Biopolym. Cell. 2022; 38(1):58-68.
http://dx.doi.org/10.7124/bc.000A73
Bioorganic Chemistry
Transformation of the moss (Ceratodon purpureus) with plasmid DNA delivered by novel block-copolymers of the dimethylaminoethyl methacrylate
1, 2Finiuk N. S., 3Mitina N. E., 4Lobachevska O. V., 3Zaichenko A. S., 1, 2Stoika R. S.
  1. Institute of Cell Biology, NAS of Ukraine
    14/16, Drahomanov Str., Lviv, Ukraine, 79005
  2. Ivan Franko National University of Lviv
    4, Hrushevskoho Str., Lviv, Ukraine, 79005
  3. Lviv Polytechnic National University
    12, S. Bandery Str., Lviv, Ukraine, 79013
  4. Institute of Ecology of the Carpathians of National Academy of Sciences of Ukraine
    4, Kozelnytska Str., Lviv, Ukraine, 79026

Abstract

Aim. To investigate the potential of poly(2-dimethylamino)ethyl methacrylate (DMAEMA)-based block-like polymers to serve as gene delivery carriers in moss Ceratodon purpureus (Hedw.) Brid. protoplasts, and to evaluate the level of their phytotoxicity. Methods. Organic synthesis; DNA gel retardation assay; adapted PEG-mediated transformation protocol; PCR; light microscopy. Results. The formation of pDNA complex with DMAEMA-based carriers took place at 0.01-0.1 % concentrations of the polymer. The poly-DMAEMA carriers F8-DM1, F8-DM2 (fluorine-containing), LAcr-DM1, LAcr-DM2 (lauryl acrylate-containing), BAcr-DM1, and BAcr-DM2 (butyl acrylate-containing) were effective as carriers of plasmid DNA pSF3 at C. purpureus transformation. PCR analysis confirmed that the transformants of C. purpureus moss contain GFP as a gene of interest after the protoplast transformation by polymers LAcr-DM2, LAcr-DM1, BAcr-DM2, BAcr-DM1 and F8-DM2. The poly-DMAEMA carriers at working concentration (0.0025 %) were relatively non-toxic for protoplasts of C. purpureus moss. 83.1-93.9 % of viable protoplasts of C. purpureus moss were found after the treatment with studied carriers at that dose. However, at 0.25 % i.e. 100 times higher concentration than that used for moss transformation, the poly-DMAEMA carriers reached their IC50 level. Conclusion. The novel block-like poly-DMAEMA carriers were effective in transformation of C. purpureus moss protoplasts and demonstrated low toxicity.
Keywords: polymeric carrier, poly(2-dimethylamino)ethyl methacrylate, moss Ceratodon purpureus, protoplasts transformation, toxicity.
Full text: (PDF, in English)

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