Biomembrane-inspired lipid nanoparticles enhance CRISPR-Cas9 editing for hemophilia A.

Lipid nanoparticles (LNPs) have substantially advanced RNA-based therapies; however, their use for CRISPR-Cas9 remains limited by sub-optimal endosomal escape, innate immune activation, transient nuclease expression, and restricted tissue specificity. Here, we engineered biomembrane-inspired LNPs containing sphingomyelin and C18-galactosyl ceramide (C18-GalCer) to improve liver-targeted CRISPR delivery. The optimized formulation increased in vivo editing efficiency 2.

A General Strategy to Develop Highly Sensitive FAPα Fluorescent Probes for Invasive Cancer Detection.

Fibroblast activation protein α (FAPα) has been reported to be expressed in many carcinoma cells and associated with tumor growth, invasive, and metastasis. However, due to the lack of appropriate fluorescent probes, detecting FAPα with high sensitivity in tumor and clearly delineating tumor margins remains a challenge. Herein, we developed a general design strategy-introducing a positive charge into the fluorophore skeleton-for the construction of FAPα-activated fluorescent probe with enhanced sensitivity.

Engineering molecular switches of CRISPR/Cas12a for biosensing applications.

Benefiting from high specificity, excellent programmability, and efficient signal amplification capability, the CRISPR/Cas12a system has emerged as a crucial tool in the field of molecular diagnostics. To fully realize the application potential of Cas12a, many molecular switches of CRISPR/Cas12a have been recently developed to convert the activity of Cas12a from an inactive state to an active state in response to external stimuli, which significantly improves the biosensing performance in terms of the sensitivity, selectivity, and accuracy. This review summarizes recent progress in the development and application of Cas12a-based molecular switches, with a focus on their conditional activation mechanisms and biosensing applications.

CRISPR-Based Live Cell Biosensing: Innovations in Signal Labeling Strategies and Delivery Systems.

CRISPR technology provides unprecedented precision for molecular imaging and target detection at the cellular level. However, interested researchers inevitably encounter challenges, including weak labeling signals, low delivery efficiency, and off-target effects when establishing application frameworks. Therefore, discussing signal labeling strategies and delivery systems is crucial to further improving the performance of CRISPR-based live cell biosensing.

A dual-excitation ratiometric NIR-II fluorescent nanoplatform enables high contrast imaging.

Ratiometric fluorescent probes in the second near-infrared window (NIR-II) with a self-calibration function are sought after for reliable imaging of physiological and pathological processes. Nevertheless, current ratiometric NIR-II fluorescent probes usually show severe spectral overlap in the emission channel, resulting in inevitable sacrifice of the emission intensity of the probe and compromised imaging quality in the NIR-II region. To address these challenges, we developed a novel dual-excitation ratiometric NIR-II fluorescence nanoplatform (DERF-NP), in which the intensity ratio of the same full-wavelength emission from 1000 to 1700 nm under two non-overlapping monochrome excitations with distinct responses is conventionally defined as the quantification parameter.

Multimerin-1 modulates macrophage M2 polarization and enhances tumor cell stemness in glioblastoma.

Objectives: Glioblastoma (GBM) is one of the most aggressive brain tumors, with a poor prognosis. Brain tumor stem cells (BTSCs) play a central role in GBM progression and recurrence. This study aimed to identify key BTSC-related genes associated with GBM prognosis and explore their potential biological functions.

Development of NIR-II Ratiometric Fluorescent Nanoprobe for HClO Imaging in Acute Lung Injury.

Efficient and high-contrast bioimaging of acute lung injury (ALI) is crucial for studying disease progression and enabling timely interventional treatments. However, developing bioimaging probes in the second near-infrared (NIR-II) window to assess ALI remains challenging. In particular, ratiometric NIR-II probes with self-calibration functions are rare, hindering reliable detection of information during the ALI process.

Harnessing Porous Coordination Cages for Sonodynamic Therapy: Enhanced Efficacy Through Atomic Precision and Immune Activation.

Sonodynamic therapy (SDT) has emerged as a promising non-invasive approach for immunotherapy. However, its broad applicability is often limited by the inefficiency of sonosensitizers. Here, we introduce a novel series of porous coordination cages (PCCs) specifically engineered to enhance sonodynamic therapeutic performance for the first time.

