Topological barrier to Cas12a activation by circular DNA nanostructures facilitates autocatalysis and transforms DNA/RNA sensing.

Control of CRISPR/Cas12a trans-cleavage is crucial for biosensor development. Here, we show that small circular DNA nanostructures which partially match guide RNA sequences only minimally activate Cas12a ribonucleoproteins. However, linearizing these structures restores activation. Building on this finding, an Autocatalytic Cas12a Circular DNA Amplification Reaction (AutoCAR) system is established which allows a single nucleic acid target to activate multiple ribonucleoproteins, and greatly increases the achievable reporter cleavage rates per target. A rate-equation-based model explains the observed near-exponential rate trends. Autocatalysis is also sustained with DNA nanostructures modified with fluorophore-quencher pairs achieving 1 aM level (<1 copy/µL) DNA detection (106 times improvement), without additional amplification, within 15 min, at room temperature. The detection range is tuneable, spanning 3 to 11 orders of magnitude. We demonstrate 1 aM level detection of SNP mutations in circulating tumor DNA from blood plasma, genomic DNA (H. Pylori) and RNA (SARS-CoV-2) without reverse transcription as well as colorimetric lateral flow tests of cancer mutations with ~100 aM sensitivity.

Improved effectiveness of X-PDT against human triple-negative breast cancer cells through the use of liposomes co-loaded with protoporphyrin IX and perfluorooctyl bromide.

In this study, we utilized X-ray-induced photodynamic therapy (X-PDT) against triple-negative breast cancer (TNBC) cells. To achieve this, we developed a liposome delivery system that co-loaded protoporphyrin IX (PPIX) and perfluorooctyl bromide (PFOB) in a rational manner. Low-dose X-ray at 2 Gy was employed to activate PPIX for the generation of reactive oxygen species (ROS), and the co-loading of PFOB provided additional oxygen to enhance ROS production. The resulting highly toxic ROS effectively induced cell death in TNBC. In vitro X-PDT effects, including intracellular ROS generation, cell viability, and apoptosis/necrosis assays in TNBC cells, were thoroughly investigated. Our results indicate that the nanocarriers effectively induced X-PDT effects with very low-dose radiation, making it feasible to damage cancer cells. This suggests the potential for the effective utilization of X-PDT in treating hypoxic cancers, including TNBC, with only a fraction of conventional radiotherapy.

Gallium Nanodroplets are Anti-Inflammatory without Interfering with Iron Homeostasis.

Gallium (Ga) compounds, as the source of Ga ions (Ga3+), have been historically used as anti-inflammatories. Currently, the widely accepted mechanisms of the anti-inflammatory effects for Ga3+ are rationalized on the basis of their similarities to ferric ions (Fe3+), which permits Ga3+ to bind with Fe-binding proteins and subsequently disturbs the Fe homeostasis in the immune cells. Here in contrast to the classic views, our study presents the mechanisms of Ga as anti-inflammatory by delivering Ga nanodroplets (GNDs) into lipopolysaccharide-induced macrophages and exploring the processes. The GNDs show a selective inhibition of nitric oxide (NO) production without affecting the accumulation of pro-inflammatory mediators. This is explained by GNDs disrupting the synthesis of inducible NO synthase in the activated macrophages by upregulating the levels of eIF2α phosphorylation, without interfering with the Fe homeostasis. The Fe3+ transferrin receptor-independent endocytosis of GNDs by the cells prompts a fundamentally different mechanism as anti-inflammatories in comparison to that imparted by Ga3+. This study reveals the fundamental molecular basis of GND-macrophage interactions, which may provide additional avenues for the use of Ga for anti-inflammatory and future biomedical and pharmaceutical applications.

Recent advances in liposome formulations for breast cancer therapeutics.

Among many nanoparticle-based delivery platforms, liposomes have been particularly successful with many formulations passed into clinical applications. They are well-established and effective gene and/or drug delivery systems, widely used in cancer therapy including breast cancer. In this review we discuss liposome design with the targeting feature and triggering functions. We also summarise the recent progress (since 2014) in liposome-based therapeutics for breast cancer including chemotherapy and gene therapy. We finally identify some challenges on the liposome technology development for the future clinical translation.

Spatial and Temporal Control of CRISPR-Cas9-Mediated Gene Editing Delivered via a Light-Triggered Liposome System.

The CRISPR-Cas9 and related systems offer a unique genome-editing tool allowing facile and efficient introduction of heritable and locus-specific sequence modifications in the genome. Despite its molecular precision, temporal and spatial control of gene editing with the CRISPR-Cas9 system is very limited. We developed a light-sensitive liposome delivery system that offers a high degree of spatial and temporal control of gene editing with the CRISPR-Cas9 system. We demonstrated its efficient protein release by respectively assessing the targeted knockout of the eGFP gene in human HEK293/GFP cells and the TNFAIP3 gene in TNFα-induced HEK293 cells. We further validated our results at a single-cell resolution using an in vivo eGFP reporter system in zebrafish (77% knockout). These findings indicate that light-triggered liposomes may have new options for precise control of CRISPR-Cas9 release and editing.

A Sensing Peak Identification Method for Fiber Extrinsic Fabry⁻Perot Interferometric Refractive Index Sensing.

A novel sensing peak identification method for high accuracy refractive index (RI) sensing is proposed. The implementation takes the intensity of interference maximum as the characteristic to distinguish interference peaks, tracking the sensing peak continually during a RI changes, with high measurement accuracy and simple computation. To verify the effect of the method, the extrinsic Fabry⁻Perot interferometer (EFPI) sensor has been fabricated using the large lateral offset splicing technique. In the RI range from 1.346 to 1.388, the measurement range of the EFPI with the proposed method reaches at least 6 times larger than that of EFPI with the wavelength tracking method and the largest measurement error is -4.47 × 10-4. The EFPI refractive index (RI) sensor identified the sensing peak is believed to play an important role in RI, concentration and density sensing, etc., for superior performance.

[Analysis of Theory and Performance of Multi Layer Graphene Nanoribbons Photodetector].

Multilayer graphene, with wide absorption spectrum and unique photoelectric properties, is an ideal material to make the next generation of photoelectric detector. Taking graphene interband tunneling theory as the foundation, a photoelectric detector model with the structure of multilayer graphene nanoribbons was proposed. Nanoribbons which contacted with source and drain electrode at the end were sandwiched between the semiconductor substrate and the top and back gate. Using this model, a photoelectric conversion mechanism of multilayer graphene nanoribbon detector was established. It discussed the working principle of the detector at different top gate voltage, studied the relationship between the source-drain current and the incident light energy, researched the influence of the bias voltage, the length of depletion and the values of band gap on the dark current, and analyzed the change of detector responsibility and detectivity with the incident light energy under the different parameters. The results show that, the responsibility of detector increases with the layers of nanoribbons, and are affected by the band gap, the length of depletion and the bias voltage. The maximum responsibility up to 10(3) A·W(-1); By limiting on the top gate voltage, the band gap and other variables can control the dark current of system and increase the detectivity, the detectivity up to a maximum value of 10(9) cm Hz(1/2)·W(-1). The structure of multilayer graphene nanoribbons can enhance the absorption of the incident light, improve the sensitivity of the detector and the detection capability of weak light, and realize the detection from THz to far infrared wavelength of incident light. The detection performance is far better than that of many quantum structures and narrow-band semiconductor structure of photoelectric detector.