History of Genetic Engineering Tools of Genetic Engineering Principles of rDNA technology Applications of Genetic Engineering in agriculture medicine and orthodontics

1. GENETIC ENGINEERING
Presented by
Ashok kumar A
Dept of orthodontics

2. CONTENTS
• Introduction
• History of Genetic Engineering
• Tools of Genetic Engineering
• Principles of rDNA technology
• Applications of Genetic Engineering
• Disadvantage
• Conclusion

3. GENETIC ENGINEERING
• This is a common occurrence within the same species.
• But by artificial means, when a gene of one species in
transferred to another living organism, it is called
recombinant DNA technology.
Genetic recombination is the exchange of information
between two DNA segments.

4. • This refers to the processes that allow introduction of new
DNA into a cell.
• This allows the taking of genes from one organism and
placing them in the chromosomes of another producing a
transgenic organism.
The genes can come from individuals of the same species
(eg transferring a healthy gene from one person into a person with a
genetic disease = gene therapy) OR
From a different species (eg placing insulin producing gene into a
bacteria)

5. History of Genetic Engineering
• In 1951-The term "genetic
engineering" was first coined
by Jack Williamson in his
science fiction novel
(Dragon's Island).
• In 1953, James Watson and
Francis Crick proposed the
double helix structure of
DNA.

6. • 1972- Paul berg created the first recombinant DNA molecules by
combining DNA from the monkey virus SV40 (Simian virus 40)
with that of the lambda virus (bacteriophage)
• 1973- The first GMO was a bacterium generated by Herbert
Boyer and Stanley Cohen

7. • In 1980s invention of polymerase
chain reaction (PCR).
• Development of genetic
fingerprinting.
• Transgenic mice produced
carrying human genes.
• Transgenic plants produced
resistant to a herbicide.
• In1996 The birth of the first
cloned animal, Dolly the sheep,
was announced.

8. Tools of Genetic Engineering
Restriction Endonucleases
• Bacterial enzymes that can cut/split DNA at specific sites. These
were first discovered in E.coli restricting the replication of
bacteriophages ,by cutting the viral DNA.
Recognition sequences:
• Recognition sequence is the site where the DNA is cut by a
restriction endonuclease.
• Restriction endonucleases can specifically recognise DNA with a
particular sequence of 4-8 nucleotides & cleave

9. Cleavage patterns:
• The cut DNA fragments by restriction endonucleases may have mostly
sticky ends or blunt ends.
• DNA fragments with sticky ends are particularly useful for rDNA
experiments,since single stranded sticky DNA ends can easily pair with
any other DNA fragment having complementary .

10. DNA ligases
• These were originally isolated from viruses, also occur in E.coli
& eukaryotic cells.
• The cut DNA fragments are covalently joined together by DNA
ligases

11. Host cells
• The hosts are the living
systems or cells in which the
carrier of rDNA molecule or
vector can be propagated.
• Microorganisms are preferred
as host cells, since they
multiply faster compared to
cells of higher organisms.

12. E.coli
• This was the first organism used in the DNA technology experiments.
• The major drawback is that it cannot perform post translational
modifications.
Eukaryotic Hosts
• These are preferred to produce human proteins, since these have complex
structure suitable to synthezise complex proteins.
• Mammalian cells possess the machinery to modify the protein to the
active form.(post translational modifications)
E.g., Tissue plasminogen activator

13. Vectors
• DNA molecules, which can carry a foreign DNA fragment to be
cloned.
• These are self replicating in an appropriate host cell.
- Plasmids - E.coli
- Bacteriophages - phage λ, phageM13
- Cosmids
- Artificial chromosome vectors.

14. Principles of rDNA technology

17. GENE THERAPY - TYPES
SOMATIC:
The somatic gene therapy involves changes in the target cells,
however, that are not transferred to the next generation.
GERM LINE THERAPY:
Germ line gene therapy, germ cells i.e. sperm or eggs are
modified by the introduction of functional genes, which are ordinarily
integrated into their genomes. Therefore, the change due to therapy
would be heritable and would be passed on to later generations.

