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Gene
transfection
General Troubleshooting
An introduction to transfection methods
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Gene transfection means the delivery and introduction of biologically functional nucleic acid into a cell,
by
which the nucleic acid retains its function within the cell. The nucleic acid includes DNA (plasmid and
linear
double
strand DNA), antisense oligonucleotide and RNAi (RNA interference). Gene transfection
has been widely
applied in
genomic function studies (gene regulation, gene function, signal transduction
and drug screen studies)
and gene
therapy studies.
Gene carrier is needed for introducing nucleic acid into cells. Both viral and non-viral gene carrier have
been used
in gene transfer. Viral vectors have highest efficiency, but the safety of virus, the high expense
and the
complicated procedure in viral vector preparation hampers its applicability. On the other hand,
nonviral transfection
reagent is needed to deliver viral DNA into cell during viral vector preparation.
Gene transfection reagent is needed to deliver the target gene into a cell during gene transfection.
Calcium
phosphate shows low transfection efficiency, and is of no effect for a large number of cell lines,
so it doesn't meet
the most needs in this field. Presently the most popular gene transfection reagent is
cationic lipids and cationic
polymers. Both of them can overcome the cellular barriers and carry nucleic acid
into cell. Cationic lipids show high
transfection efficiency in vitro gene delivery. However, they are not
suitable for in vivo administration, because they
will be rapidly cleared by the plasma, moreover they can
accumulate within the lung tissue and induce potent anti-inflammatory activity in vivo, which will induce
high level of toxicity. Owing to above limitation of cationic lipid, there
is a growing interest in cationic polymer
gene carriers.
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