Download Recombinant DNA technology

Recombinant DNA technology
Continued
Lecture 3 – 31 March
2015
Prof. Jozef Dulak
Dr Urszula Florczyk
Web: www.biotka.mol.uj.edu.pl/zbm
Milestones of medical biotechnology
Vaccines
a biological preparations that provide active acquired immunity to a particular disease
1797 – Edward Jenner inoculates a child with a viral vaccine to cowpox
(pus from cowpox blisters) to protect him from smallpox.
It took almost 200 years (till 1979) to eradicate smallpox from the world…
Antibiotics
1928 - Penicillin discovered as an antibiotic: Alexander Fleming.
1942 - Penicillin mass-produced in microbes.
1944 - Waksman isolates streptomycin, an effective antibiotic for
tuberculosis.
Edward Jenner's first vaccination
How do vaccines work?
•  Vaccines stimulate the immune system to produce specific antibodies („soldiers”) to fight the
specific virus or bacteria vaccinated for, if it ever attacks the body.
•  The ingredients of the vaccine are eliminated from the body within 24 to 48 hours so no long term
side effects need be worried about.
•  Some vaccines contain live or attenuated viruses/bacteria and are not suitable for immune
compromised individuals.
•  Vaccination programmes preventing polio, measles, mumps, rubella, varicella, rotavirus, hepatitis
and many more have saved millions of lives and are advocated and supported by the World Health
Organisation (WHO).
http://www.everything-i-can.co.za/how-do-vaccines-work.html
Vaccines
Conventional vaccines
1.  Live attenuated organisms (non-pathogenic but immunogenic) – such as the Sabin
oral polyomyleitis, measles and rubella vaccines
2. Inactivated (killed) microorganisms – eg. Salk parenteral poliomyeltis vaccine
3. Subunit vaccines – i.e. one or more antigenic components, as with influenza and
recombinant hepatitis B vaccines
DNA vaccines
4. DNA vaccines – delivery of nucleic acid encoding the pathogenic antigens
RJ Trent – Molecular Medicine, Academic Press 2012
Recombinant DNA technology for vaccines production
Modern subunit vaccines are produced as recombinant proteins
Advantage of higher production yield, incresed
safety, constant and reproducible quality
The encoding DNA sequence is inserted into an
expression vector, most often Escherichia coli, yeast
or a mammalian cell line
The cells are cultured in vitro in bioreactors
Product is purified from the supernatant or directly
from the cells
e.g. Vaccine against human papillomavirus (HPV)…
J. Pongracz, M. Keen – Medical biotechnology, Elsevier 2009
Human papillomavirus (HPV)
•  There are over 100 different types of HPV, and 40 of them affect the genital area
•  For most women and girls the virus goes away on its own or it can develop into cervical cancer,
precancerous lesions, or genital warts, depending on the HPV type.
•  HPV is easy to transmit! HPV is spread mainly by direct skin-to-skin contact
Worldwide estimates on the burden of
HPV & related genital diseases in women
Human Papillomavirus (HPV) is a very common virus
8 out of 10 adults will come into contact with HPV.
Fact
74 % of new HPV infection occur in
people aged 15 to 24, and 80 % of all
women will have the virus at some
point in their lives.
http://www.everything-i-can.co.za/how-many-are-infected.html
Human papillomavirus (HPV) and cervical cancer
- 
- 
- 
There are over 100 different types of HPV, and 40 of them affect the genital area
HPVs 16 and 18 are classified as high risk HPVs causing cervical cancer (about 70%cervical cancer cases)
HPVs types 6 and 11 cause about 90% of genital wart cases
http://www.everything-i-can.co.za/how-many-are-infected.html
Cervical cancer
- 
- 
The second most prevalent cancer among women
Can have several causes – an infection with some type of human papillomavirus (HPV) is the greatest
risk. Unlike other cancers, cervical cancer is not passed down through family genes.
http://www.everything-i-can.co.za/how-many-are-infected.html
How the HPV vaccines are produced?
