Transformation of the moss (Ceratodon purpureus) with plasmid DNA delivered by novel block-copolymers of the dimethylaminoethyl methacrylate

Q4 Biochemistry, Genetics and Molecular Biology

Biopolymers and Cell Pub Date : 2022-07-25 DOI:10.7124/bc.000a73

N. Finiuk, N. Mitina, O. Lobachevska, A. Zaichenko, R. Stoika

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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 IC 50 level. Conclusion. The novel block-like poly-DMAEMA carriers were effective in transformation of C. purpureus moss protoplasts and demonstrated low toxicity.

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新型甲基丙烯酸二甲氨基乙酯嵌段共聚物介导的质粒DNA转化苔藓（Ceratodon purpureus）

目标研究基于聚（2-二甲氨基）甲基丙烯酸乙酯（DMAEMA）的嵌段状聚合物作为紫苔基因递送载体的潜力。原生质体，并评估其植物毒性水平。方法。有机合成；DNA凝胶阻滞试验；适用的PEG介导的转化方案；PCR；光学显微镜。后果pDNA复合物与基于DMAEMA的载体在0.01-0.1%的聚合物浓度下形成。聚DMAEMA载体F8-DM1、F8-DM2（含氟）、LAcr-DM1、LAcr-D-M2（含丙烯酸月桂酯）、BAcr-DM1和BAcr-DM2（含有丙烯酸丁酯）在紫曲霉转化中作为质粒DNA pSF3的载体是有效的。PCR分析证实，在通过聚合物LAcr-DM2、LAcr-DM1、BAcr-DM2，BAcr-DM1和F8-DM2进行原生质体转化后，紫苔的转化体含有GFP作为感兴趣的基因。在工作浓度（0.0025%）下，聚DMAEMA载体对紫苔原生质体相对无毒。在用该剂量的所研究的载体处理后，发现83.1-39.9%的紫苔C.purpureus moss的活原生质体。然而，在0.25%，即比用于苔藓转化的浓度高100倍的浓度下，聚DMAEMA载体达到其IC50水平。结论新型嵌段型聚DMAEMA载体对紫苔原生质体的转化效果良好，且具有低毒性。

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来源期刊

Biopolymers and Cell Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)

CiteScore

1.10

自引率

0.00%

发文量

期刊介绍： “Biopolymer and cell” is published since 1985 at the Institute of Molecular Biology and Genetics NAS of Ukraine under the supervision of the National Academy of Sciences of Ukraine. Our journal covers a wide scope of problems related to molecular biology and genetics including structural and functional genomics, transcriptomics, proteomics, bioinformatics, biomedicine, molecular enzymology, molecular virology and immunology, theoretical bases of biotechnology, physics and physical chemistry of proteins and nucleic acids and bioorganic chemistry.