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There are several ways in which gene therapy is carried out. One way is to correct base substitutions such as Sickle Cell Anemia. In this method the erroneous base pair is corrected as follows


· A short string of nucleotides is made called an OLIGOMER. This oligomer is exactly complementary to the section of the gene where the error is located.the only difference being that the oligomer has the right combination as found on a normal gene while the erroneous one has the error.


· The oligomer then binds to its complementary sequence on the DNA molecule, and causes a bulge due to the error.


· The cell where this erroneous gene is found has special DNA repair mechanisms that detect the bulge.


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· The mechanism causes enzymes to be sent to the erroneous site, and there they substitute the erroneous base, by complementing the oligomer that has the complementary sequence for the correct base.


Below are some diagrams and a brief explanation





Gene correction exploits cellular proofreading enzymes that detect errors in DNA and make corrections. Scientists place into the cell a small hybrid RNA-DNA molecule called a chimeric oligomer that pairs with the defective gene in the region of the error (a). Repair enzymes use the oligomer as a template to guide the correction. Seen close up (b), the oligomer binds snugly with the defective gene except in the region of the error, where the mismatch causes a bulge. Repair enzymes detect this bulge and replace the erroneous nucleotides. In this example the guanine (G)-cytosine (C) pair is incorrect. The oligomer provides the template indicating that an adenine (A)-thy mine (T) pair should be inserted in that spot. The repair enzymes follow the instructions in the template and correct the gene accordingly. Corrections made this way endure for generations of cell divisions.


The other form of gene therapy involves the following method, using the example of gene therapy for familial hypercholesterolemia


· There is a virus that is used as a vector. The harmful genes of the virus are thoroughly removed leaving only the genes that allow the viruses DNA to combine with the host’s( in this case the human) chromosomes. They then insert the corrective gene that is erroneous in the patients cell, that is causing the patient to suffer from the disorder. This will allow the vector to insert the corrective gene by splicing the chromosome and inserting this gene, and then replacing the erroneous one.


· Special prompters in the form of chemicals activate the corrective genes to allow them to start their activity in the body. These are timed and ensure that the prompters work only when in the body and not outside. The retroviral also enable the organs which can benefit from this type of treatment to be diverse.


· Then a piece of the liver is removed and then cultured in a petri dish. The virus is then prepared as seen above, and the corrective gene in this case will be the LDL-receptor gene, that is essential to allow the liver to accept and use LDL cholesterol that is good for the body. Once the virus contains this gene, it is introduced into the culture of the liver cells. The virus then move to the receptors of the cell where they begin to work their way into the cell. Once inside the virus releases the gene, and the gene moves towards the nucleus and to the chromosome containing the erroneous gene. The virus splices the erroneous gene and replaces it with the LDL receptor gene that is the correct gene. Splicing of the gene is characteristic of all viruses. The cells in the culture that refuse to accept the retroviral are removed from the culture. The repaired cells are then infused back onto the liver through the portal vein.


· The viruses have specific receptors which they combine with and these restrict the cells which the virus may have an effect on


· Examples of diseases treated in this way include sickle cell anemia, muscle dystrophy, cystic fibrosis, and most recently some forms of cancer. All these examples apply the same basic principles as above for gene therapy.





Here the virus is being converted into a vector so as to enable it to be used in gene therapy.





This series of diagrams shows how the virus replaces the erroneous DNA and corrects the gene. This is explained further in the last three bullets above.





The table shows the various advantages of using various forms of viruses and non-viral vectors.





The table shows the various approved disorders that gene therapy has been used for.





the diagram summarizes the process of gene therapy for a patient suffering from familial hypocholesteroloemia.





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