Rapid sorting and auxiliary evaluation of malignant breast tumors by accurate imaging analysis of metastasis-related biomarker.

Accurate differentiation of benign and malignant breast tumors is paramount for establishing schemes of breast cancer treatment and prognosis. Here we report a near-infrared (NIR) fluorescence probe (YF-1) with the overexpressed cathepsin C (CTSC) in metastatic breast tumors as the detecting substrate. This probe allows accurate identification of malignant tumor tissue specimens among tumor tissue specimens with unknown properties in a blind study.

Ultrabright difuranfluoreno-dithiophen polymers for enhanced afterglow imaging of atherosclerotic plaques.

Cardiovascular diseases, including stroke driven by atherosclerosis, remain a leading global health concern. Current diagnostic imaging modalities such as magnetic resonance imaging fail to characterize oxidative stress within atherosclerotic plaques. Here, we introduce difuranfluoreno-dithiophen-based polymers designed for afterglow imaging, offering ultrabright luminescence, ultralow-power excitation (0.

Constructing Stable and Wavelength-Extended Heptamethine Cyanines via Donor Ectopic Substitution for NIR-IIa/b Bioimaging.

Organic NIR-II dyes, particularly cyanine fluorophores, offer high molar extinction coefficients, biocompatibility, and structural tunability and are popular for noninvasive, high-resolution, and -contrast in vivo imaging. However, achieving stable, long-wavelength, and large Stokes shift NIR-II cyanine suitable for NIR-IIa/IIb bioimaging is still a formidable challenge. Herein, we introduce a novel strategy that extends the emission wavelength by the enhanced Highest occupied molecular orbital (HOMO)-Lowest occupied molecular orbital (LUMO) separation through simple donor ectopic substitution at the terminal structure of NIR-II cyanine.

Functional SELEX and Biomedical Applications of Aptamers: Beyond Molecular Recognition.

Aptamers, developed through SELEX (systematic evolution of ligands by exponential enrichment), are generally short oligonucleotide molecules with remarkable specificity and binding affinity to diverse biological targets. These molecules have shown promise across such fields as biosensing, molecular diagnostics, and bioimaging. However, while conventional aptamer selection strategies predominantly emphasize binding affinity, they overlook the broader spectrum of potential biological functionalities.

Enhancing hemophilia A gene therapy by strategic F8 deletions in AAV vectors.

Hemophilia A, caused by a deficiency in factor VIII (F8), is a promising target for gene therapy. This study aims to enhance the efficacy of adeno-associated virus serotype 8 (AAV8) vectors, specifically those encoding B-domain-deleted F8 (BDDF8), to treat the condition. We focused on improving therapeutic outcomes by strategically deleting amino acids at the furin cleavage site (RHQR), a modification that is crucial for increasing F8 expression and reducing capsid stress during vector packaging.

Harnessing crRNA Transformer for Facile and Specific Nucleic Acid Detection.

CRISPR/Cas systems have emerged as promising tools for nucleic acid detection. However, their practical applications have been limited by poor specificity and the need for additional preprocessing steps. Inspired by the concept of transformers, we found that changing the forms of crRNA with spatial arrangement may endow CRISPR/Cas with an enhanced performance for nucleic acid detection.

Spatial Confinement of a Dual Activatable DNAzyme Sensor in the Cavity of a DNA Nanocage for Logic-Gated Molecular Imaging.

Despite intense interest in design of DNAzyme sensors for molecular detection and imaging in living cells, their intracellular applications are still hampered by limited spatial control and poor bio-stability. Here we present controlled spatial confinement of a rationally designed, microRNA (miRNA)-activatable DNAzyme sensor probe (mDz) within the cavity of DNA nanocage, enabling efficient intracellular delivery with improved bio-stability for AND-gate molecular imaging. The mDz that possesses inactive DNAzyme activity is designed by the introduction of a blocking DNA strand, while miR-21 mediated strand displacement reaction allows for the formation of an intact DNAzyme structure for metal-ion-mediated catalytic reaction.

Microenvironment-confined kinetic elucidation and implementation of a DNA nano-phage with a shielded internal computing layer.

