Published online Dec 26, 2021. doi: 10.4252/wjsc.v13.i12.1918
Peer-review started: June 18, 2021
First decision: July 17, 2021
Revised: August 11, 2021
Accepted: December 10, 2021
Article in press: December 10, 2021
Published online: December 26, 2021
Although mRNA analysis is still conventionally achieved by fluorescence in situ hybridization and qRT-PCR, there is a strong need for real-time monitoring of specific RNA transcripts in living cells, both for a qualitative and quantitative assessment. Within this context, SmartFlareTM technology is a reliable tool for evaluating the presence and the upregulation/downregulation of mRNAs in individual living cells. In addition, this nanotechnology offers the advantages of retaining cell viability, morpho-functional and biochemical properties and allowing downstream experiments.
SmartFlareTM technology is a devoted and straightforward method for the spati
To study the dynamics of differentiation-related RNA transcripts in human neural stem cells.
The presence of CD133 and OCT4 mRNA-linked nanoprobes in neurosphere-derived cells (from 3 to 30 DIV) was investigated by SmartFlareTM as a reliable insight into neural stem cell differentiation.
Until 7 DIV, all the cells displayed a strong SmartFlareTM fluorescent signal indicative of CD133 and OCT4 mRNA expression, as single dots encompassing both the cytoplasmic domain and the related processes. Upon 15 DIV, cells showed a marked decrease in the fluorescence, both for CD133 and Oct4 probes. In cells grown for 30 DIV, the CD133 and Oct4 probe uptake was very scant but still consisted of single dot-like elements, representative of a downregulation of the same genes.
Our findings propose the SmartFlareTM technology as a reliable and straightforward tool in the context of a qualitative expression analysis applied to a broad panel of mRNAs in single living stem cells.
The NanoFlare technology, such as SmartFlareTM, could enhance the scenario of biomedical applications in the field of marker identification mirroring both normal and pathological conditions, with the advantage of ensuring the physiological integrity of cellular systems.