Methods of genetic modification principles and applications

1. M.SC/BCH/20/1165

2. Explanation
of Genetic
Modification
Methods of
Genetic
Modification
Description
of the
methods
Advantages
And
limitations
of each
Method

3.  Genetic modification is the process of altering
the genetic makeup of an organism. This has
been done indirectly for thousands of years
by controlled, or selective, breeding of plants
and animals.
 Modern biotechnology has made it easier and
faster to target a specific gene for more-
precise alteration of the organism through
genetic engineering.

4.  Genetic modification dates back to ancient times, when humans influenced
genetics by selectively breeding organisms, according to article by Gabriel
Rangel, public health scientist at Harvard University When repeated over
several generations, this process leads to dramatic changes in the species.
 The history of genetic alteration as a technical science begins in the 1970s.
The history of genetic manipulation originated with the first recombinant
DNA being produced. A plethora of engineering teams worked on this
project. However, only two researchers, Boyer and Cohen, were successful.
They combined their common knowledge of DNA and cellular
reproduction to produce the first recombinant DNA.

5.  Methods of genetic modification are divided
into two
1. Traditional Method of Genetic modification
And
2. New Method of Genetic modification or
Genetic Engineering

6.  There are three traditional methods of genetic
modification:
1. Simple selection
2. Crossing
3. Mutant isolation

7.  Traditional techniques for gene modification
limited modifications to those occurring
between closely related organisms.
 Non desired trait (genes) can be transferred
 Time consuming

8.  There are three methods of new genetic
modification or genetic Engineering
 Plasmid Method
 Vector Method
 Biolistic Method

9.  Plasmid method
is the most
common used
method of
altering the
genes of any
organism.
Bacterial genetic
variation takes
place through
this method.
Plasmid method
occur in the
following ways

10.  It eliminates the need to inject infectious
agents.
 Plasmid stimulates both B- and T-cell immune
responses.
 Plasmids are more stable in different
temperatures so are easier to store and
transport.
 Plasmids are also easy to construct and can be
inexpensively produced in large quantities
 Detection is easier because of antibiotic-
resistant genes.

11.  Traditionally, plasmid manufacture
involves E. coli fermentation, which is
slow, expensive, and limited by lack of
capacity.
 Batch failure are common in plasmid
method.
 Antibiotic resistance in the final products

13.  High gene transduction efficiency
 Transgene expression can be controlled
by virus(transient expression or
persistent expression
 The method can target specific cells
types such as dividing cells or non
dividing cells

14.  Viral vector are difficult to
manufacture or produce in low titers
 Immune reaction
 Limitation in packaging capacity e.g
4.5kb for AAV vector
 May lead to insertion of mutagenesis

15.  DNA-coated metal
particles are placed
on the front end of
a bullet. High-
pressured gas
drives the bullet
down a shaft. At
the end of the
shaft, the bullet is
blocked, but the
DNA-coated
particles emerge
with great speed
and force.


16.  Method is easy to use, rapid and versatile
 Transient or stable expression is possible
 No carrier DNA is needed and no extraneous
genes or proteins are delivered
 Co-delivery of multiple plasmids and high
levels of co-transformation are possible
 Large DNA fragments may be transferred as
well as small interfering RNA's for gene
silencing
 Many cells types can be transfected,
including non-dividing cells and plants
 Potentially toxic treatments such as using
viruses or chemical are avoided

17.  Causes cellular damage, for example when
pulse are of wrong length or intensity it may
cause pores of cell becomes too large or
causing cell damage or rupture
 Ion imbalance that could lead to improper
cell function and cell death.
 Frequency of transformation is low
 Integration is low
 Requirement of equipment and skill personal
is require

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