@entity4210 is an obligate intracellular bacterium that causes the @entity281 , granulocytic anaplasmosis. While electroporation can transform A. phagocytophilum isolated from host cells, no method has been developed to transform it while growing inside the ApV (A. phagocytophilum-occupied vacuole). @entity21454 ( @entity21455 ) dendrimers, well-defined tree-branched macromolecules used for gene therapy and nucleic acid delivery into @entity1 cells, were recently shown to be effective in transforming @entity1047 spp. actively growing in host cells. We determined if we could adapt a similar system to transform A. phagocytophilum. Incubating fluorescently labeled @entity21455 dendrimers with infected host cells resulted in @entity941 -positive ApVs. Incubating infected host cells or host cell-free A. phagocytophilum organisms with dendrimers complexed with pCis GFPuv-SS Himar A7 plasmid, which carries a Himar1 transposon cassette encoding GFPuv and @entity6760 / @entity3639 resistance plus the Himar1 transposase itself, resulted in GFP-positive, antibiotic resistant bacteria. Yet, transformation efficiencies were low. The transformed bacterial populations could only be maintained for a few passages, likely due to random Himar1 cassette-mediated disruption of A. phagocytophilum genes required for fitness. Nonetheless, these results provide proof of principle that dendrimers can deliver exogenous DNA into A. phagocytophilum, both inside and outside of host cells.

Dendrimer-enabled transformation of XXXX .

@entity1 :: ('9606', 'Species') :: ['mammalian'] @entity21454 :: ('MESH:C531249', 'Chemical') :: ['Polyamidoamine'] @entity3639 :: ('MESH:D013307', 'Chemical') :: ['streptomycin'] @entity281 :: ('MESH:D007239', 'Disease') :: ['emerging infection'] @entity941 :: ('MESH:D019793', 'Chemical') :: ['fluorescein'] @entity21455 :: ('CHEBI:53199', 'Chemical') :: ['PAMAM'] @entity1047 :: ('MESH:D002690', 'Disease') :: ['Chlamydia'] @entity6760 :: ('MESH:D000198', 'Chemical') :: ['spectinomycin'] @entity4210 :: ('948', 'Species') :: ['Anaplasma phagocytophilum']
@entity4210 :: (948,Species) :: ['Anaplasma phagocytophilum']
