Scientists invent a new method of RNA interference experiment is simple and accurate for high-throughput analysis
In the latest "Nature-Methodology" (Nature Methods) magazine, Xie Xiaoliang's research group published an article again, proposing the use of its research and development technology-stimulated Raman scattering (stimulated Raman scattering (SRS) microscopy technology A new method for conducting RNA interference experiments.
This new method uses stimulated Raman scattering microscopy without fluorescence labeling for quantitative imaging, which overcomes the problems of chromatographic analysis such as cumbersome biochemical analysis methods, and the high background signal of CARS microscopy methods and inaccurate results. A simple and accurate new technology suitable for high-throughput analysis. In this article, the researchers used this method to find eight new regulatory factors in the regulation of fat storage.
Obesity is a major illness in modern society, which can cause various chronic diseases, such as type 2 diabetes, cardiovascular disease and so on. In order to better understand obesity and its related metabolic disorder, we need to deeply analyze the regulatory mechanism of fat accumulation at the cellular and tissue levels. At present, genome-wide analysis of some model organisms ’fat storage regulators has been carried out, but these Not enough.
The general analysis methods of nematode fat storage regulators mainly include traditional biochemical analysis methods, such as fixed staining of nematodes, fluorescence imaging, etc. Recently, some research groups have reported the use of CARS microscopy technology for analysis. However, these two methods have some limitations. Biochemical analysis methods need to extract fat from nematodes and perform tedious chromatographic analysis. This is impossible at the genomic level, and some organic solvents used can also interfere with experimental analysis. . The analysis using CARS microscopy has the problems of high background signal and limited sensitivity.
The SRS microscopy technique applied in this article can quickly respond to the laser and precisely modulate to remove background noise, so that not only can the same spectrum as the traditional Raman spectroscopy be obtained, but also the signal intensity is several orders of magnitude higher. Using this method combined with RNAi screening, researchers can search for genes that regulate fat storage under physiological conditions. After experimental analysis, the researchers found 9 key regulatory factors from 272 genes.
Combining SRS microscopy technology with RNAi technology can help researchers observe the regulation of fat metabolism under the active physiological conditions of nematodes, and this method does not require labeling, which greatly simplifies the sample preparation process and is suitable for cell tracking of model organisms , Also conducive to high-throughput screening analysis.
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