segment of a cell s DNA that gives it a certain feature) In the case of Biosynthetic Insulin, the human insulin gene is extracted through restriction enzymes 2. Then a vector must be selected to transport the gene into another organisms (A vector is a device in which genes are transferred from one cell to another) In the case of Insulin the plasmid from the E.Coli Cell is selected (A plasmid is a DNA molecule separate and independent of the chromosomal DNA that found in bacteria as shown by figure 4) 3. DNA ligase is the sticky substance that connects exposed nucleotides so when the Insulin gene is inserted into the plasmid DNA ligase attaches them together 4. The plasmid is now inserted into the original E.Coli Cell which then reproduces several times now producing a protein that can be refined to create insulin due to the inserted human insulin gene

However; the protein produced by the transgenic E. Coli is not pure insulin it must be refined with several complex processes before suitable for human use. The method described above is just one of many possible methods of producing biosynthetic Insulin however no others are described in this project. This

process must be done on a huge scale (Trillions of Bacteria) in order to produce the amounts of insulin require to treat the approximately 2 million patients a day who use biosynthetic insulin.

Blood Clotting Factors

Thousands of males worldwide suffer from Haemophilia; a genetic disease that affects one s ability for their blood to clot or coagulate. Therefore if an injury occurs resulting in the rupture of a blood vessel your body would not produce sufficient proteins (factors) to fix the rupture. Although platelets also affect coagulation this section will simply focus on the production of proteins. There are two forms of standard haemophilia. Haemophilia A is caused by the lack of the factor eight protein, whereas Haemophilia B is caused by the lack of factor nine protein. The old method of creating these proteins for treatment was through the purification of blood. However; copious amounts of blood were needed and the blood was not adequately screened for HIV. Therefore a new method of production was required which is why CHO s were tested as a possible source. CHO s are referred to as the work horse for

biopharmaceutical companies as over 70% of all recombinant proteins produced come from CHO s. Proteins in many different forms make up the vast majority of the drugs produced through biotechnology. The reason E.Coli was not used was due to the fact that bacterial cells do not process certain human proteins thus mammalian cells were needed for further processing. The reason CHO s were used to produce the drug was essentially because they can be transfected easily (foreign DNA can be introduced into them successfully). The actual process for creating both factor VII and factor XI in CHO s is very similar to creating Insulin in E.Coli. The hardest part for the scientists was isolating the gene which produces either the factor XI or factor VII protein. Since the science of such is exceptionally complicated the process has been simplified to one step. The scientist isolated RNA of each factor then used a process called reverse transcription to transform the RNA into DNA. Once the gene was isolated from the DNA via restriction enzymes the gene was introduced into the cell via a plasmid. Since Chinese Hamsters were originally used in China in the early 20th century to perform tests with pneumococcus in the place of lab mice their cells were discovered to be excellent vectors (vehicles that deliver DNA). Therefore they can be easily transfected with plasmids. A plasmid is often the part of a bacterial cell that allows it to infect other organisms. Therefore plasmid cells were easily able to be transfected into the CHO s. Once the cell

began reproducing and depending on which gene was introduced in the original cell it would either produce factor VIII or factor IX protein. This discovery was most definitely a milestone in biotechnology as it was the first instance where a mammalian cell was used to produce a therapeutic protein.

Gene Therapy
Gene therapy as defined by the Center for Genetic Education is the use of genes as medicine involving the transfer of a therapeutic or working copy of a gene into specific cells of an individual in order to repair a faulty gene copy. Essentially that means if you have a condition that is attributed to a malfunction in a gene you insert a new functioning copy of that same gene into the cell to repair/replace it.

DNA which stands for Deoxyribonucleic acid is the genetic information contained within the nucleus of a cell that essentially controls all of the cells functions. The four bases that DNA is made from are adenine (A), cytosine (C), guanine (G) thymine (T). A gene is a segment of DNA that controls a certain feature or function. Gene therapy however is still in clinical trials for many procedures. Only one country (China) has authorized the standard use of gene therapy for combating certain types of cancer. Gene therapy is at the moment only being used on somatic (body) cells and not germ (reproductive) cells for ethical reasons. Below is an explanation of how the process of gene therapy is performed.

The Process of Gene Therapy

1. The faulty gene must be identified 2. A copy of the desired gene from the DNA of a healthy cell is obtained 3. A vector is selected to transfer the gene into the cell (Discussed at length below) 4. The gene must be properly inserted into the cell so that it is incorporated into the cells genetic makeup (Extremely complicated) 5. Once the gene is inserted into the cell it begins to manufacture or repair the protein that the malfunctioning gene was either not producing or incorrectly producing

However; the process is not as simple as it appears it is in fact extremely complicated. The hardest step for scientist in gene therapy is finding a vector to successfully deliver the gene.

Vectors Used in Gene Therapy

Harmless Viruses: A virus is basically a small infectious particle that can only reproduce and survive in the cells of living organisms. Harmless viruses are the most common vector used in gene therapy. Essentially the genetic information is removed from the virus and replaced with working human genes. The virus is then introduced to the affected cells of the human being. When the virus enters the cell membrane it is covered in a vesicle due to the cells natural defence system. When the virus reaches the nuclear membrane it injects its genetic information (the human gene) into the nucleus. If done successfully the cell should now read that genetic information and create the new protein. Both adenoviruses (viruses that contain genetic information in the form of DNA) and retroviruses (viruses that contain genetic information in the form of RNA) can be used depending on specific the virus.

Stem Cells: Stem cells are basically immature cells that have not yet been assigned a function. The desired gene is then inserted into the cell which will help it develop into a specific

cell. T That created cell will be inserted into the body at the required site and would begin to reproduce. The use of stem cells for genetic therapy is still under tested and must become more of a prevalent research topic in the field of biotechnology. Other Vectors: There are many other methods of performing gene therapy such as direct DNA transfer, particle bombardment and electroporation.