What is gene cloning and how does it work?
DNA cloning is the process of creating multiple identical copies of a piece of DNA extracted from an organism. The very first step of making insulin is the synthetic creation of this human insulin gene. The human insulin gene is at the top of chromosome 11 in humans. Firstly there is a double-stranded DNA, if a part of this DNA has a gene that we would like to clone, the first thing that has to be done is to cut the gene out, this is done by restriction enzymes, which are found naturally in bacteria. Restriction enzymes attach onto the DNA strand and identifies the genetic sequence and ‘cuts’ it in the right place. Now you will be left with the gene. Now we paste this gene into a plasmid using another enzyme called DNA ligase. This then forms a recombinant plasmid that contains our desired gene that we wanted to clone. A plasmid is a piece of genetic material that sits outside of chromosomes that are capable of replicating identically. Once we have “cut” out our gene we then “paste” it into a plasmid using a process called transformation, this plasmid is now genetically modified. Now we have a plasmid which we then insert it into a bacterial cell. Once the plasmid is “pasted” into the bacteria, the bacteria will now be able to survive and reproduce identical copies of bacteria that also contains the new gene and plasmid this is done by a process called binary fission.
What are the possible uses for genetic cloning?
There are three different types of artificial cloning which are: gene cloning, reproductive cloning and therapeutic cloning. One of the main reasons for cloning genes is to gain information about the nucleotide sequence of a gene. Another main reason to why someone would want to gene clone is to manipulate a gene. Meaning to change the gene’s DNA sequence or to merge different DNA molecules together. Cloning genes can help cure many genetic disorders such as cystic fibrosis and immunodeficiency (SCID).When plasmids are created to carry a normal version of a gene that is nonfunctional in cystic fibrosis, these plasmids can help the functioning of patients lungs who have cystic fibrosis. Plasmids are delivered to the lungs of cystic fibrosis patients, lung function becomes better and improves the ability of airway secretions to kill bacteria. Helping to cure these disorders with the help of gene cloning make huge impacts on people's lives. Gene cloning for medical purposes helps a lot of people. Cloning can also be used to clone different species which have become extinct. When gene cloning is done it can be useful for many different processes, For e.g, detection of diseases, replacement of defective genes. Over many many years gene cloning has been beneficial for scientists to further investigate the genes that are inside our bodies.
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What is type 1 diabetes & Insulin?
Diabetes Mellitus type 1 is a condition in which the pancreas which is located behind the stomach, produces little or no insulin. Insulin is a hormone made by the pancreas. Inside your pancreas there are small clusters of cells called islets of Langerhans more commonly known as islets. People who have type 1 diabetes immune system attacks the insulin producing cells therefore damaging them and stopping the production of insulin, which your body needs.The main purpose of insulin is to lower blood sugar known as glucose for energy. Eating causes blood glucose levels to rise. Cells (known as beta cells) are released from your pancreas and signaled to produce insulin into your body. Insulin allows cells to absorb glucose allowing it to be used as energy. Insulin helps to remove sugar from the blood. Insulin also helps to store sugar in the liver and muscles and releases it when your body needs it for e.g, during exercise. When your body does not produce enough Insulin, cells are not able to absorb glucose and used for energy, as a result your body breaks down your fat and protein stores as an alternative way to provide energy. When your body can not produce enough insulin this causes high glucose levels which can cause long-term complications if the blood sugar levels stay high for too long. For a person who has type 1 diabetes it is important for them to keep their glucose levels at a healthy range. By taking insulin as well as eating a healthy diet and exercising regularly you can keep your blood glucose levels within a healthy range. The exact cause of type 1 diabetes is unknown, but in most people with type 1 diabetes, the body’s immune system which normally fights harmful bacteria and viruses mistakenly destroys the insulin producing cells in the pancreas.
Link between genes, chromosomes, protein, insulin, how do bacteria grow, make protein
A gene is a small section of DNA which codes for a specific protein. Genes contain information that our bodies need to make chemical’s called proteins. So, for e.g one gene will code for the protein insulin,which has an important role in controlling glucose levels in your blood. The INS (Insulin) gene is a protein coding gene. This gene provides us with instructions on how to produce the hormone Insulin, which is needed to control glucose levels in the blood. The human insulin gene is at the top of chromosome 11 in humans. Bacteria are tiny living single-celled organisms. Bacteria is found everywhere, from inside your body to everywhere around you. If bacteria are in the right conditions like moisture, nutrition, pH and temperature then doubling of bacteria can occur quickly. Each bacterium has its own condition that it chooses to grow in. Under the right conditions some kinds of bacteria can double every 20 minutes. Bacteria reproduce very simply and fast, by dividing and splitting into two, two becomes four, and the dividing process continues. DNA codes for proteins, the cell sends enzymes to copy parts of the DNA that codes for an individual protein. That’s then transported out into the cell, where it is paired with a ribosome, and translated into a protein. The first protein drug to be produced commercially in bacteria was insulin. To produce a protein in bacteria, “you first need to clone the gene that encodes it, then introduce the vector containing your gene into bacteria”. It is very important to make sure that the vector you clone it into is an ‘expression vector’. By including this bacterial promoter, you are giving the bacterium orders to make a protein from your gene of interest. You are basically ‘tricking’ the bacteria into creating a foreign protein. In today's society bacteria and other organisms are used as biological ‘factories’ to make protein medicines in large amounts, this is one of the easiest ways to produce a lot of your desired protein.
