DNA integration after transfection of mammalian cells

kang at msvax.mssm.edu kang at msvax.mssm.edu
Fri Mar 3 01:32:53 EST 1995


In article <3j5qhi$7hp at quandong.itd.adelaide.edu.au>, Ian Lyons <ilyons at biochem.adelaide.edu.au> writes:
>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
>
>IAN LYONS
>Biochem, Adelaide
>AUSTRALIA
>

Late DNA integration theory of pronucleus injected DNA id interesting.
Can you give me the reference? appreciated.

If you can not find Eppendorf injector you can have Nicon or Picospiritzer II
It works fine.

Chulho Kang




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