Produced by recombinant DNA technology, they
are both effective and safe, posing no risk of
infection or cancer
Vaccines against HPV contain viruslike particles
(VLPs)—empty viral capsids with no viral DNA
inside.
http://www.everything-i-can.co.za/how-do-vaccines-work.html
Approved anti-cancer vaccines (1)
Cervical cancer
The U.S. Food and Drug Administration (FDA) has approved two vaccines, Gardasil® and
Cervarix®, that protect against HPV types 16 and 18, that cause ~70% of all cases of cervical
cancer worldwide. In addition, Gardasil protects against infection by HPV types 6 and 11,
which cause an estimated 90% of genital warts cases
Gardasil®, Merck (approved in June 2006); recombinant human papillomavirus
vaccine [types 6, 11, 16, 18]
Cervarix™, GlaxoSmithKline (approved in Australia in May 2007, EU - September 2007)
In December 2014, the FDA approved a nine-valent Gardasil-based vaccine, Gardasil 9, to protect
against infection with the strains covered by the first generation of Gardasil as well as five other
HPV strains responsible for 20% of cervical cancers (HPV-31, HPV-33, HPV-45, HPV-52, and
HPV-58).
Wikipedia
National Cancer Institute
http://www.cancer.gov/cancertopics/causes-prevention/vaccines-fact-sheet
Approved anti-cancer vaccines (2)
Liver cancer
One of the major cause is the hepatitis B virus (HBV) infection
- Two billion people worldwide are infected
- 350 milion become chronic carriers
- Important occupational hazard for health workers
- Approximately 600 000 die annually from complications, such as cirrhosis and
hepatocellular carcionam
- HBV is 50-100 times more infectious than HIV
Anti-hepatitis B virus vaccine – approved in 1981 – first anti-cancer vaccine
- recombinant anti-HBV vaccine released in 1987
Today, most children in the USA are vaccinated against HBV shortly after birth
National Cancer Institute
RJ Trent – Molecular Medicine, Academic Press 2012
http://www.cancer.gov/cancertopics/causes-prevention/vaccines-fact-sheet
Gene replacement, gene knockout, and gene addition
Recombinant DNA technology for understanding the mechanisms
of diseases and development of innovative therapies
Although genetic mapping and comparative sequencing projects can identify associations between
DNA sequence and a disease or human trait , it remains necessary to test this link in relevant model
to develop therapies
-> investigative and translational biomedical research in mouse models
Having identified a target gene,
a strategy must be developed by which the
function of that gene can be repaired
Prerequisites for the creation of a mouse model
o  homologous genes must be identified between human and mouse (more than 98%
genes are shared)
o  measurable biological parameters must be able to be compared between human
and mouse (measurements of heart and lung function, blood pressure in freely
moving animals, magnetic resonance imaging etc)
o  genetic changes in a specific gene in the mouse must result in a phenotype that
correlates with the disease in humans
Genetic modifications in animals
A mouse model have limitations when:
1. homologous genes do not exist between mouse and man
2. the gene defect does not have the same effect in mouse and man
eg. Rb gene, HPRT gene
3. Physiological differences between mice and human:
eg. apoE deficient mice and plaque development – different
than in human
Applied to test:
1. Single gene defects: both inherited and acquired
2. Multigene defects
3. Chromosomal defects
Transgenic animals
Transgenic animals are genetically modified organisms (GMOs) which their genetic
material changed
1.  Investigation of the mechanisms of disease
2.  Testing new therapies
3.  Producing new drug
First transgenic mice
A DNA fragment containing the promoter of the mouse
metallothionein-I gene fused to the structural gene of rat
growth hormone was microinjected into the pronuclei of
fertilized mouse eggs.
