The standards for transformation and transfection for years have been plasmid transfer by conjugate mating, DNA uptake by competent cells, chemical permeation, electroporation, and detergent and liposomal vectors and viral vectors, of bacterial and eukaryotic cells, respectively. The new standards for genetic modification employing such vectors involve CRISPR which causes directed double-strand DNA breaks and homologous recombination/repair (the latter using endogenous cellular processes). Off target changes can occur, although recent efforts have been made by the developers to avoid such untoward effects.
BioSPLICE, BIOSYNTHETIC PLASMID INTEGRATED CAPTURE ELEMENTS, US Patent 8,628,955 B2, Jan. 14, 2014, was designed to provide a new synthetic to biosynthetic vector which avoided these issues and could be used in vivo as well as in vitro due to the lack of protein and subsequent host immune response (unlike viral vectors). Furthermore, the vector can be made in vitro by purely chemical means or in vivo using genetically modified cells. The process has a built-in fail safe system because DALM can absorb microwave to radiofrequency radiation, high enough levels leading to destruction of the vector or the cell which contains the vector or is biosynthesizing more DALM and subsequent vectors. Cross strain and cross species biosynthetic production of transforming or transfecting vectors in up to three iterations (biosynthesis by subsequently transformed bacterial cells and the further transformation of other bacteria with the new vectors has been demonstrated, for three “generations”). These bacterial vectors have also transfected human and animal cells. The horizontal transfer from eukaryotic cell to eukaryotic cell has not been demonstrated yet but the production of nanoparticle forms of DALM by these cell has. These transfers of genetic material by the DALM vector yields stable transformants and transfectants, as well as those capable of producing chemically functional DALM in prokaryotes and eukaryotes. The DALM vectors have also demonstrated transfer, by PCR, of small synthetic nucleic acids, DNA aptamers with primers, to eukaryotic cells.
The intent is to expand this approach to synthetic expressible micro-linear DNA. The DALM nanoparticles have been combined with magnetic ferric nanoparticles due to the natural chelating of metals by the DALM (especially useful in magnetically capturing and purifying the DALM vectors). The DALM nanoparticles have also been combined with other metals such as silver and nickel for destruction of microbial targets and also with carbon nanotubes and quantum dots to trace their locations in transfected target cells. Besides laboratory investigations, the DALM vectors have been applied to pathogens like anthrax bacteria and demonstrated direct-mediated microwave destruction and genetic transformation to change antibiotic-sensitivity and pathogenic properties in anticipation of addressing selective destruction of antibiotic resistant microbes and attacking pathogenic genetic islands in the genomes of pathogens. DALM vectors can accomplish these tasks because DALM readily interacts and binds tightly with DNA but retains the specific binding, expression, PCR, and homologous recombination capabilities of the bound DNA. The complete history of the DALM vector R&D can be found in The Black Dragon Trilogy published at Amazon.com as an eBook.