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The therapeutic gene to be transferred is extracted from the cell of a healthy individual. The gene is extracted by cutting the DNA using a restriction enzyme (restriction enzymes "digest" DNA at designated nucleotide locations along the DNA chain). There are different types of restriction enzymes, each being specific to the location of the DNA chain that it will cut. The section of cut DNA has to be intact then a technique called electrophoresis is used to separate the selected pieces of DNA and remove the genes that contained the DNA sequence that coded for the polypeptides needed.
Similar restriction enzymes are used to remove a section of enzyme from a plasmid or virus and using ligase (a special enzyme used to "glue" a foreign piece of DNA into a donor cell) the therapeutic DNA sequence is placed in the DNA of the vector. In the case of a virus, instead of causing illness the virus carries the normal genes into the target cells where they begin functioning.
Ex Vivo
There are methods of inserting genes, such as addition of chemicals to allow the large DNA molecules in solution to pass through the cell membrane. This requires that the cells be removed from the body and treated in a laboratory, making it suitable for treatment of easily transplantable tissues such as blood and bone marrow. This type of gene therapy is referred to as ex vivo, as the cell to be implanted are harvested from the patient, modified and then replaced.
In Vivo
A technique that involves direct insertion of the vector into the patient is referred to as in vivo gene therapy. There are also other techniques, which include laser micropuncture of the cell membrane, electroporation and biolistics (using high velocity tungsten microprojectiles to insert DNA)
Activity
Pair up On a sheet of paper, list down at least 3 ethical concerns connected with HGP and the Germ line therapy
Questions
We can now identify who has what type of sickness, what kind of ethical concerns (in finding a job, family, etc.), are connected to this? And given these concerns should we still pursue the HGP?
Questions
There is a way for us to live without any sickness, prolong our lives and manipulate the way our children will come out. Again, what sort of issues may arise and given those issues, should we still pursue HGP and Gene Therapy?
References
http://www.ndsu.edu/pubweb/~mcclean/plsc431/students99/rothe.htm
http://sgugenetics.pbworks.com/w/page/38231663/The%20Human%20Genome%20Project
http://www.montegen.com/Montegen/Nature_of_Business/The_Library/Genomics/Germ_Li ne_Gene_Therapy/germ_line_gene_therapy.htm