To my knowledge there is no targeted gene insertion except 
for disruptions and maybe mutagenesis of a gene.
There is still the problem that e.g. insertion points of 
transgenes are rather difficult to determine (at least in 
our hands).
This is documented by different phenotypes that you may 
get with different independent clones. There is a lot of 
epigenetics going on and we have only scratched the 
surface.
Even with extrachromosomal vectors, phenotypes may vary. 
No idea why!
Correct me if I am wrong!
wolfgang


On Tue, 8 Oct 2013 22:44:35 -0400
  "John B. Biggins" <[log in to unmask]> wrote:
> Thanks.  I'm a chemist & I'm writing up some stuff & 
>need background. 
> 
> John
> 
> 
> On Oct 8, 2013, at 9:47 PM, Christopher West wrote:
> 
>> Yes, it is done all the time. You might start with:
>> 
>> Nat Protoc. 2007;2(6):1317-24.
>> Transformation of Dictyostelium discoideum with plasmid 
>>DNA.
>> Gaudet P, Pilcher KE, Fey P, Chisholm RL.
>> Source
>> dictyBase, Center for Genetic Medicine, Northwestern 
>>University, 676 North Saint Clair Street Suite 1260, 
>>Chicago, Illinois 60611, USA.
>> Abstract
>> DNA-mediated transformation is one of the most widely 
>>used techniques to study gene function. The eukaryote 
>>Dictyostelium discoideum is amenable to numerous genetic 
>>manipulations that require insertion of foreign DNA into 
>>cells. Here we describe two commonly used methods to 
>>transform Dictyostelium cells: calcium phosphate 
>>precipitation, resulting in high copy number 
>>transformants; and electroporation, an effective 
>>technique for producing single integration events into 
>>genomic DNA. Single integrations are required for gene 
>>disruption by homologous recombination. We also discuss 
>>how different selection markers affect vector copy number 
>>in transformants and explain why blasticidin has become 
>>the preferred selectable marker for making gene 
>>knockouts. Both procedures can be accomplished in less 
>>than 2 h of hands-on time; however, the calcium phosphate 
>>precipitation method contains several incubations, 
>>including one of at least 4 h, so the total time required 
>>for the transformation is approximately 8 h.
>> PMID: 17545968 [PubMed - indexed for MEDLINE]
>> 
>> -Chris West
>> 
>> 
>> On Oct 8, 2013, at 7:31 PM, "John B. Biggins" 
>><[log in to unmask]> wrote:
>> 
>>> Has anyone had any success in targeted insertion of 
>>>genes within the Dicty genome?
>>> 
>>> That is, you can stably insert the DNA you want in the 
>>>region you choose?
>>> 
>>> Thanks
>>> 
>>> John
>> 
> 

Wolfgang Nellen
Abt. Genetik
Univ. Kassel
Heinrich-Plett-Str. 40
34132 Kassel
Germany