CaPO transfection theories?

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Mon Mar 22 12:45:00 EST 1999


:Hello, anyone have any theories on the mechanism of the Calcium Phosphate
:coprecipitation transfection method? Nobody seems to know how it works, only
:that it works !
:thanks,
:dk

There is no one single paper that offers unequivocal proof of it but
there is a lot of circumstantial evidence suggesting it is taken up
by endocytosis (dependence on ppt size, enhancement by 
inhibitors of lysosomal function, punctate pattern of staining, 
large extent of degradation). The tiny faction that escapes hydrolysis
due to protection in ppted form, enters nucleus, likely still in some
form of precipitate (hence significant extent of multicopy tandem 
inserts). 

On the other hand, it is interesting to note that mechanisms of 
Ca2+-mediated E.coli transformation, natural transformation of 
most becteria, and even exact mechanism of Lambda and T4 
DNA penetration through membrane are still largely unknown.
The sad thing about it is that nobody appears to care anymore. 

At least we know mechanism of electrotransfection and 
electroporation... :-))

        - Dima





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