Design and screening of RNA fluorescent aptamers based on broccoli
In order to provide more precise tools for RNA imaging in living cells,this paper aims to create green fluorescent aptamers with high fluorescence intensity and good stability based on broccoli's RNA fluo-rescent labeling.The broccoli was optimized by rational design,and its variants were screened for multiple rounds by measuring spectral characteristics and physicochemical properties.The results showed that based on three rounds of mutation screening of the RNA fluorescent aptamer broccoli,a broccoli four-site mutant(named Bro)with a fluorescence intensity increase of 3.3 times was obtained;The 3WJ-n×Bro RNA fluo-rescent aptamer family was established by using the designed 3WJ scaffold.The fluorescence intensity on the aptamer can be significantly improved by embedding tandem repeat Bro,with the fluorescence intensity of 3WJ-4×Bro is 2.7 times higher than that of 3WJ-Bro;It also shows lower salt ion dependence and higher thermal stability,light stability and monitoring sensitivity.In this paper,we designed the 3WJ-n×Bro family and screened out the excellent aptamer 3WJ-4×Bro,which provided a visualization tool for live cell RNA im-aging at the RNA level.
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