期刊
ACS APPLIED MATERIALS & INTERFACES
卷 12, 期 52, 页码 57695-57709出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsami.0c16491
关键词
Multiplex detection; versatile; normalized signal; signal amplification; mesoporous silica nanoquenchers
资金
- GSK-EDB Trust Fund [R-143-000688-592]
- MOE-T1 of Singapore [R-143-000-694-114]
- National Natural Science Foundation [31900982, 52,072,418]
- Science and Technology Innovation Foundation of Shenzhen [JCYJ20190807151807459]
- Fundamental Research Funds for the Central Universities of China [19ykpy142]
Detection of endogenous tumor-related RNA is vital for cancer diagnostics. Despite advancements made, live-cell RNA detection still faces numerous problems, such as low signal output and cell-to-cell variations arising from differences in probe uptake. To address these issues, we designed a versatile and highly sensitive mRNA/miRNA nanosensor featuring, for the first time, signal amplification and in-built signal normalization. Using dye-loaded mesoporous silica nanoquenchers (qMSNs) capped with target-corresponding antisense oligos (ASOs), direct fluorescence Turn-ON with signal amplification was achieved upon target binding. By readily varying the capping ASOs as well as cargo dyes, a suite of RNA nanosensors for multiplex target detection could be easily prepared. Further modification of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA-responsive molecular beacons (MBs) onto our nanosensor enabled dual detection of target RNA and GAPDH mRNA, allowing for target signal normalization using GAPDH as a reference. We demonstrated that this newly developed nanosensor could successfully differentiate between noncancer and cancer cells, as well as accurately monitor the relative expression levels of multiple tumor-related RNAs simultaneously in different cancer cell lines, with a high degree of specificity and sensitivity, functioning as a noninvasive qPCR mimic imaging tool in live cells.
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