Rapid and sensitive detection of various analytes is in high demand.Apart from its application in genome editing,CRISPR-Cas also shows promises in nucleic acid detection applications.To further exploit the potential o...Rapid and sensitive detection of various analytes is in high demand.Apart from its application in genome editing,CRISPR-Cas also shows promises in nucleic acid detection applications.To further exploit the potential of CRISPR-Cas for detection of diverse analytes,we present a versatile biosensing platform that couples the excellent affinity of aptamers for broad-range analytes with the collateral single-strand DNA cleavage activity of CRISPR-Cas12 a.We demonstrated that the biosensors developed by this platform can be used to detect protein and small molecule in human serum with a complicated background,i.e.,the tumor marker alpha fetoprotein and cocaine with the detection limits of 0.07 fmol/L and 0.34 lmol/L,respectively,highlighting the advantages of simplicity,sensitivity,short detection time,and low cost compared with the state-of-the-art biosensing approaches.Altogether,this biosensing platform with plug-and-play design show great potential in the detection of diverse analytes.展开更多
Clinical use of antimicrobials faces great challenges from the emergence of multidrug-resistant pathogens. The overexpression of drug efflux pumps is one of the major contributors to multidrug resistance(MDR). Reversi...Clinical use of antimicrobials faces great challenges from the emergence of multidrug-resistant pathogens. The overexpression of drug efflux pumps is one of the major contributors to multidrug resistance(MDR). Reversing the function of drug efflux pumps is a promising approach to overcome MDR. In the life-threatening fungal pathogen Candida albicans, the major facilitator superfamily(MFS) transporter Mdr1p can excrete many structurally unrelated antifungals, leading to MDR. Here we report a counterintuitive case of reversing MDR in C. albicans by using a natural product berberine to hijack the overexpressed Mdr1p for its own importation. Moreover, we illustrate that the imported berberine accumulates in mitochondria and compromises the mitochondrial function by impairing mitochondrial membrane potential and mitochondrial Complex I. This results in the selective elimination of Mdr1 p overexpressed C. albicans cells. Furthermore, we show that berberine treatment can prolong the mean survival time of mice with blood-borne dissemination of Mdr1p overexpressed multidrug-resistant candidiasis. This study provides a potential direction of novel anti-MDR drug discovery by screening for multidrug efflux pump converters.展开更多
基金supported by the National Natural Science Foundation of China (31770055, 31922002, 31720103901, and 31772242)the 111 Project (B18022)+4 种基金the Fundamental Research Funds for the Central Universities (22221818014)the Shanghai Science and Technology Commission (18JC1411900)the Young Scientists Innovation Promotion Association of Chinese Academy of Sciences (2016087) to Weishan Wangthe Shandong Taishan Scholar Program of China to Lixin Zhangthe Open Project Funding of the State Key Laboratory of Bioreactor Engineering
文摘Rapid and sensitive detection of various analytes is in high demand.Apart from its application in genome editing,CRISPR-Cas also shows promises in nucleic acid detection applications.To further exploit the potential of CRISPR-Cas for detection of diverse analytes,we present a versatile biosensing platform that couples the excellent affinity of aptamers for broad-range analytes with the collateral single-strand DNA cleavage activity of CRISPR-Cas12 a.We demonstrated that the biosensors developed by this platform can be used to detect protein and small molecule in human serum with a complicated background,i.e.,the tumor marker alpha fetoprotein and cocaine with the detection limits of 0.07 fmol/L and 0.34 lmol/L,respectively,highlighting the advantages of simplicity,sensitivity,short detection time,and low cost compared with the state-of-the-art biosensing approaches.Altogether,this biosensing platform with plug-and-play design show great potential in the detection of diverse analytes.
基金supported by the National Key Research and Development Program of China(2020YFA0907800,2022YFC3400200,and 2022YFA0912200)the National Natural Science Foundation of China(31900060)+1 种基金the Shanghai Pilot Program for Basic Research-Shanghai Jiao Tong University(21TQ1400204)the Natural Science Foundation of Shanghai(20ZR1414500)。
基金supported by the National Key Research and Development Program of China (2020YFA0907800)the National Natural Science Foundation of China (31720103901)+2 种基金the “111” Project of China (B18022)the Fundamental Research Funds for the Central Universities (22221818014S)the Open Project Funding of the State Key Laboratory of Bioreactor Engineering,the Shandong Taishan Scholar Award,and the Novo Nordisk Foundation (NNF10CC1016517)。
文摘Clinical use of antimicrobials faces great challenges from the emergence of multidrug-resistant pathogens. The overexpression of drug efflux pumps is one of the major contributors to multidrug resistance(MDR). Reversing the function of drug efflux pumps is a promising approach to overcome MDR. In the life-threatening fungal pathogen Candida albicans, the major facilitator superfamily(MFS) transporter Mdr1p can excrete many structurally unrelated antifungals, leading to MDR. Here we report a counterintuitive case of reversing MDR in C. albicans by using a natural product berberine to hijack the overexpressed Mdr1p for its own importation. Moreover, we illustrate that the imported berberine accumulates in mitochondria and compromises the mitochondrial function by impairing mitochondrial membrane potential and mitochondrial Complex I. This results in the selective elimination of Mdr1 p overexpressed C. albicans cells. Furthermore, we show that berberine treatment can prolong the mean survival time of mice with blood-borne dissemination of Mdr1p overexpressed multidrug-resistant candidiasis. This study provides a potential direction of novel anti-MDR drug discovery by screening for multidrug efflux pump converters.
