Amongst different living organisms studied as potential candidates for the green synthesis of copper nanoparticles, algal biomass is presented as a novel and easy-to-handle method. However, the role of specific biomolecules and their contribution as reductant and capping agents has not yet been described. This contribution reports a green synthesis method to obtain copper oxide nanoparticles (CuO-NPs) using separated protein fractions from an aqueous extract of brown algae Macrocystis pyrifera through size exclusion chromatography (HPLC-SEC). Proteins were detected by a UV/VIS diode array, time-based fraction collection was carried out, and each collected fraction was used to evaluate the synthesis of CuO-NPs. The characterization of CuO-NPs was evaluated by Dynamic Light Scattering (DLS), Z-potential, Fourier Transform Infrared (FTIR), Transmission Electron Microscope (TEM) equipped with Energy Dispersive X-ray Spectroscopy (EDS) detector. Low Molecular Weight (LMW) and High Molecular Weight (HMW) protein fractions were able to synthesize spherical CuO-NPs. TEM images showed that the metallic core present in the observed samples ranged from 2 to 50 nm in diameter, with spherical nanostructures present in all containing protein samples. FTIR measurements showed functional groups from proteins having a pivotal role in the reduction and stabilization of the nanoparticles. The highly negative zeta potential average values from obtained nanoparticles suggest high stability, expanding the range of possible applications. This facile and novel protein-assisted method for the green synthesis of CuO-NPs may also provide a suitable tool to synthesize other nanoparticles that have different application areas.

Colorectal cancer (CRC) is the third most prevalent cancer with the second highest mortality rate worldwide. CRC is a heterogenous disease with multiple risk factors associated, including obesity, smoking, and use of alcohol. Of total CRC cases, 60% are diagnosed in late stages, where survival can drop to about 10%. CRC screening programs are based primarily on colonoscopy, yet this approach is invasive and has low patient adherence. Therefore, there is a strong incentive for developing molecular-based methods that are minimally invasive and have higher patient adherence. Recent reports have highlighted the importance of extracellular vesicles (EVs), specifically exosomes, as intercellular communication vehicles with a broad cargo, including micro-RNAs (miRNAs). These have been syndicated as robust candidates for diagnosis, primarily for their known activities in cancer cells, including immunoevasion, tumor progression, and angiogenesis, whereas miRNAs are dysregulated by cancer cells and delivered by cancer-derived exosomes (CEx). Quantitative polymerase chain reaction (qPCR) has shown good results detecting specific cancer-derived exosome micro-RNAs (CEx-miRNAs) associated with CRC, but qPCR also has several challenges, including portability and sensitivity/specificity issues regarding experiment design and sample quality. CRISPR/Cas-based platforms have been presented as cost-effective, ultrasensitive, specific, and robust clinical detection tools in the presence of potential inhibitors and capable of delivering quantitative and qualitative real-time data for enhanced decision-making to healthcare teams. Thereby, CRISPR/Cas13-based technologies have become a potential strategy for early CRC diagnosis detecting CEx-miRNAs. Moreover, CRISPR/Cas13-based platforms’ ease of use, scalability, and portability also showcase them as a potential point-of-care (POC) technology for CRC early diagnosis. This study presents two potential CRISPR/Cas13-based methodologies with a proposed panel consisting of four CEx-miRNAs, including miR-126, miR-1290, miR-23a, and miR-940, to streamline novel applications which may deliver a potential early diagnosis and prognosis of CRC.

