Xueping Yang,Jingyun Li,Suixin Zhang,Chao Li,Jiehua Ma
CRISPR/Cas-based systems are highly attractive for developing next-generation diagnostic technologies because of their intrinsic merits such as simplicity, sensitivity, and specificity. However, currently, nucleic acid amplification procedures are still needed to achieve attomolar sensitivity in most CRISPR/Cas-based assays, which causes high cost, operation difficulty, and low efficiency. Herein, we combine the CRISPR/Cas12a-based assay and a single-microbead detection platform for one-step and amplification-free detection of DNA at the single-molecule level. By modifying DNA reporters on a biomimetic membrane-coated microbead, the activated Cas12a by targets will cleave these reporters and lighten the bead within 10 min. The method allows the detection of the target down to three copies in a 5 μL sample. Furthermore, we successfully apply this method for the specific identification of viral infection, foodborne bacteria, and DNA mutation in real samples without extra nucleic acid amplification. We believe that this approach offers new insights for developing CRISPR/Cas-based DNA assays in biomedical applications.
Su Yanbin,Sun Chengyu,Sun Xuwei,Wu Ruixue,Zhang Xing,Tu Yunzhou
The aim of this study was to explore whether or not acetylresveratrol as a potential substitute for resveratrol dragged the toxic aldehyde to inhibit the mutation of mitochondrial DNA. The results revealed that the acetylresveratrol shifted ultraviolet peak of trans -crotonaldehyde from 316 to 311 nm. In mitochondria, the acetylresveratrol split the ultraviolet peak at 311 nm of trans -crotonaldehyde into 311 nm and 309 nm; the aldehyde Raman band of trans -crotonaldehyde was red shifted by the acetylresveratrol from 1689 to 1686 cm −1 with obvious band decline; Raman bands at 1149 cm −1 , 1168 cm −1 , and 1325 cm −1 of acetylresveratrol disappeared. In aldehyde dehydrogenase, the aldehyde Raman band of trans- crotonaldehyde was red shifted by the acetylresveratrol from 1689 to 1684 cm −1 with band decline; Raman bands at 1150 cm −1 , 1168 cm −1 , and 1324 cm −1 of acetylresveratrol declined. The weak acidic microenvironment was the best, for the acetylresveratrol dragged the toxic aldehyde of trans -crotonaldehyde. Compared with the resveratrol, the effect of the acetylresveratrol on the toxic aldehyde of trans -crotonaldehyde was very similar to that of the resveratrol. The acetylresveratrol is very suitable as a potential substitute for resveratrol dragged the toxic aldehyde to inhibit the mutation of mitochondrial DNA.
In mitochondria, the Raman band of the toxic –CH=O of trans- crotonaldehyde (TCA) dragged by the Acetyl-Res from 1689 to 1686 cm −1 with obvious band decline, while the Raman bands at 1149 cm −1 , 1168 cm −1 , and 1325 cm −1 of the Acetyl-Res disappeared, respectively. The Acetyl-Res is very suitable as a potential substitute, for the Res dragged the toxic –CH=O of TCA to inhibit the mutation of mitochondrial DNA for anticancer.
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