20. APPLICATION OF GENETIC
ENGINEERING

21. AGRICULTURE

22. Genetically modified organism

23. Applications of Genetic Engineering in Medicine
• Human insulin gene inserted into
the bacterium E.coli to produce
synthetic "human" insulin, for the
treatment of insulin-dependent
diabetes .
• Today, the microorganism (such
as: yeast) is used to produce virus
antigen used as a vaccine, that
stimulate human immune system
against this virus.

24. • Producing human growth hormones:
to treat growth retardation (dwarfism).
• Producing Follistim injection:
(contains the FSH hormone) for
treating infertility.
• Other biopharmaceuticals under
development through genetic
engineering, include: anti-cancer drug
and a possible vaccine for AIDS,
malaria, etc

26. • Stem cells in the pulp of primary teeth,
characterized as multipotent cells
• Helps in treatment of periodontal
disease, diabetes, spinal cord injury,
stroke, heart attack, burn, rheumatoid
arthritis and Parkinson's and
Alzheimer's, and regenerate many types
of tissue in the body.
Stem cells

27. TO REDUCE OROFACIAL PAIN
– Firstly, the modified adenovirus, adeno associated virus (AAV) or lipid
encapsulated plasmids coding for Interleukin-10, a therapeutic protein,
may be injected into the sub arachnoid space to transduce the pia mater
cells
– Secondly, a modified herpes virus may be introduced into the nerves of
Dorsal Root Ganglia (DRG) via an intradermal injection to the skin.
– In the DRG, it codes for an inhibitory neurotransmitter, an anti-
inflammatory peptide or decreases the synthesis of an endogenous
nociceptive molecule that results in alleviation of pain

28. IN CARCINOGENESIS..
• In 2003, the first gene therapy drug Gendicine, a recombinant
human adenovirus p53, was formulated and approved by the
Chinese authorities for the treatment of SCCHN and other
cancerous lesions.
• In 2005, the first genetically engineered oncolytic virus, H101
Adenovirus, was approved for treatment of SCCHN in China

29. MISSING TOOTH..
• A tooth germ can be created in vitro or ex vivo with a culture of
epithelial and mesenchymal stem cells. The tooth germ is
implanted into the alveolar bone to develop into a fully
functional, non-metal tooth implant.

30. Orthodontics in the year 2047 Genetically driven treatment plans
Havens, B. Wadhwa, S. and Nanda, R.:
J. Clin. Orthod. 41:549-556, 2007
Gene therapy for sutural growth disturbances:
• Mutations in FGFR2 have been linked to several human
craniosynostosis disorders, including Apert, & Crouzon syndromes.
• In cases of craniosynostosis involving mutations in FGFR2,
temporarily blocking FGFR2 signaling in the preosteoblasts within
the sutural mesenchyme or providing a different anti proliferation
signal to these cells would allow normal sutural growth without
surgical intervention.

31. Gene therapy to enhance condylar growth using rAAV-
VEGF(recombinant adeno associated virus- vascular
endothelial growth factor)
Objective: To test the hypothesis that the introduction of specific
vascular growth inducing genes would favorably affect
mandibular condyle growth in rats over a limited experimental
period
Result: Enhancement of mandibular condyle growth occurred in
backward and upward direction in VEGF group rather than
control group.

32. Local RANKL gene transfer to the periodontal tissue
accelerates orthodontic tooth movement
Kanzaki.H et al Gene Therapy 2006
Result –
It was demonstrated that transfer of the RANKL gene
to the periodontal-tissue ,activate osteoclastogenesis and
accelerated the amount of experimental Toot movement.
Local RANKL gene transfer might be a useful tool not
only for shortening orthodontic treatment, but also for moving
ankylosed teeth where teeth, fuse to the surrounding bone .

34. CONCLUSION
• Genetic Engineering is a vast field and accounts for
tremendous depth of knowledge yet to be understood.
• Many application of the Genetic Engineering applied in
various fields such as in Agriculture, Horticulture, Various
medications, vaccines productions etc has proved to be
beneficial
• There’s no doubt that this field will only improve the quality of
Life of Humans, But there still remains the question reliability
in the future.