Hayashi Mirian A F, Oliveira Eduardo B, Kerkis Irina, Karpel Richard L
Departamento de Farmacologia, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP, Brazil.
Methods Mol Biol. 2012;906:337-52. doi: 10.1007/978-1-61779-953-2_28.
Crotamine is a basic, 42-residue polypeptide derived from snake venom that has been shown to possess cell-penetrating properties. Crotamine forms nanoparticles with a variety of DNA and RNA molecules, and crotamine-plasmid DNA nanoparticles are selectively delivered into actively proliferating cells in culture or in mice. As such, these nanoparticles could form the basis for a nucleic acid drug-delivery system. Here we describe the preparation, purification, and biochemical and biophysical analysis of venom-derived, recombinant, chemically synthesized, and fluorescent-labeled crotamine; the formation and characterization of crotamine-DNA and -RNA nanoparticles; and the delivery of these nanoparticles into cells and animals.
巴曲酶是一种源自蛇毒的碱性42肽,已被证明具有细胞穿透特性。巴曲酶能与多种DNA和RNA分子形成纳米颗粒,且巴曲酶-质粒DNA纳米颗粒能被选择性地递送至培养的或小鼠体内的活跃增殖细胞中。因此,这些纳米颗粒可为核酸药物递送系统奠定基础。在此,我们描述了源自毒液的、重组的、化学合成的及荧光标记的巴曲酶的制备、纯化、生化及生物物理分析;巴曲酶-DNA和-RNA纳米颗粒的形成及表征;以及这些纳米颗粒向细胞和动物体内的递送。