Functionalisation of calcium phosphate nanoparticles by oligonucleotides and their application for gene silencing
Functionalisation of calcium phosphate nanoparticles by oligonucleotides and their application for gene silencing
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DOI：
10.1039/b612699e
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发表时间：
2007-01-01
影响因子：
--
通讯作者：
Epple, Matthias
中科院分区：
文献类型：
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作者：
Sokolova, Viktoriya;Kovtun, Anna;Epple, Matthias
在分子生物学中，蛋白质的产生可以通过将特定的寡核苷酸引入活细胞来有效地抑制（基因沉默或反义策略;对基因治疗很重要）。磷酸钙纳米颗粒由于其高度的生物相容性和生物降解性，可以在这种治疗应用中用作生物分子的载体。通过用单链和双链寡核苷酸涂覆无机纳米颗粒来制备稳定的胶体。通过动态光散射、zeta电位测量、透射电子显微镜和扫描电子显微镜对分散体进行分析。在优化的条件下得到了粒径约为100 nm的纳米粒子。在其绿色荧光被涂覆的纳米颗粒（用siRNA进行基因沉默）关闭的HeLa-EGFP细胞上测试这种纳米颗粒特异性抑制蛋白质合成的效率。如果将siRNA掺入磷酸钙颗粒中，从而防止细胞内降解，则转染效率显著增加。该分散体是稳定的，可以在4 ℃下储存数周而不丧失活性，使其可用作生化试剂。
In molecular biology, the production of proteins can be effectively inhibited by introducing specific oligonucleotides into a living cell ( gene silencing or antisense strategy; important for gene therapy). Calcium phosphate nanoparticles can serve as carriers for biomolecules in such therapeutic applications due to their high biocompatibility and biodegradability. Stable colloids were prepared by coating the inorganic nanoparticles with single- and double-stranded oligonucleotides. The dispersions were analysed by dynamic light scattering, zeta potential measurements, transmission electron microscopy, and scanning electron microscopy. Particles with a diameter of about 100 nm were obtained under optimized conditions. The efficiency of such nanoparticles to specifically inhibit protein synthesis was tested on HeLa-EGFP cells whose green fluorescence was turned off by the coated nanoparticles ( gene silencing with siRNA). If siRNA was incorporated into the calcium phosphate particle and thereby protected from intracellular degradation, the transfection efficiency was significantly increased. The dispersions were stable and could be stored at 4 degrees C without loss of activity for several weeks, making them available as biochemical reagents.