Economic advantages and disadvantages:
Animal Insulin, how is it made, how much does it cost?
Animal insulin was the first type of insulin to be administered to humans to control diabetes. Animal insulin is taken from the pancreas of animals, commonly from pigs (porcine or pork insulin) and cows (bovine or beef insulin). The discovery of insulin happened in 1921, by a Canadian surgeon named Frederick G. Banting, his chemistry assistant named Charles Best, and John MacLeod. In 1921, Banting and best removed a pancreas of a dog then they “ligated” the pancreas of another, stopping its nourishment. After removing the pancreas from a dog they then threw the pancreas into a blender with some saline ice. And then reinjected the dog which now has diabetes, with its own blended up pancreas.The procedure worked. The dog did not die straight away, it recovered some of its strength. These two men discovered that injecting the dog a few times a day kept the dog alive and healthy. Betram Collip later on began helping these men, and they also started using bigger pancreas’ found in cows. Collip helped to purify the insulin to be used for testing on humans. Banting and Best were confident of their discovery of Insulin and decided to test their findings on themselves. After their testing had been done, their Insulin was ready to be used on patients. After testing the insulin of different patients it proved to be a success, and Banting and MacLeod were awarded the Nobel Prize in Physiology or Medicine.
The first “sale” of insulin was just for $3 (Canadian), not for a vial of Insulin but for the intellectual property of the drug. For $1 the researchers assigned their patent rights to the University of Toronto. In 1941 it was reported by Mr. Eli Lilly that Insulin cost 7.5 cents per day for diabetics. A bottle of pork insulin in 1972 cost $1.49. In Atlanta in 1974-1976, a couple bottles of insulin (NPH beef and pork) costed $2.50. It cost $20-25 for a month of insulin supplies. One vial might last a patient less than two weeks.
Nowadays the price of insulin has increased significantly. Rapid acting insulin can be anywhere from $93 for a vial of Novolin R, to $275 per vial of Humalog. That is a significant increase from a vial of Insulin for a cost of $1.49. People with diabetes can require 1-6 vials of insulin per month.
6Nowadays the price of insulin has increased significantly. Rapid acting insulin can be anywhere from $93 for a vial of Novolin R, to $275 per vial of Humalog. That is a significant increase from a vial of porcine insulin Insulin for a cost of $1.49 in 1972 . People with diabetes can require 1-6 vials of insulin per month.
Production costs: There are different costs between producing gene cloned insulin and animal insulin, because they are both made differently. Manufacturers have to buy raw materials, pay workers, and buy equipment. A paper produced in 1995 focused on the commercial production of insulin using E.coli bacteria. Researchers found that the total cost in making enough insulin to treat one patient a year costed $33.60. Most of this cost is based around the steps of purification of insulin and the fermentation process. Production prices have increased since this paper was recorded due to the new DNA technology in our society, the development of more and different equipment, and the public’s demand for genetically produced insulin. A 2018 study on the yearly costs for the production of insulin reported that the estimated biosimilar prices were from $48-$71 per patient per year.
When the first kind of human insulin hit the market in 1982 animal insulin production declined. With the creation of gene cloned insulin took away any concerns about transferring any possible animal diseases into the insulin. There are now very few companies which still sell animal insulin one of them being ‘Wockhardt UK’ a company which sold porcine and bovine insulin for many years. In 2017, ‘Wockhardt UK’ decided to discontinue the production of bovine insulin due to “global bovine insulin raw material unavailability, the company has been left with no choice but to discontinue the Hypurin® Bovine insulin range”. This company still sells porcine insulin to this day and many years to come but just not bovine insulin. Due to this company’s unavailability of raw materials to make it, the cost of production to make animal insulin would have increased, due to the rising demand of cow pancreas’. Due to the inaccessible resources, insulin companies would of wanted to change to producing insulin from gene cloning. Compared to gene cloned insulin, animal insulin would of had a higher cost of production due to the demand in resources.
During the days of creating pork insulin, it took more than 2 tons of pig parts to produce 8 ounces of purified insulin. This goes to show the very small amount of animal insulin we get out of such a large amount of animals. Breeding and raising of livestock also have to be considered when looking at the total production costs of making animal insulin. Eli Lilly (past manufacturer of animal insulin) needed 56 million animals per year to meet the USA’s demand for the drug. With the help of microorganisms we use to create gene cloned insulin today provides more of the hormone. Bacteria multiplies at rapid speed, so for producers it is easier to make insulin from gene cloning than animal insulin. Without the discovery of gene cloning to produce insulin, animal insulin resources would definitely not have met the public's demand for insulin.
