Genetic Transfection is a very useful and basic molecular biology technique of introducing nucleic acids into cells. In general terms, to transfect means to introduce genetic material (DNA, RNA, siRNA) into eukaryotic cells using chemical methods and without the use of viruses or electroporation machines.

1. Gene Transfection
(Cloning)

2. Introduction to Cloning
 Produce genetically identical copies of a
biological entity, DNA fragment
 Clones, organisms that are exact genetic
copies.
 Human Identical Twins.
 First cloned mammal was Dolly in 1996.

3. History
 1952: first experiment with frogs tails
 1970: Cloned frog develop into tadpole
 1984: Lamb born
 1994: Calves born,cloned from emryo of 120 cells
 1997:birth of Dolly
 1999: three goats cloned
 2000: Pigs cloned for use in transplantation
 2001: human embryo cloned not bigger 6 cells

4. Types of Cloning
Artificial
Embryo
Twinning
Natural Cloning
Gene cloning
Therapeutic
cloning
Reproductive
cloning
1. 2. 3.

5. Natural Cloning
 twins form, when the embryo splits in two.
 after egg and sperm join,
 resulting individuals are genetically identical.
 Natural clones, also known as identical twins
 Through asexual reproduction
 Both Identical twins have nearly the same genetic
makeup as each other
 but they are genetically different from either parent.

7. Gene Cloning
 A clone is exact copy of an organism, organ and
macromolecules.
 derived from the genome of an organism cloned into
vectors. An individual cell carrying a cloning vector.

10. Therapeutic Cloning
 creates embryonic Stem Cells.
 does not involve the creation of a perfectly copied
human being
 no sperm fertilization implantation into the uterus
 to grow healthy tissue to replace injured or diseased
tissues in the human body.
 This type of cloning usually use stem cells to treat
many diseases and many other problems related to
human health that encounter them.
 It does not make full human but make the human
organ.

12. Reproductive Cloning
 Production of genetic duplicate of existing
organism
 Somatic cell nuclear transfer
 Exact genetic copy
 This method used to create Dolly sheep

16. Gene transfer

17. Introduction
 Introduce foreign genes into the genome of target
cells.
 Directed desirable gene transfer from donor to
recipient
 The transferred gene is known as transgene
 The organism that develop after a successful
gene transfer is known as transgenic.

18. Methods of Gene transfer
 DNA transfer by natural methods
Transformation
Conjugation Transduction
Agrobacterium
mediated gene
transfer

19. DNA transfer by artificial methods
Microinjection
DNA transfer by
Calcium phosphate
methods

20. Transformation
 Direct uptake of exogenous DNA from its
surroundings
 occurs naturally in some species of bacteria,
 Cells that have undergone this treatment are said
to be competent.
 Any DNA that is not integrated into the
chromosome will be degraded.

22. Transduction
 Gene transfer from a donor to a recipient by way of a
bacteriophage.
 If the lysogenic cycle is adopted, the phage
chromosome is integrated into the bacterial
chromosome, where it can remain dormant for
thousands of generation
 The lytic cycle leads to the production of new phage
particles which are released by lysis of the host.

24. Conjugation
 process in which a recipient bacterium receives DNA
from a donor bacterium by cell-to-cell contact through
conjugative pili.
 Bacteria that have a F plasmid are referred to as F+
or male.
 Those that do not have an F plasmid are F- of
female.
 A conjugation event occurs when the male cell
extends his sex pili and one attaches to the female.

26. Agrobacterium mediated gene transfer
 A soil borne gram negative bacterium.
 It invades many dicot plants which are injured at
the soil level and causes crown gall disease.
 The ability to cause disease is associated with
the presence of the Ti plasmid within the
bacterial cell.
 Ti plasmid can be used to transport new genes
into plant cells.

28. Microinjection
 The microinjection is the process of transferring the
desirable DNA into the living cell ,through the use of
glass micropipette.

29. DNA transfer by Calcium phosphate methods
 This process involves the admixture of isolated DNA
with solution of calcium chloride and potassium
phosphate so precipitate of calcium phosphate to be
formed.
 Cells are then incubated with precipitated DNA either
in solution or in tissue culture dish.