Dramatic growth of mice that develop from eggs microinjected
with metallothionein-growth hormone fusion genes.
implications for studying the biological effects of growth
hormone, as a way to accelerate animal growth, as a model for
gigantism, as a means of correcting genetic disease, and as a
method of farming valuable gene products
Biochemistry. 5th edition.
Berg JM, Tymoczko JL, Stryer L.
New York: W H Freeman; 2002.
Palmiter et al., Nature 1982: 300: 611-615
Two methods of producing transgenic mice
Transgenic mice can be obtained by two major ways:
-  transfer of cloned DNA into fertilized oocytes and cells from very early stage embryos
-  transferring the transgene into cultured embryonic stem cells.
Transgene can be inserted into
mouse genome either randomly or it
may be incorporated in the chosen
locus inside the mouse genome. The
second strategy is called targeted
approach and it is based on the
homologous recombination.
http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/T/TransgenicAnimals.html
Strategies for creation of mouse models of human diseases
1. Disease-driven, directed genetics – a human mutation is identified and then
a specific mouse model is made to mimic it
Type of genetic modifications:
a. transgenesis – integration of DNA sequences randomly into the genome
b. gene targeting – precise modification by homologous recombination–
gene is introduced into its normal genomic location: „knock-out”, „knock-in”
c. chromosome engineering
2. Mutagenesis-driven, non-directed genetics – relies on the
selection of disease phenotypes following random mutagenesis,
induced by chemicals or gene trapping
eg. Mdx mice (Duchenne muscular dystrophy_
db/db mice – mice lacking leptin receptor
Transgenesis – integration of DNA sequences randomly into
the genome
-  heritable integration
Method:
pronuclear injection of fertilized mouse oocytes with the linearized DNA transgene
Large numbers of oocytes are
produced from immature female
mice by administering hormones
which lead to superovulation
The superovulated female is
then mated and one-cell stage
embryos are collected.
one-cell stage embryos are
injected with the transgenic
DNA into the large pronucleus
- - random integration can lead to inadequate expression pattern
J Pongracz, M. Keen - Medical Biotechnology, 2009
Conventional transgenes
•  A promoter
Jazwa et al. Gene 2013
•  An intron element to mimic normal gene
structure
•  The coding sequence of the molecule
under investigation
•  A polyadenylation signal
J Pongracz, M. Keen - Medical Biotechnology, 2009
An alternative method for introducing foreign genes or
targeted gene sequences into mice is to use
murine embryonic stem (ES) cells
What are embryonic stem cells?
pluripotent stem cells derived from the inner cell mass of a blastocyst,
an early-stage preimplantation embryo
- develop from eggs fertilized in vitro
-  derived from 4-5 days old embryos
- isolated from ~ 8 cell embryo of inner cell mass
Kaur et al., Journal of Diabetology, 2012
Murine embryonic stem cells
Wikipedia
Sir Martin Evans, Mario Capecchi, Olivier Smithies
for their discoveries of principles for introducing specific gene modifications in mice by the
use of embryonic stem cells
Nobel Prize 2007
The transgenic mouse generation by the method with
the use of embryonic stem cells
The transgenic mouse generation by the method with the use of embryonic
stem cells involves:
-  the ES cell cultivation in vitro
-  DNA introduction usually by electroporation
-  positive clone selection due to the presence of the selectable marker gene
in the introduced DNA
Difficulties can be connected with the random transgene integration event
and with the germline transmission of the transgenic ES cells
The method is commonly used for the DNA integration by homologous
recombination in order to obtain knockouts and knockins
– GENE TARGETING
Gene targeting - precise modification
-  enables the removal or replacement of specific gene by homologous recombination
-  key – EMBRYONIC STEM CELLS (ES cells)
genetically modified ES cells
