Hi!
Does anybody have any idea of how mutagenic is biolistic transformation?
thanks!
[b]“Targeted homologous recombination is a powerful approach for genome manipulation that is widely used for gene alteration and knockouts in mouse and yeast.
In Caenorhabditis elegans, several methods of target‐selected mutagenesis have been implemented but none of them provides the opportunity
of introducing exact predefined changes into the genome.
Although anecdotal cases of homologous gene targeting in C.elegans have been reported, no practical technique of gene targeting has been developed so far.
In this work we demonstrate that transformation of C.elegans by microparticle bombardment (biolistic transformation) can result in homologous recombination
between introduced DNA and the chromosomal locus.
We describe a scaled up version of biolistic transformation that can be used as a method for homologous gene targeting in the worm.”
Received January 8, 2004; Revised and Accepted January 22, 2004
Oxford University Press[/b]I think that it’s hard to get the mutation rate (though maybe people who’ve done more bombardment might be able to give you a ballpark estimate). I figure it’s hard to tell since you’re bombarding unc-119s and looking for animals rescued for the unc and dauer exit phenotypes. You can’t look for dpys and uncs like one might for an EMS screen and infer mutation rate that way.
My guess would be mildly mutagenic? Since the process relies on the occasional gold particle hitting a germline or oocyte nucleus and creating a DSB, many should be repaired by HR, but the occasional DSB would probably be repaired with errors.