Genetic Technology and Medicine

- Human proteins, other than insulin, produced by genetic technology include human growth
hormone, thyroid stimulating hormone, and factor eight (clotting protein)
- Advantages of using bacteria, yeast and cultures of mammalian cells to produce human
proteins:
Simple nutritional requirements
Large volumes can be produced
Production facilities do not require much space
The processes can be carried out almost anywhere in the world
There are few practical and ethical problems because proteins do not have to be extracted
from animals or collected from many donors
There is no risk of infection from HIV due to donated blood.
- Disadvantages:
Bacteria do not modify their proteins in the same way that eukaryotes do. Thus, it is much
better to use eukaryotic cells to produce human proteins.
- Genetically modified hamster cells have been made to produce factor eight.
The gene coding for factor eight is inserted into hamster kidney and ovary cell, which are then
cultured in fermenters. The cells constantly produce factor eight which is extracted and
purified before being used to treat people.
- Adenosine deaminase (ADA), an enzyme used to treat sever combined immunodeficiency
disease (SCID), are made by a genetically modified larva, the cabbage looper moth caterpillar.
This enzyme is administered to patients while they are waiting for gene therapy or when
therapy is not possible,
- Sheep (alpha-antitrypsin for emphysema) and goats (antithrombin to stop blood clotting) have
been genetically modified to make human proteins in their milk.

Genetic Screening
- An example of a disease that could be screened for is breast cancer (faulty Brca-1 and Brca-2
alleles)
- Advantages:
Helps to provide early diagnosis
Genetic tests can identify that embryos have a genetic condition.
Termination can be done.
Counselling can be offered
Couples who are tested and who are both carriers can use the information to make decisions
about starting a family.
It allows people to prepare for late onset Huntingtons Disease.
People can take preventative measures against the disease such as getting an elective
mastectomy if they are positive for the Brca-1, -2 alleles.
- Disadvantages:
Amniocentesis can increase the risk of miscarriage.
Psychological problems may arise due to the stress of knowing about having a disease.
It is expensive.
Couples who are both carriers may divorce or not get married.
Sex pre-selection can occur.
- Pre-implantation genetic diagnosis (PGD) is a technique involving the mixing the fathers sperm
with the mothers eggs (same as normal IVF), then at the eight-cell stage, removing one of the
cells. The DNA in the cell is analysed and used to predict whether or not the embryo will have a
genetic disease for which both parents are carriers. An embryo not carrying the allele that would
cause the genetic disease is chosen for implantation and the others are discarded.
This has been used to avoid diseases such as Duchenne muscular dystrophy, thalassaemia,
haemophilia, and Huntingtons disease.
- Thalassaemia is a blood disease similar to sickle cell anaemia, common around the
Mediterranean. Due to genetic screening advice could be given tot couples with the disease
and already pregnant women could receive advice about termination. This has lead to a
decrease in the incidence of thalassaemia.
- Huntingtons disease is a late-onset disease with no cure. Very rarely, the dominant allele
for Huntingtons is not even expressed. But, getting screened leads to many ethical
dilemmas involving factors such as mental health and family planning.
- Termination for a medical reason, rather than for any other, is called a therapeutic abortion.
 This has also been used to select an embryo that did not have a disease and did have a
tissue type that would allow a successful transplant into a sick elder sibling.
- Amniocentesis and chorionic villus sampling is used to screen a fetus while in the uterus.
Amniocentesis is used to obtain a sample of amniotic fluid at 15 to 16 weeks of pregnancy.
They are usually used to look for chromosomal mutations (e.g. Downs syndrome).
- Ultrasounds scanning is used to visualise the fetus and locate the position of the placenta,
fetus and umbilical cord, A suitable point of insertion for the hypodermic syringe needle is
found and a mark is made on the spot. This position is away from the fetus, umbilical cord
and placenta.
Chorionic villus sampling can be carried out between 10 and 13 weeks of pregnancy.
- It allows an earlier warning of genetic problems than Amniocentesis.
- A small part of the placenta called the amnion is removed with a thin needle. This process is
monitored via ultrasound.
- It increases the miscarriage rate by 1-2%. The original miscarriage rate is about 2-3%. The
procedure is riskier with chorionic villus sampling than with amniocentesis after 15 weeks.

