Chemically Conjugated Branched Staples for Super-DNA Origami.

DNA origami, comprising a long folded DNA scaffold and hundreds of linear DNA staple strands, has been developed to construct various sophisticated structures, smart devices, and drug delivery systems. However, the size and diversity of DNA origami are usually constrained by the length of DNA scaffolds themselves. Herein, we report a new paradigm of scaling up DNA origami assembly by introducing a novel branched staple concept. Owing to their covalent characteristics, the chemically conjugated branched DNA staples we describe here can be directly added to a typical DNA origami assembly system to obtain super-DNA origami with a predefined number of origami tiles in one pot. Compared with the traditional two-step coassembly system (yields <10%), a much greater yield (>80%) was achieved using this one-pot strategy. The diverse superhybrid DNA origami with the combination of different origami tiles can be also efficiently obtained by the hybrid branched staples. Furthermore, the branched staples can be successfully employed as the effective molecular glues to stabilize micrometer-scale, super-DNA origami arrays (e.g., 10 × 10 array of square origami) in high yields, paving the way to bridge the nanoscale precision of DNA origami with the micrometer-scale device engineering. This rationally developed assembly strategy for super-DNA origami based on chemically conjugated branched staples presents a new avenue for the development of multifunctional DNA origami-based materials.

Genetically Encoded DNA Origami for Gene Therapy In Vivo.

DNA origami has played an important role in various biomedical applications, including biosensing, bioimaging, and drug delivery. However, the function of the long DNA scaffold involved in DNA origami has yet to be fully exploited. Herein, we report a general strategy for the construction of a genetically encoded DNA origami by employing two complementary DNA strands of a functional gene as the DNA scaffold for gene therapy. In our design, the complementary sense and antisense strands can be directly folded into two DNA origami monomers by their corresponding staple strands. After hybridization, the assembled genetically encoded DNA origami with precisely organized lipids on the surface can function as the template for lipid growth. The lipid-coated and genetically encoded DNA origami can efficiently penetrate the cell membrane for successful gene expression. After decoration with the tumor-targeting group, the antitumor gene (p53) encoded DNA origami can elicit a pronounced upregulation of the p53 protein in tumor cells to achieve efficient tumor therapy. The targeting group-modified, lipid-coated, and genetically encoded DNA origami has mimicked the functions of cell surface ligands, cell membrane, and nucleus for communication, protection, and gene expression, respectively. This rationally developed combination of folding and coating strategies for genetically encoded DNA origami presents a new avenue for the development of gene therapy.

A DNA Origami-Based Gene Editing System for Efficient Gene Therapy in Vivo.

DNA nanostructures have played an important role in the development of novel drug delivery systems. Herein, we report a DNA origami-based CRISPR/Cas9 gene editing system for efficient gene therapy in vivo. In our design, a PAM-rich region precisely organized on the surface of DNA origami can easily recruit and load sgRNA/Cas9 complex by PAM-guided assembly and pre-designed DNA/RNA hybridization. After loading the sgRNA/Cas9 complex, the DNA origami can be further rolled up by the locking strands with a disulfide bond. With the incorporation of DNA aptamer and influenza hemagglutinin (HA) peptide, the cargo-loaded DNA origami can realize the targeted delivery and effective endosomal escape. After reduction by GSH, the opened DNA origami can release the sgRNA/Cas9 complex by RNase H cleavage to achieve a pronounced gene editing of a tumor-associated gene for gene therapy in vivo. This rationally developed DNA origami-based gene editing system presents a new avenue for the development of gene therapy.

Hierarchical Assembly of Super-DNA Origami Based on a Flexible and Covalent-Bound Branched DNA Structure.