Method:
- once reimplanted into the uterus of a pseudopregnant foster mother, the injected „donor” ES
cells compete with cells in the host blastocyst
to form the developing embryo and ultimately
lead to a chimeric mouse
-  if the germ cells of the chimera also contain
cells derived from the donor ES cells, some
progeny resulting from mating will have one
set of chromosomes derived completely from
the donor , thereby establishing a „line”
J Pongracz, M. Keen - Medical Biotechnology, 2009
Gene targeting – gene knock-out
Most widely applied use of homologous recombination in ES cells has been the generation of
knockout mice
- Usually all or part of the protein coding sequences is deleted or an insertion is made into a
exon encoding an essential domain
- - the targeting vector is introduced into ES
cells by subjecting cell suspension in a
solution of DNA to a short electric pulse
- - ES cells that succesfully exchange the
targeting sequences by homologous
recombination for one of its two
corresponding genomic sequences are able
to grow in the presence of the selective
antibiotic
- ES clone is then injected into blastocysts to
ptoduce a chimera
J Pongracz, M. Keen - Medical Biotechnology, 2009
Gene knockout technology
Molecular biology of the gene
Knockout of VEGF is lethal in heterogenous form
Ferrara & Allitalo, Nature medicine, 2000
Gene targeting – conditional gene deletion
Use of cre recombinase for conditional knockouts
Most widely used: Cre recombinase and its
32 base recognition element, loxP
A gene is engineered by homologous
recombination in ES cells so that the whole
gene or an exon encoding crucial protein
domain , is flanked by recognition sites for
a recombinase enzyme that can delete the
intervening sequences
Gene knockout is restricted
by expressing the recombinase
in specific tissueas or at particular time
Molecular biology of the gene
Gene targeting - knock-in
- The majority of diseases with genetic basis involve small sequence changes
- The same ES-based approaches can be used to create mouse lines to model such
mutations – so called „knock-in”
Small mutational changes (asterisk) can be introduced by targeting and usually
it is desirable to remove the selection cassette by recombinase system
J Pongracz, M. Keen - Medical Biotechnology, 2009
Jazwa et al. Gene 2013
Apolipoprotein E
A multifunctional glycosylated protein, mostly involved in the transports of
lipoproteins and cholesterol
Human apoE is a 299 amino acid protein that occurs in three major isoforms (apoE2,
apoE3 and apoE4) encoded by three APOE alleles (ε2, ε3 and ε4) differing with
respect to the presence of cysteine or arginine at two polymorphic sites.
LDLR
binding
•  ApoE3, the most
common isoform,
has cysteine at
amino acid
position 112 and
arginine at 158
•  ApoE2 has cysteine
at both 112 and 158
•  ApoE4 has
arginine at both
sites
Lipids
binding
Polymorphism of apolipoprotein E
ApoE exerts its biological functions, especially lipid transportation, by
binding to its receptors, the low-density lipoprotein receptor (LDLR) family.
There is only one isoform of apoE in rodents.
Murine apoE preferably associates with HDL, and its
clearance is mainly through LDLR
In human:
-  The most common allele is e3, which is found in more than half of the general
population.
-  ApoE4 preferentially binds to lower density lipoproteins and is associated with
increased risk of atherosclerosis and neurodegenerative disorders, including
Alzheimer Disease
-  The APOE e2 allele has been shown to greatly increase the risk of a rare condition
called hyperlipoproteinemia type III.
Apolipoprotein E knockout mouse
The Apolipoprotein E knockout mouse model is one of the most widely used experimental model of
atherosclerosis. These mice rapidly develop atherosclerotic lesions that resemble human lesions evolving
over time from initial fatty streaks to complex lesions.
PATHOPHYSIOLOGICAL FEATURES
Cardiovascular features:
• Apolipoprotein E deficiency directly results in the increase of plasma levels of LDL and VLDL.
