Functional Titanium Carbide MXenes-Loaded Entropy-Driven RNA Explorer for Long Noncoding RNA PCA3 Imaging in Live Cells

Abstract

The visualization of the long noncoding RNA of prostate cancer gene 3 (lncRNA PCA3), a specific biomarker for androgen receptor-positive prostate cancer, in living cells not only directly reflects the gene expression and localization but also offers better insight into its roles in the pathological processes. Here, we loaded an entropy-driven RNA explorer (EDRE) on the TAT peptide-functionalized titanium carbide MXenes (Ti3C2–TAT) for the imaging of nuclear lncRNA PCA3 in live cells. The EDRE was condensed on the Ti3C2–TAT (Ti3C2–TAT@EDRE) by electrostatic interaction. Ti3C2–TAT@EDRE enables the entering of cells and release of TAT peptides and EDRE in the cytoplasm by the glutathione (GSH)-triggered cleavage of the disulfide bonds in Ti3C2–TAT. The released EDRE is delivered into the nucleus by the nucleus-targeted guidance of TAT peptides, and initiated by the target lncRNA PCA3, subsequently leading to the continuous accumulation of fluorescence signals. Consequently, fluorescence analysis of lncRNA PCA3 at low-picomolar concentrations in vitro as well as sensitive live cell imaging of lncRNA PCA3 in the nucleus of androgen receptor-positive LNCaP prostate cancer cells were achieved, providing a versatile strategy for the monitoring of nucleic acid biomarkers in the nucleus of living cells.

Mismatched fluorescent probes with an enhanced strand displacement reaction rate for intracellular long noncoding RNA imaging

“…The sensitivity obtained by using the mismatched fluorescent probe 5 is much higher than that obtained by using the matched fluorescent probe 6 (9.8 nM) (Fig. S6, ESI†) and is comparable to those obtained by nonenzymatic isothermal amplifiers for the lncRNA assay (2.6 pM), 17 and accelerated the DNA cascade amplification reaction for the mRNA assay (10 pM). 7 c We further used the nontarget lncRNA metastasis associated lung adenocarcinoma transcript-1 (MALAT1) and nontarget miRNAs ( i.e.…”

Integrating CRISPR-Cas12a with a DNA circuit as a generic sensing platform for amplified detection of microRNA

“…Many studies have revealed that miR-21 plays a pivotal role in numerous biological processes by regulating the post-transcription of gene expression. [44][45][46] The aberrant level of miR-21 is considered to be associated with the etiology, progression, and prognosis of many tumour types, including brain, liver, prostate, and lung cancers. [47][48][49][50] In order to verify whether miR-21 can be converted into DNA duplex outputs by the proposed CHA circuit, miR-21 was incubated with H1 and H2, and the mixture was analysed by gel shi assay.…”

“…[26][27][28][29] Because the functions of the nucleic acid circuits depend solely on the Watson-Crick complementarity, these DNA circuits are also able to work comparably with RNA inputs, including mRNA, long noncoding RNA and miRNA, as with ssDNA inputs. [30][31][32][33][34][35][36] Moreover, the combination of DNA circuits with nucleases has been demonstrated to be effective in the development of highly sensitive assays for miRNA. [37][38][39][40] Encouraged by current progress, we are interested in expanding the application scope of DNAtargeting Cas proteins (e.g., Cas12a) to miRNA sensing using a DNA circuit as the converter.…”

Fuel‐Powered DNA Nanomachines for Biosensing and Cancer Therapy

“…In addition, Wang and co-worker developed a versatile strategy that introduced TAT peptide-modified titanium carbide Mxeners (Ti 3 C 2 -TAT) as a carrier to load an entropy-driven RNA amplifier for the detection of intracellular nuclear IncRNA PCA3. [32] Taking advantage of DNA nanostructures including ease to assembly and precise manipulation of the location and sequence of probes, we constructed a self-powered DNA nanomachine based on orderly DNA self-assembly for miRNA imaging inside living cells (Figure 1D). [33] It is an integrated amplification platform by precisely and orderly assembly entropy-driven substrates, fuel strands and a targeting aptamer on a DNA nanostructure.…”