Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brasil.
Departamento de Engenharia de Materiais, Escola de Engenharia de Lorena, Universidade de São Paulo, Lorena, SP, Brasil.
Braz J Med Biol Res. 2021 Apr 2;54(5):e10743. doi: 10.1590/1414-431X202010743. eCollection 2021.
Amphiphilic copolymers have a wide variety of medical and biotechnological applications, including DNA transfection in eukaryotic cells. Still, no polymer-primed transfection of prokaryotic cells has been described. The reversible addition-fragmentation chain transfer (RAFT) polymer synthesis technique and the reversible deactivation radical polymerization variants allow the design of polymers with well-controlled molar mass, morphology, and hydrophilicity/hydrophobicity ratios. RAFT was used to synthesize two amphiphilic copolymers containing different ratios of the amphiphilic poly[2-(dimethyl-amino) ethyl methacrylate] and the hydrophobic poly [methyl methacrylate]. These copolymers bound to pUC-19 DNA and successfully transfected non-competent Escherichia coli DH5α, with transformation efficiency in the range of 103 colony-forming units per µg of plasmid DNA. These results demonstrate prokaryote transformation using polymers with controlled amphiphilic/hydrophobic ratios.
两亲性共聚物在医学和生物技术领域有广泛的应用,包括真核细胞中的 DNA 转染。然而,目前还没有描述过用于原核细胞的聚合物引发的转染。可逆加成-断裂链转移(RAFT)聚合合成技术和可逆失活自由基聚合变体允许设计具有良好控制的摩尔质量、形态和亲水性/疏水性比的聚合物。使用 RAFT 合成了两种两亲性共聚物,它们含有不同比例的两亲性聚[2-(二甲氨基)乙基甲基丙烯酸酯]和疏水性聚[甲基丙烯酸甲酯]。这些共聚物与 pUC-19 DNA 结合,并成功地转染了非感受态的大肠杆菌 DH5α,其转化效率在每微克质粒 DNA 103 个集落形成单位的范围内。这些结果表明,使用具有控制的两亲性/疏水性比的聚合物可以进行原核转化。