Multiple receptor analysis-based DNA molecular computation has been developed to mitigate the off-target effect caused by nonspecific expression of cell membrane receptors. However, it is quite difficult to involve nanobodies into molecular computation with programmed recognition order because of the "always-on" response mode and the inconvenient molecular programming. Here we propose a spatial segregation-based molecular computing strategy with a shielded internal computing layer termed DNA nano-phage (DNP) to program nanobody into DNA molecular computation and build a series of kinetic models to elucidate the mechanism of microenvironment-confinement.

Activatable Photosensitizer Prodrug for Self-Amplified Immune Therapy Via Pyroptosis.

Immunotherapy is a promising cancer treatment, but its application is hindered by tumors' low immunogenicity and the difficulty of immune cell infiltration. Here, to address above issues and achieve targeted tumor treatment, we designed the first activated small molecule photosensitizer immune-prodrug HDIM based on pyroptosis, and proposed a self-amplified immune therapy strategy (SITS) for enhanced tumor therapy. HDIM can be specifically activated by the tumor hypoxia and then simultaneously initiate immuno-therapy and photodynamic therapy (PDT)-induced pyroptosis with NIR laser irradiation.

Magnetic-susceptibility-dependent ratiometric probes for enhancing quantitative MRI.

In magnetic resonance imaging (MRI), quantitative measurements of analytes are hindered by difficulties in distinguishing the MRI signals of activation of the probe by the analyte from those of the accumulation of the intact probe. Here we show that imaging sensitivity and quantitation can be enhanced by ratiometric MRI probes with a high relaxivity-ratio change (more than 2.5-fold at 7 T) via magnetic-susceptibility-dependent magnetic resonance tuning.

Responsive probes for in vivo magnetic resonance imaging of nitric oxide.

Nitric oxide (NO), a pivotal signalling molecule, plays multifaceted roles in physiological and pathological processes, including cardiovascular and immune functions, neurotransmission and cancer progression. Nevertheless, measuring NO in vivo is challenging due to its transient nature and the complexity of the biological environment. Here we describe NO-responsive magnetic probes made of crosslinked superparamagnetic iron oxide nanoparticles tethered to a NO-sensitive cleavable linker for highly sensitive and selective NO magnetic resonance imaging in vivo.

Development of Dual-Responsive Fluorescent Probe for Drug Screening of Diabetes Cardiomyopathy.

For specific drug research and development, a drug-screening strategy (DSS) plays an indispensable role in the biomedical field. Unfortunately, traditional strategies are complicated and insufficiently accurate due to the widely used single-target screening method. Herein, a simple dual-target-based drug-screening strategy (dt-DSS) is proposed to screen highly effective drugs by fluorescence imaging.

Ultrabright and ultrafast afterglow imaging in vivo via nanoparticles made of trianthracene derivatives.

Low sensitivity, photobleaching, high-power excitation and long acquisition times constrain the utility of afterglow luminescence. Here we report the design and imaging performance of nanoparticles made of electron-rich trianthracene derivatives that, on excitation by room light at ultralow power (58 μW cm), emit afterglow luminescence at ~500 times those of commonly used organic afterglow nanoparticles. The nanoparticles' ultrabright afterglow allowed for deep-tissue imaging (up to 6 cm), for ultrafast afterglow imaging (at short acquisition times down to 0.

Dual mature microRNA-responsive logic biosensing platform based on CRISPR/Cas12a and DNA nanocage.

Mature microRNAs play crucial roles in tumorigenesis and progression. However, their potential as cancer biomarkers is limited by the sequence interference of precursor microRNAs and the occurrence of false positive signals mediated by single microRNAs. Herein, we reported a dual mature microRNA-responsive second-order (YES-AND) logic biosensing platform for accurate cancer diagnosis.

Reactive oxygen species-mediated organic long-persistent luminophores light up biomedicine: from two-component separated nano-systems to integrated uni-luminophores.

Organic luminophores have been widely utilized in cells and fluorescence imaging but face extreme challenges, including a low signal-to-noise ratio (SNR) and even false signals, due to non-negligible background signals derived from real-time excitation lasers. To overcome these challenges, in the last decade, functionalized organic long-persistent luminophores have gained much attention. Such luminophores could not only overcome the biological toxicity of inorganic long-persistent luminescent materials (metabolic toxicity and leakage risk of inorganic heavy metals), but also continue to emit long-persistent luminescence after removing the excitation source, thus effectively improving imaging quality.