MIDGE (minimal immunologically defined gene expression) DNA最小免疫限定基因表達(dá)載體

Viral and plasmid vectors may cause immunological side-effects resulting from the expression of therapeutically unwanted genes and from CpG motifs contained in their sequence. A new vector type for minimalistic, immunological-defined gene expression (MIDGE) may overcome these problems. BioVector's MIDGE is a minimal size gene transfer unit consisting of the expression cassette, including promotor, gene and RNA-stabilizing sequences, flanked by two short hairpin oligonucleotide sequences. DNA not encoding the desired gene is reduced to a minimum. To compare transfection efficiencies in vivo hydrodynamics-based, systemic transfection was performed in BALB/c mice with BioVector's MIDGE vectors and corresponding plasmids. The transfection efficiencies of the BioVector's MIDGE vectors as measured by luciferase expression were significantly higher in liver (2.5-fold), lung (3.5-fold), kidneys (3.9-fold) and heart (17-fold) as compared to plasmids. The mean numbers of BioVector's MIDGE vector molecules per cell as measured by quantitative PCR were also significantly higher. These advantages suggest the preferential use of this new vector type for clinical gene therapy studies.

BioVector's MIDGE DNA

Minimized DNA
in linear form with caps
BioVector's MIDGE vector features
? no bacterial backbone sequences
(e.g. ori, antibiotic resistance marker)
? linear DNA fragment of minimal size
? highly purifi ed
? reduced CpG content
? improved safety
? minimized side effects

An alternative concept for the production of minimal DNA vectors, which contain only the information necessary for the intended effect, is e.g. BioVector's MIDGE (minimal immunologically defined gene expression) DNA. The BioVector's MIDGE structure is linear and tightly closed by single-stranded hairpin regions at both ends. This leads to the characteristic small size of these molecules, which – like the minicircles – have already been used in preclinical research as well as in clinical studies.

However, such linear molecules do not have the superhelicity characteristic of intact plasmids and PlasmidFactory's minicircles, so that minicircle DNA is, in our view, the preferred system when choosing a suitable DNA vector.

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