•  Very high cholesterol level (500-600 mg/dl instead of 50-100 mg/dl)
• Spontaneous development of atherosclerotic lesions throughout the arterial tree appearing first in the
aortic arch in young mice and progressing in the thoracic and abdominal aorta in older mice
Maeda et al., Atherosclerosis 2007
En face preparations of oil red O stained aortas from C57BL6/J and
ApoE KO mice at 26 weeks of age (from Behr-Roussel et. Al, 2006).
Polymorphism of apoE – knock-in technology
To assess allele-specific differences in apoE function in vivo, it is desirable to
have mouse strains in which the murine apoE is replaced by human apoE
isoforms expressed under the natural regulation of this protein.
KNOCK-IN technology
•  Homozygous for a human APOE4 (or APOE3 or APOE2) gene targeted
replacement of the endogenous mouse Apoe gene
•  Expresses human apoliprotein E4 isoform under the control of the murine
Apoe regulatory sequences
RIKEN BioResource Center (BRC)
Generation of human apoE4 knock-in mice
– similar strategy for ApoE3 and ApoE2
Targeting strategy for apoE4 knock-in mice and homologous intergration of the transgene. (A)
Schematic diagram of the knock-in targeting strategy. (Top) The structure of the endogenous Apoe locus
including exons 1–4 (black boxes). (Middle) The targeting vector containing the human apoE4 cDNA (hu
cDNA). (Bottom) The predicted structure of the knock-in allele after homologous recombination. The
neomycin-resistance (neo) and thymidine kinase (TK) genes are for selection of the targeted ES cells. The
neo cassette is flanked by 34 bp loxP sequences (triangles). pA represents the endogenous polyadenylation
signals. Restriction sites: B, BglII; E, EcoRI; H, HindIII; N, NcoI; S, SalI; X, XmnI. (B) Southern blot analysis of
tail-tip DNA from wild-type (+/+), heterozygous (4/+) and homozygous (4/4) knock-in mice digested with
HindIII and hybridized with the 3′ probe shown in (A). The wild-type Apoe allele generates an 8.0 kb HindIII
fragment, whereas the targeted allele yields the diagnostic 6.4 kb HindIII fragment.
Hamanaka et al., Hum Mol Genet. 2000; 9(3): 353-361
Strategies for creation of mouse models of human diseases
1. Disease-driven, directed genetics – a human mutation is identified and then
a specific mouse model is made to mimic it
Type of genetic modifications:
a. transgenesis – integration of DNA sequences randomly into
the genome
b. gene targeting – precise modification – gene is introduced
into its normal genomic location: „knock-out”, „knock-in”
c. chromosome engineering
2. Mutagenesis-driven, non-directed genetics – relies on the
selection of disease phenotypes following random mutagenesis,
induced by chemicals or gene trapping
eg. Mdx mice (Duchenne muscular dystrophy_
db/db mice – mice lacking leptin receptor
Chromosome engineering
• 
Aberrations in human chromosome copy number and structure are
common and deletorious, both as development-associated defects and as
aqcuired events in tumors (e.g. trisomy of chromosome 21 in Down
syndrome)
• 
The combination of gene targeting in ES cells, recombinase technology and
other techniques makes it possible to generate new chromosomes carrying
specific and defined deletions, duplications, inversions and translocations,
which can serve as models for human chromosomal aberrations
Cre-loxP technology can be used:
- To create very large deletions or inversions on a single chromosome
-  To bring about specific recombination between two different
chromosomes
SummaryStrategies for generating knockout/transgenic mice
The Metabolic and Molecular Basis of Inherited Diseases, McGraw Hill, 2001
Strategies for creation of mouse models of human diseases
1. Disease-driven, directed genetics – a human mutation is identified and then
a specific mouse model is made to mimic it
Type of genetic modifications:
a. transgenesis – integration of DNA sequences randomly into
the genome
