DNA integration after transfection of mammalian cells
ilyons at biochem.adelaide.edu.au
Thu Mar 2 20:14:26 EST 1995
DNA injected into fertilised mouse eggs does not seem to integrate
until the four to 16 cell stage. This means that at the blastocyst
stage, almost all injected pronuclear eggs have developed to
chimaeras. That only a small number of Inner Cell Mass cells
go on to form the embryo proper is probably what reduces the
proportion of chimaeric live young to the numbers we see.
It's sort of a cellular founder effect.
Is this the case for cultured cells? Do only a proportion
of the daughter cells of a transfected cell get transformed?
In most experiments in which we apply selection immediately
after transfection we only see the progeny of cells which
integrated the DNA.
Does integration occur immediately, or 0 to 3 cell divisions
after DNA introduction?
Facts or theory welcome!
Any suggestions as to the best microinjection device to do
this experiment with? Do Eppendorf still make a microinjector
driven by compressed gas?
Thanks for any input
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