Gene Therapy
- Vectors for genetic therapy:
Liposome
Retrovirus
Lentivirus: Though they insert their genes into the hosts DNA, they can be modified to
inactivate replication. HIV has been inactivated to act the same way.
Adeno-associated virus (AAV): It does not insert its genes into the host genome, so its genes
cannot be passed to daughter cells by cell division. This is problematic for short-lived cells.
- Naked DNA: Occasionally, DNA is inserted into tissues without any vectors. It is used in trails for
gene therapy for skin, muscular, and heart disorders. It has the advantage of not having the
problems associated with using vectors.
- Examples of gene therapy:
Treating SCID:
 - SCID is and immunodifeciency disease. It causes sufferers to die in infancy or have to live
in a bubble. It is caused by the inability to make adenosine deaminase (ADA).
- Some T lymphocytes were removed from the patient and the normal alleles of the ADA
gene were introduced using a virus as a vector. This was not a permanent cure because the
T lymphocytes would break down and not divide, so the treatment would have to be
repeated.
- Stem cells harvested from bone marrow were also used in gene therapy, but this was
stopped because many patients got leukaemia from using a retrovirus as a vector.
Retroviruses insert their genes randomly into the hosts DNA. This can cause the genes
to be inserted in the middle of another gene or a regulatory sequence of a gene, causing
cancer.
Leber congenital amaurosis, a form of hereditary blindness in which retinal cells die off from
an early age.
The normal beta-globin allele was inserted into the blood stem cells of patients in order to
correct beta-thalassaemia.
Haemophilia B
Cystic fibrosis

Cystic Fibrosis (CF) as an Example

- Symptoms:
Abnormally thick mucus is produced in the lungs and other parts of the body.
High susceptibility to bacterial infections in the lungs because it is difficult to remove the
mucus, allowing bacteria to breed in it.
The pancreatic duct may be blocked.
Men with CF often are sterile because thick secretions block the ducts of their reproductive
systems.
- Simple treatment:
Percussion therapy: Helps patients cough up mucus by pummelling against the back to loosen
the thick mucus.
Taking pancreatic enzymes by mouth.
- Cause:
It is caused by a recessive allele of the gene that codes for the transporter (CFTR) protein.
CFTR is found in the lungs and other parts of the body and allows chloride ions to leave cells.
The recessive allele codes for a protein that does not act as a chloride ion transporter.
Usually, the CFTR proteins lining the lungs pump chloride ions out of the cells, creating a high
concentration of chloride ions outside the cells. This lowers the water potential outside the
cells, causing water to move down a water potential gradient by osmosis out of the cell and
into the mucus. The mucus, thus, becomes thin enough to be removed by the beating of cilia.
Without the CFTR proteins, the mucus remains sticky and hard to remove.
- The CFTR gene and gene therapy:
The CFTR gene is found on chromosome 7.
 - The most common mutation is the deletion of three bases. One amino acid is missing and
the cell machinery recognises this and does not place the CFTR protein into the cell surface
membrane.
- Another kind of CF is caused by a base substitution which codes for a stop codon in the
middle of the gene. When the gene is transcribed, only a short length of CFTR is made.
A carrier makes enough of the normal protein to remain healthy.
Because CF is determined by a single gene and a recessive one at that, it is a good candidate
for gene therapy.
- If one normal dominant allele is added, the correct CFTR should theoretically be made.
There have been several attempts to treat CF with genetic technology:
- The normal allele was inserted into liposomes which were then sprayed as an aerosol into
the noses of nine volunteers. This introduced the allele into the cells in the lining of the
nose, but since those cells are short-lived, this was not very efficient.
- A normally harmless virus was also used as a vector. The alleles were introduced in some
gas-exchange cells, but the trial was stopped after some patients experienced unpleasant
side-effects from the infection.
- A drug, administered orally, called PTC124, is being used to allow translation to just keep
going across the entire codon in the case of the substitution mutation, so that the entire
protein is made.

Somatic and Germ Cell Gene Therapy

- Somatic cells are body cells.
- Germ cells are cells involved unisexual reproduction such s gametes and embryos.
- In theory, a woman with CF could have IVF, but insert the normal allele for the CFTR gene into
the egg before it is fertilised by sperm.
This is illegal now because all the cells in the child would be from the genetically engineered
zygote, so they would be able to pass down this allele to their children. When this happens the
allele is said to be germ line, being passed from generation to generation.
 Also, there is a high have of offspring produced in such a way of having other unpredicted
diseases.