2results about How to "Enhance electrostatic interaction" patented technology

This invention discloses an engineered mutant CpAgo_PM based on a novel PAZ nucleic acid binding pocket and its applications, belonging to the fields of molecular biology and genetic engineering. The amino acid sequence of the mutant CpAgo_PM is shown in SEQ ID NO.1. This invention rationally designs the PAZ nucleic acid pocket, mutating some normally non-positively charged amino acid residues in the pocket to positively charged amino acids (CpAgo_PM). In in vitro cleavage experiments, four different target sites (TET2-1 / 2 / 3 / 4) on the human TET2 gene transcript were tested. Compared with CpAgo_WT, CpAgo_PM significantly improves the cleavage efficiency at three sites: TET2-1, TET2-2, and TET2-4, indicating that it partially overcomes the sequence cleavage bias of the wild type. Furthermore, its gene editing function in a eukaryotic cell environment has been clearly demonstrated.

The application belongs to the field of engineering technology and provides a preparation method of a nucleic acid separation device. The preparation method of the application prepares a hydrogel nucleic acid adsorption device through a photo-initiated radical polymerization reaction, avoiding the cumbersome limitations of traditional polymerization reactions which need to be carried out under anaerobic conditions. The preparation method is simple and fast, and does not depend on professional and precise instruments. The raw materials are cheap and easy to obtain. The method can efficiently load silica particles, effectively combine the plasticity of the hydrogel with the high specific surface area of the silica particles and the strong binding capacity of the nucleic acid, and further obtain a handheld nucleic acid extraction device which can change shape, size and quantity at any time according to application requirements, has a high specific surface area and excellent nucleic acid extraction performance. Compared with traditional processes, the preparation process is simpler, the application range is wider, and the application requirements of low resource environment are greatly met, and the application prospect in the field of outdoor and clinical nucleic acid extraction technology is broad.