Professor Takeo Miyake of Waseda University and senior researcher Tsutomu Mikawa of RIKEN have developed a nanosyringe that can introduce multiple proteins into cells with high efficiency and high survival rate.

 This research group has so far developed a composite nanotube sheet made of conductive polymer and metal, and developed a "nanosyringe" that can deliver substances to cells by inserting these nanotubes into cells.

 In this research, by improving this nanosyringe, we were able to introduce proteins, which had previously been difficult to deliver into cells, into cells with high efficiency and high survival rate.

 Using the developed technology, experiments were conducted to introduce lactate oxidase enzyme (LOx) into normal cells and cancer cells. The introduction efficiency of both methods was over 95%. While normal cells have a survival rate of over 100%, the lactic acid concentration inside cancer cells is about 10 times higher than in normal cells, so when LOx is introduced, hydrogen peroxide (a strong oxidant) is generated through an enzymatic reaction. It was confirmed that cancer cells were killed. Under conditions in which LOx was not delivered into cells, 24% of cancer cells remained after 33 hours, but when LOx was delivered into cells, up to 3% died.

 They also introduced a stable isotope-labeled protein (ubiquitin) into cells using a nanosyringe, and attempted NMR analysis using nuclear magnetic resonance spectroscopy (NMR). NMR analysis requires approximately 107 cells (1,000 million cells) into which a high concentration of protein has been introduced, but ubiquitin can be introduced into 1.8 x 107 cells using the nanosyringe system for NMR analysis. The company says it has demonstrated that it can be applied to

 In this way, with this technology, it is possible to introduce any functional protein into more than 1,000 million cells, which is the number of cells needed to handle in the field of regenerative medicine, with an introduction efficiency of 89.9% and a cell survival rate of 97.1%. We have confirmed that.

 In the future, we will use this technology to develop cell function modification (direct reprogramming) and intracellular function analysis (In-cell NMR), which is a method for analyzing biomolecules in living cells. The company also expects to develop the technology in cells other than cells (plants, yeast, lactic acid bacteria, etc.).

Paper information:【Analytical Chemistry】A Hybrid Nanotube Stamp system in Intracellular Protein Delivery for Cancer Treatment and NMR Analytical Techniques

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