IMMORTALITY Genetic Engineering and Genetic Therapy
Genetic engineering and computer coding are broadly similar, if someone has a "cassette" or "programme", a biologist or biochemist would have coded the cassette to function and other people can look at the cassette or code as open source, upload online, modify, use it and improve on it, be it lactose code or spicy tomatoes, giant strawberries and as far as the imagination can go.
Gene therapy is a procedure to change a gene or a group of genes to cure some medical problem. Theoretically it is possible to treat disease and to add ideal traits such as improved vision, maximum muscle density, perhaps regenerating new teeth in old age, egotistical, cosmetic reasons and more. Biologists have mapped many genetic disorders to their chromosome and gene. The genome code is long, reads like a dictionary without words to definitions meta structure, and there are more mysteries yet to be uncovered. Our study lead us to DNA code because we find that there is always a deeper cause, and we are led deeper and deeper in the search for an answer. Testosterone or any protein is produced by the pituitary gland, and with aging the body produces less testosterone. The blueprint for making testosterone is in the DNA, epigenetics sits on top of DNA and turns genes on and off and organelles in the cells manufacture the proteins and send it where it needs to go. All must be optimal for overall health in an individual. Genetic engineering is about rectifying gene errors or mutations and configuring and controlling the epigenome, controlling methylation and demethylation targets and landscapes.
Human beings are at the stage of chemistry. When atoms share electrons they become chemicals. Different atoms or elements in chains, molecules, polymers, compounds, acids. Just like magnets prefer opposite poles to like poles or an electron following the power shell rules, complementary chemistry preferable chemical bonds form a natural affinity in the building and maintenance of the human body. If convection operates on radiating from hot to cold then you could set a temperature for the desired effect, if you have too much water you could raise the temperature, too little water and you could lower the temperature, conduction. One chemical will prefer a specific chemical more so than just any chemical and these little rules are utilized so that human body is naturally complementary. Oil is not soluble in water so it is utilized to form a cell that holds water and so on. Acids are used in the human body because they are an unstable atom that can force in an electron or send out a proton, by doing so it can work to design and redesign to build and maintain the human body, a stable atom that does not pull or push is not capable of being a chemical building block. (De-oxy-ribo-nucleic-acid) DNA, Ribonucleic acid (RNA), amino acids, vitamin C known as ascorbic acid, also trace elements such as iron and potassium. Molecules form when two or more atoms form chemical bonds with each other. Nucleotides are any of several compounds that consist of a ribose or deoxyribose sugar joined to a purine or pyrimidine base and to a phosphate group and that are the basic structural units of nucleic acids (such as RNA and DNA) — compare nucleoside. A nucleoside consists of a nitrogenous base covalently attached to a sugar (ribose or deoxyribose) but without the phosphate group. A nucleotide consists of a nitrogenous base, a sugar (ribose or deoxyribose) and one to three phosphate groups. The elements that form DNA are Hydrogen, Oxygen, Nitrogen, Carbon, Phosphorus.
The human body is made of cells, when you are born it is one cell that divides so most of your body is made of cells, the extra cellular matrix is the scaffold that glues all the cells together. There are two major roles of the cell, one is called mitosis, the cell copies itself and then divides multiplying and secondly, making proteins. Proteins are a general term for any substance cells manufacture (est. 20,000 to over 100,000 proteins). It is important to observe the entire cell not just the DNA because many pathologies are cell degradation related. The cell has little organs called organelles like the organs in the body perform their particular task. Cells organize many of their biochemical reactions in non-membrane compartments. Recent evidence has shown that many of these compartments are liquids that form by phase separation from the cytoplasm.
The instructions to build the human body is called DNA (De-oxy-ribo-nucleic-acid) and every cell in the body has its own copy of the entire DNA, in the nucleus of the cell is the chromatin or commonly referred to as DNA, your own version of the human genome. Everyone always says DNA is a double helix, flatten it out it looks like a ladder, the steps of the ladder are called base pairs, they are repeating letters that form the code C, G, A, T, they are the first letters of amino acids. C stands for Cytosine, G for Guanine, A for Adenine, T for Thymine, and they are instructions for building proteins (a bunch of amino acids in a group is called a protein). A group combination of these amino acids are for example sulphuric acid, insulin, testosterone and so on. The cell receives a signal, unwraps the DNA, reads the DNA and makes a specific protein, for example we drink milk the body detects lactose and makes a protein to break down lactose into sugar, or the pancreas cell senses blood sugar, reads the DNA and makes insulin and sends it into the blood stream or the pituitary gland makes a hormone.