Everybody’s preference and needs for which Insulin they use varies. One of the most crucial parts in picking which insulin to use for many people would be price which would be a massive factor for some. For one vial of insulin can cost from anywhere between $25 to $417, (depending on if you buy standard, short acting or rapid acting insulin). For many people around the world people can't afford insulin. Many families whose children suffer from diabetes find it hard to find the funds for insulin. Some families have even had to create GoFundMe pages to fundraise for the medicines. There are even some reports where patients were rationing insulin, and in some cases even dying because they couldn’t afford the drugs they needed to survive. This just goes to show how expensive these medicines really are. People in developing countries are deprived of these medicines. It is proven that gene cloned insulin can be double the amount of animal insulin or even triple for some kinds of gene cloned insulin. For a standard vial of porcine insulin it can cost anywhere from $60 and $90. Even though this is slightly cheaper than gene cloned insulin, there are still many additional charges that come with animal insulin.
In 1998 the manufacturing of beef insulin for human use was discontinued in the USA. In 2006 the manufacturing of porcine insulin for human use was discontinued in the USA. USA has 9.4% of the world's population who have diabetes, many of these people need to buy insulin. Animal insulin is a lot harder to attain. For people in places like the USA they have to import their animal insulin, and it then has to get checked by the FDA and they have to decide if they will let it into the country.This would result in extra shipping and handling fees, and there is no guarantee that a mistake could happen during the journey. Gene cloned insulin is much easier to get your hands on and a lot more popular, according to the Eli Lilly corporation, “in 2001, 95% of insulin users in most parts of the world take some form of human insulin”, this shows that human insulin would be a more popular option for many people. Gene cloned insulin is obtainable through a prescription from your doctor. Gene cloned insulin can even be brought from a supermarket called Walmart. For some patients, it would be easier to buy gene cloned insulin over animal insulin even though gene cloned insulin is proved to be more expensive. Synthetically made insulin is proven to be easier because the colonies of bacteria are produced at such a rapid speed, meaning there is less resources needed to make insulin and more of the desired insulin, this way in which compared with insulin made from pig and cow pancreas’ need much more time, effort and resources involved with the process, and the result is a small amount of insulin for how many resources would be used.
Others P.O.V’s of using gene cloning to produce insulin
Everyone has different thoughts when it comes to the idea of using gene cloning to produce insulin. Some people think that using gene cloning to produce insulin goes against the laws of nature, or against people’s religious beliefs. Many others are mostly concerned about safety issues. Using gene cloning can also be beneficial to people is so many ways, usually people in the past were only expected to live 4 more years after being diagnosed with diabetes, but now that we have the new DNA technology to produce human insulin, humans can be expected to live a normal life, with a normal life expectancy. Many people see it as a great and effective way to improve people's lives.
PROS:
12Sverker Johansson is a Swedish Physicist who sees genetic engineering as a sign of progress and an opportunity to make the world and the lives of humans and animals better. He also believes that genetic engineering can make pharmaceutical products more effective. “There are already improved versions of insulin for diabetics and human growth hormones on the market thanks to genetic engineering”. Sverker finds all the positives of genetic engineering, from making fruits even bigger and better than the ones before to improving the lives of so many individuals because of genetic engineering to produce insulin, 'I believe that genetic engineering, can benefit people in several ways, for example, scientists made artificial human insulin with the help of recombinant DNA technology”.
CONS:
The Catholic Church’s views on genetic engineering: There are many different responses to genetic engineering in the catholic faith:
One of the reasons would be ‘anti-god’. Many Christians believe that God created all living beings on earth (Colossians 1:16), and by genetically modifying creatures we are going against the authority of God. “The belief that God should have ultimate power and we should not be altering nature is what some Christians believe should have us halt the progression of human GE”. Most of the people in the Catholic Church agree with gene therapy to treat diseases but not therapeutic cloning. They believe that “without certain diseases, it would be possible for more people to live longer, healthier lives”. Gene therapy is used to cure many different diseases, the Catholic Church agrees with the use of gene therapy but not so much with therapeutic cloning (human cloning).
(Statement from Pope John Paul II) “Reproductive and therapeutic cloning seems specious. Both forms of cloning involve disrespect for the dignity of the human being. In fact, from an ethical and anthropological standpoint, so-called therapeutic cloning, creating human embryos with the intention of destroying them, even if undertaken with the goal of possibly helping sick patients in the future, seems very clearly incompatible with respect for the dignity of the human being, making one human life nothing more than the instrument of another”. The Pope believes that God is the one who hand crafted our souls from the moment of fertilisation, therapeutic cloning goes entirely against this. Christians think that the dignity of the person is being overlooked.
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