b. gene targeting – precise modification – gene is introduced
into its normal genomic location
c. chromosome engineering
2. Mutagenesis-driven, non-directed genetics – relies on the
selection of disease phenotypes following random mutagenesis,
induced by chemicals or gene trapping
eg. Mdx mice (Duchenne muscular dystrophy_
db/db mice – mice lacking leptin receptor
Mutagenesis-driven, non-directed genetics
This approach requires selection of phenotypes following either spontaneous or
induced random mutation
Random mutagenesis strategies can be divided based on mutagen used:
- Chemical mutagenesis
Alkylating mutagen (N-ethyl-N-nitrosourea)
-  ENU-treated males are mated with females, progeny are put into a breeding
programme
-  Phenotypic screening
- Gene trapping using transgene insertion
as the mutagen
Gene trapping using transgene insertion
Reporter gene is activated following insertion into an endogenous transcription unit
-  Gene trap vectors contain a splice acceptor sequence upstream of a reporter and are
activated following insertions into introns
- - prevents expression of gene sequence downstream of the site of insertion
Based on the pattern of reporter expression in vitro, ES cell clones are tested in vivo:
o  For expression of the reporter in chimeric embryos following introduction into
blastocysts
o  then, for both reporter expression and function of the trapped gene in embryos and
adults, if germline transmission can be achieved
Drug testing using mouse models
DRUG DISCOVERY
e.g. If a drug candidate is thought to operate through a particular gene product
this can be tested by comparing the effect of the compound in wild type and
mice mutated at the relevant gene locus
Also to study in vivo efficacy of candidate compounds – mice can be humanized with
respect to small subgroups of proteins responsible for the major routes of transport and
metabolism of a large proportion of drugs
Heme oxygenase-1
PRODUCT
MECHANISM
BILIVERDIN
BVR
ROS scavenging
Inhibition of complement
antioxidant
anti-inflammatory
anti-apoptotic
pro-angiogenic
cytoprotection
ferritin
synthesis
iron ATP-ase
pump
antioxidant
anti-inflammatory
anti-apoptotic
cytoprotection
BILIRUBIN
HEME
HO-1
Fe2+
activation of sGC leading
to cGMP production
CO
ACTIVITY
p38 MAPK regulation
others
anti-apoptotic
anti-proliferative
anti-thrombotic
antiinflammatory
pro-angiogenic
cytoprotection
Dulak et al. Circulation 2008
Investigation on the role of heme oxygenase-1
in human diseases
1.  Inducers of heme oxygenase-1 expression/activity
2. Inhibitors of Hmox-1 activity
A) small molecular compounds
b) siRNA
3. Heme oxygenase-1 knockout mice
4. Heme oxygenase-1 transgenic mice
Wound healing is dependent on HO-1
Inhibition of HO-1 attenuates wound healing
PBS
SnPP s.c.
Lack of HO-1 attenuates wound healing
SnPP i.p.
Day 0
Day 3
Day 7
Lack of HO-1
impairs blood vessel
formation in wounds
Grochot-Przeczek et al. , PLoS ONE 4(6): e5803; 2009
Deshane et al., J Exp Med., 2007,
Development of stable transgenic mouse line
overexpressing HO-1 in the skin
To study skin related processes,such as wound healing, cancerogenesis, psoriasis…
ES cell- based, random DNA insertion strategy
ES cell cultivation in vitro, DNA introduction by
electroporation and positive clone selection due to the
presence of the selectable marker gene in the
introduced DNA.
After the amplification and confirmation of the
transgene presence, ES clones are microinjected into
mouse blastocyst.
The founders derived from the embryos that have
undergone the transgene integration will be chimeric,
composed of a mixture of transgenic ES cell and wild
type host cells. If the transgenic ES cells have
contributed to the germ line of chimeric mice, their
offspring can be fully heterozygous and after crossing
these heterozygotes to each other, can develop
homozygous mice strain.
Modified from:
Human Molecular
Genetics 2 2nd ed.
New York and
London: Garland
Science; c1999.