DNA is the master copy while RNA (Ribo-nucleic-acid) is the carbon copy or the working template. RNA has the letters C stands for Cystocine, G for Guanine, A for Adenine and U for Uracil replaces Thymine in DNA. When a signal is received to make a protein called transcription factors the DNA which is wrapped in histones, those wrapped tightly are rarely used while those wrapped loosely are commonly used, unwraps the DNA and opens it up so that RNA can attach to the base pairs and build a copy. Once RNA has read the DNA is wrapped up again but the RNA undergoes further processing to remove some extra pieces and caps are placed on the ends then it floats out of the nucleus and into the endoplasmic reticulum where the RNA is read three letters at a time or in triplets and a corresponding amino acid is constructed, after which we have something called a peptide. The peptide is sent to the Golgi apparatus to be folded into a protein and finally sent into the bloodstream.
There are checks and balances to ensure no mistakes are made, for example DNA has a 5 carbon 3 carbon orientation so the RNA reads in the correct direction. Letters have chemical affinity and correspond strictly with another letter, G DNA always links with C RNA, C with G, A with U and T with A. There are start and stop genes.
In genetic engineering being able to make proteins or new cells is very powerful. Generally we are seeking to do two tasks. One is to change the code, either 1.) adding code, 2.) subtracting code or 3.) changing existing code. The code we want to make usually has some aim, so we start with an aim, this commonly means curing some medical issue or making a protein or molecule that is of some upgrade. We can compare traits in different people to determine what the code needs to be, or we can take some relevant chemical snippets and determine if or how the cell can make that substance. Genetic engineering is sought to be done in vivo or in vitro. In vivo, making alterations to a living human. Each and every cell in the human body has its own copy of the complete DNA so if you seek to change some DNA it has to be done to the relevant part of the DNA and either localized to the specific part of the body or to the entire body. If you make a DNA change to the part of the genome relating to the eyeball and it needs to be placed with the part of the genome relating to eyeballs otherwise your toes may start growing retinas and you do not want that. In vitro means the cells are altered in a lab and never installed in a human, for research, Ex In vitro or Ex Vitro are made in a lab and intended to be injected into the human body. Delivery is an ongoing technological issue of gene therapy. Ideally we would want infection by phenotype and not adding the same code again with each infection. Safety is paramount.
Secondly, ideally we want the option to delete the alteration reverting before the gene therapy allowing us to either end or upgrade and gene expression and regulation to be able to turn our code on or off. An embryo goes from a lump of cells to a human being, the lump of cells differentiates into specific body parts, so it no longer looks like a ball of cells but a human being. The cell differentiates so there are skin cells, liver cells, pancreas cells, brain cells, fat cells, nerve cells and so on but each and every cell contains a complete copy of the DNA, a pancreas does not make sulphuric acid and the stomach does not make insulin. This is called the cell's phenotype. A cell of a particular phenotype could theoretically be a cell of a different phenotype because it contains all the DNA blueprint, a hair cell could be a liver cell if the genes for cell expression of hair were turned off and the genes for cell expression for a liver cell were turned on, by expressing some genes and not expressing other genes the cell has an identity. This is called epigenetic editing. There are over 200 different cell types in the human body. Our code or method should have on and off switching ability.
If we want to add our own genetic code to the human cell, we want our own little off and on switch. If a person is lactose intolerant for example he does not want his genetic engineered cells to process lactose all the time, no he will only want to process lactose when milk is present. This acts like an on and off switch with the milk being the trigger.
Oh, and attempting to understand the code of course, what each part does is required for working the code. All of these problems require intelligence on the part of the genetic engineer to overcome. Scientists may knock out a gene in a mouse or bacteria and then test to see the difference, if they knock a random gene and the bacteria stops making some chemical then it uncovers what the gene is required for. A gene chip, by taking a sample, one can see what genes are being expressed in humans and what genes are not being expressed. A key tool in possibly determining what genes are responsible. For example, you can get an old person and a young person and see the differences. A whole group of genes may form the expression to a particular trait rather than just one gene.
So three technologies rather than two form the competency of a genetic engineer.
Altering Bacteria DNA In The Human Body Instead Of The Body Itself
There are many bacteria colonies in the human body that have their own DNA. We can in vitro genetic engineer these bacteria to do some task, drink the kefir, and we also have the option to remove them by taking antibiotics or trigger some self-destruct gene sequence. Organisms are known to live on the skin, mammary glands, placenta, seminal fluid, uterus, ovarian follicles, lung, saliva, oral mucosa, conjunctiva, biliary tract, and gastrointestinal tract. Types of human microbiota include bacteria, archaea, fungi, protists and viruses.