Almost all of Earth's oceans are now impacted by multiple anthropogenic stressors, including the spread of nonindigenous species, harmful algal blooms, and pathogens. Early detection is critical to manage these stressors effectively and to protect marine systems and the ecosystem services they provide. Molecular tools have emerged as a promising solution for marine biomonitoring. One of the latest advancements involves utilizing CRISPR-Cas technology to build programmable, rapid, ultrasensitive, and specific diagnostics. CRISPR-based diagnostics (CRISPR-Dx) has the potential to allow robust, reliable, and cost-effective biomonitoring in near real time. However, several challenges must be overcome before CRISPR-Dx can be established as a mainstream tool for marine biomonitoring. A critical unmet challenge is the need to design, optimize, and experimentally validate CRISPR-Dx assays. Artificial intelligence has recently been presented as a potential approach to tackle this challenge. This perspective synthesizes recent advances in CRISPR-Dx and machine learning modeling approaches, showcasing CRISPR-Dx potential to progress as a rising molecular tool candidate for marine biomonitoring applications.

Marine sponges have recently emerged as efficient natural environmental DNA (eDNA) samplers. The ability of sponges to accumulate eDNA provides an exciting opportunity to reconstruct contemporary communities and ecosystems with high temporal and spatial precision. However, the use of historical eDNA, trapped within the vast number of specimens stored in scientific collections, opens up the opportunity to begin to reconstruct the communities and ecosystems of the past. Here, we define the term ‘heDNA’ to denote the historical environmental DNA that can be obtained from the recent past with high spatial and temporal accuracy. Using a variety of Antarctic sponge specimens stored in an extensive marine invertebrate collection, we were able to recover information on Antarctic fish biodiversity from specimens up to 20 years old. We successfully recovered 64 fish heDNA signals from 27 sponge specimens. Alpha diversity measures did not differ among preservation methods, but sponges stored frozen had a significantly different fish community composition compared to those stored dry or in ethanol. Our results show that we were consistently and reliably able to extract the heDNA trapped within marine sponge specimens, thereby enabling the reconstruction and investigation of communities and ecosystems of the recent past with a spatial and temporal resolution previously unattainable. Future research into heDNA extraction from other preservation methods, as well as the impact of specimen age and collection method, will strengthen and expand the opportunities for this novel resource to access new knowledge on ecological change during the last century.

Environmental biosecurity challenges are intensifying as climate change and human activities accelerate the spread of invasive species, disrupting ecosystem composition, function, and essential services. Environmental DNA (eDNA) has transformed traditional biosurveillance by detecting trace DNA fragments left by organisms in their surroundings, primarily by applying quantitative polymerase chain reaction (qPCR) methods. However, qPCR presents challenges, including limited portability, reliance on precise thermal cycling, and susceptibility to inhibitors. To address these challenges and enable field‐deployable monitoring, isothermal amplification techniques such as recombinase polymerase amplification (RPA) paired with clustered regularly interspaced short palindromic repeats and associated proteins (CRISPR‐Cas) have been proposed as promising alternatives. CRISPR‐Cas technology also presents challenges, including searching and optimizing a guide RNA (gRNA) that is highly sensitive and has no off‐target interactions for use as an effective environmental biosurveillance tool. We present here the development of SENTINEL (Smart Environmental Nucleic‐acid Tracking using Inference from Neural‐networks for Early‐warning Localization) that harnesses the programmability, specificity and sensitivity of a one‐pot RPA‐CRISPR‐Cas13a reaction by integrating an accessible and pre‐trained neural network to accelerate assay design for rapid deployment. We challenged SENTINEL with waterborne eDNA from two marine sites invaded by species not native to New Zealand as proof‐of‐concept fluorescence‐based tests: Sabella spallanzanii (Mediterranean fanworm) and Undaria pinnatifida (Wakame). Off‐target effects were explored by challenging the SENTINEL assays on gDNA from a suite of co‐occurring species. SENTINEL presented a robust, streamlined method incorporating the trained neural network, achieving a sensitivity down to 10 attomolar using recombinant DNA and down to ~0.34 copies/μL for eDNA samples with 1 h, costing 3.5 USD per sample. There was a 100% agreement between SENTINEL results and qPCR‐based analysis of the eDNA samples. SENTINEL displayed no off‐target activity when challenged against 23 gDNA samples from co‐occurring species. Thus, our study showcases SENTINEL's potential as a robust platform for eDNA screening applications.