From ES cells to transgenic mice:
microinjection of ESC into the murine blastocyst
Random insertion into the genome results in an indiscriminate transgene incorporation into chromosomes
and that strategy is commonly used for generation of transgenic mice which overexpress the gene of interest.
3 lines of transgenic mice:
•  K14HO-1 clone 9
•  K14 HO-1 clone 78
•  K14 HO-1 clone 94
Germline
transmission
Genotyping by PCR and Southern blotting
PCR with primers
specific for human HO-1
Southern blotting with probe specific for human HO-1
Higher expression of HO-1
in epidermis of KER14-HO1 mice
Skin
HO-1+/+
Tg
HO-1Tg
WT
Higher expression of HO-1
in primary murine keratinocytes of KER14-HO1 mice
Primary murine keratinocytes culture:
2 days after isolation
Tg
Wt Tg Wt
Tg
Grochot-Przeczek et al. , PLoS ONE 4(6): e5803; 2009
HO-1 is required for blood vessel formation
Inhibition of HO-1 attenuates wound healing,
while its overexpression in the skin promotes it
100
Wound closure [% of day 0]
Skin wound healing
90
HO-1-KO
HO-1-HT
80
70
60
HO-1 WT
HO-1 Tg
50
40
30
20
10
0
0
1
2
3
4
5
6
7
8
Day after wounding
10
11
12
13
Grochot-Przeczek et al. , PLoS ONE 4(6): e5803; 2009
Transgenic and knockout mice are indispensable tools for
investigation the mechanisms of diseases and effectiveness
of therapies
Transgenic animals for drug production
Orphan diseases and recombinant DNA technology
Orphan disease - a disease that has not been „adopted” by the pharmaceutical industry
because it provides little financial incentive for the private sector to make and market
new medications to treat or prevent it.
An orphan disease may be a rare disease (according to US criteria, a disease that affects
fewer than 200,000 people) or a common disease that has been ignored (such as
tuberculosis, cholera, typhoid, and malaria) because it is far more prevalent in
developing countries than in the developed world.
Transgenic animals as drug factories
An anticoagulant, human anti-thrombin - a natural serum
protein with anti-thrombotic and anti-inflammatory
properties.
used for the prevention of blood clots in patients with a rare
disease known as hereditary antithrombin (AT) deficiency;
from the milk of goats
It is used in obstetrics, treatment of deep vein thrombosis
• ATryn – first drug produced by transgenic goats, which has been registered by the European
Commission in July 2006
•  The FDA approval (2009) of the first biological product produced by genetically engineered
animals recombinant human antithrombin.
Because hereditary AT deficiency occurs in a small population (approximately 1 in 5,000 people in
the United States), the FDA granted ATryn an orphan drug designation.
One GM goat can produce the same amount of antithrombin in a year as 90,000 blood donations
Atryn – first approval for a biological product produced by genetically
engineered animal
http://pharmacologycorner.com/fda-approves-atryn-recombinant-human-antithrombin-for-the-treatment-ofhereditary-antithrombin-deficiency/
Biotechnology - achievements
Biotechnology therapeutics approved by the U.S. Food and Drug Administration
(FDA) to date are used to treat many diseases, including leukemia and other cancers,
anemia, cystic fibrosis, growth deficiency, rheumatoid arthritis, hemophilia, hepatitis,
genital warts, and transplant rejection.
Biotechnology has created:
* more than 200 new therapies and vaccines, including products to treat
cancer, diabetes, AIDS and autoimmune disorders.
* more than 400 drug products and vaccines currently in clinical trials
targeting more than 200 diseases, including various cancers, Alzheimer’s
disease, heart disease, diabetes, multiple sclerosis, AIDS and arthritis.
* hundreds of medical diagnostic tests for early detection of diseases, for
keeping the blood supply safe, or for detection of pregnancy at home.
* DNA fingerprinting, which has dramatically improved criminal
investigation and forensic medicine.
Next lecture
Gene therapy
14 April, 2015