We want them to be able to manufacture some compounds intended for the blood stream.
The bacteria are altered to manufacture some protein in vitro, expanded and tested and then ingested. Theoretically, when we drink milk it can be a signal to the bacteria in the small intestine to make some protein or do some task. Milk or similar drink becomes the trigger that runs our own code, any time we want to run our code we drink a glass of milk, the bacteria detects the milk and runs our code. We can for example engineer a gut bacteria that makes testosterone when we drink some nutrient the bacteria makes the protein when we do not it stops. The proteins can enter the bloodstream through the small intestine.
Antibiotics can be used to eliminate the bacteria and revert to before the experiment.
Open Source White Blood Cell, Exosome, Stem Cell Platform Ex Vivo
A safe blood bacteria would be easier but apparently there are no bacteria or micro-organism colonies living in the blood stream. The only accessible cell is the specific white blood cell living 1 to 7 years in the blood stream.
Another white blood cell called a T-cell is the cell that has been used for providing extra immune response for certain cancers. The cell is programmed to attack cancer cells. We do not want the cell to attack we want the cell to make proteins and inject them into the blood stream. Access to the bloodstream by altering white blood cells in vitro and injecting them into the body. We activate them using some signal, we can add a self-destruct code so that each cell disappears to revert to before the experiment along with antibiotics for the associated bacteria.
A glass of milk activates the gut bacteria to produce a signal molecule into the bloodstream to activate the altered white blood cells to make a protein or a more efficient signal. If I want testosterone I drink milk and so on.
First pass effect. Bio-availability is an issue with some essential proteins so blood stream access is required. The goal might be to add regenerative blood factors that decrease with age.
We also require systemic action, with CRISPR a sequence can be matched before editing. This potentially allows us to search for defects only and do no action if defects are not found, an error checking mechanism on DNA and pass off the DNA as correct. This is important because DNA mutation is thought to be a major problem for every person. Off site effects of CRISPR is a major issue, that needs to be resolved. Mutagenesis.
The only issue is what if the cells trigger an immune response, will we need to differentiate them from real white blood cells. A cytokine storm against itself?
Exosomes are nan particles meaning they are small and can cross the blood brain barrier. These are called engineered exosomes. Exosomes last in the body for up to 8 months. They can be either RNA based, straight to the carbon copy stage which does not use the DNA, or they can be a vesicle to edit DNA called a vector.
AV Vectors Versus Non AV Vectors
The use of a virus to infect the DNA and run some code. This method is a one way deal, it may last some 7 years and has potential safety issues. A virus has DNA or RNA and works by entering cells and injecting its DNA to multiply itself. This behaviour is utilized by replacing the DNA in the virus to a custom DNA so when the virus add its DNA instead its adding the edited DNA. Such virus vectors...
- Retroviruses, Adv. Long-lasting gene expression, efficiently enters cell - DisAdv. Only infects dividing cells, low yield (hard to produce), potential insertional mutagenesis.
- Lentiviruses, Adv. Long-lasting gene expression, will infect dividing and non-dividing cells - DisAdv. Potential insertional mutagenesis.
- Adenoviruses, Adv. Efficiently enters cell, High delivery rate, No chromosomal integration - DisAdv. Immunogenic – rapidly cleared from the body, can cause inflammation and tissue damage,
- Adeno-associated viruses, Adv. Long term expression, wide host cell range - DisAdv. Difficult to produce in high quantities.
- Herpes Simplex Virus, Adv. Produced at high levels, can carry lots of (DNA) - DisAdv. Immunogenic – rapidly cleared from the body, potentially toxic.
- Liposome, Adv. Not immunogenic, can carry lots of (DNA) - DisAdv. Low rate of delivery, transient expression
- Plasmid, Adv. No viral component - DisAdv. Transient expression, difficult to target specific tissues.
- Hybrids. Each have their advantages and disadvantages.
Some emerging approaches are...
- Targeted Cell Delivery, integrative technique of cell mediated transfection where specific antibodies are used to bind DNA to target cells
- Transposons, DNA injected into the bloodstream in lipid capsules. Lipid capsules enter cells. Cells make transposase enzyme from the gene. Transposase cuts out a gene and inserts into a random spot on the genome.
- Antisence technology, antisense drug disrupts translation to result in the prevention of protein synthesis.