DNA origami technique provides a programmable way to construct nanostructures with arbitrary shapes. The dimension of assembled DNA origami, however, is usually limited by the length of the scaffold strand. Herein, we report a general strategy to efficiently organize multiple DNA origami tiles to form super-DNA origami using a flexible and covalent-bound branched DNA structure. In our design, the branched DNA structures (Bn: with a certain number of 2-6 branches) are synthesized by a copper-free click reaction. Equilateral triangular DNA origamis with different numbers of capture strands (Tn: T1, T2, and T3) are constructed as the coassembly tiles. After hybridization with the branched DNA structures, the super-DNA origami (up to 13 tiles) can be efficiently ordered in the predesigned patterns. Compared with traditional DNA junctions (Jn: J2-J6, as control groups) assembled by base pairing between several DNA strands, a higher yield and more compact structures are obtained using our strategy. The highly ordered and discrete DNA origamis can further precisely organize gold nanoparticles into different patterns. This rationally developed DNA origami ordering strategy based on the flexible and covalent-bound branched DNA structure presents a new avenue for the construction of sophisticated DNA architectures with larger molecular weights.

Branched Antisense and siRNA Co-Assembled Nanoplatform for Combined Gene Silencing and Tumor Therapy.

Chemically modified DNA has been widely developed to fabricate various nucleic acid nanostructures for biomedical applications. Herein, we report a facile strategy for construction of branched antisense DNA and small interfering RNA (siRNA) co-assembled nanoplatform for combined gene silencing in vitro and in vivo. In our design, the branched antisense can efficiently capture siRNA with 3' overhangs through DNA-RNA hybridization. After being equipped with an active targeting group and an endosomal escape peptide by host-guest interaction, the tailored nucleic acid nanostructure functions efficiently as both delivery carrier and therapeutic cargo, which is released by endogenous RNase H digestion. The multifunctional nucleic acid nanosystem elicits an efficient inhibition of tumor growth based on the combined gene silencing of the tumor-associated gene polo-like kinase 1 (PLK1). This biocompatible nucleic acid nanoplatform presents a new strategy for the development of gene therapy.

A Self-Assembled Platform Based on Branched DNA for sgRNA/Cas9/Antisense Delivery.

Precisely assembled DNA nanostructures are promising candidates for the delivery of biomolecule-based therapeutics. Herein, we introduce a facile strategy for the construction of a branched DNA-based nanoplatform for codelivery of gene editing (sgRNA/Cas9, targeting DNA in the nucleus) and gene silencing (antisense, targeting mRNA in the cytoplasm) components for synergistic tumor therapy in vitro and in vivo. In our design, the branched DNA structure can efficiently load a sgRNA/Cas9/antisense complex targeting a tumor-associated gene, PLK1, through DNA self-assembly. With the incorporation of an active targeting aptamer and an endosomal escape peptide by host-guest interaction, the biocompatible DNA nanoplatform demonstrates efficient inhibition of tumor growth without apparent systemic toxicity. This multifunctional DNA nanocarrier provides a new strategy for the development of gene therapeutics.

Enantioselective Cascade Reaction for Synthesis of Quinolinones through Synergistic Catalysis Using Cu-Pybox and Chiral Benzotetramisole as Catalysts.

In contrast to the well-studied asymmetric catalyzed synthesis of tetrahydroquinolines, the asymmetric methodologies toward 3,4-dihydroquinolin-2-ones are quite rare. Herein, the first asymmetric cascade reaction is reported between ethynyl benzoxazinanones and mixed-anhydrides generated from aryl acetic acids and pivaloyl chloride, based on synergistic catalysis. This allowed the formation of attractive 3,4-dihydroquinolin-2-ones bearing two vicinal chiral centers at C3 and C4 in high yields with excellent diastereo- and enantioselectivities. A plausible chiral induction model for this reaction was proposed. The utility of this methodology was exemplified by further elaboration of the cyclization products by removal of the N-protecting groups.

Dry Particulate Nitrate Deposition in China.

A limited number of ground measurements of dry particulate nitrate deposition (NO3-) makes it difficult and challenging to fully know the status of the spatial and temporal variations of dry NO3- depositions over China. This study tries to expand the ground measurements of NO3- concentrations at monitoring sites to a national scale, based on the Ozone Monitoring Instrument (OMI) NO2 columns, NO2 profiles from an atmospheric chemistry transport model (Model for Ozone and Related chemical Tracers, version 4, MOZART-4) and monitor-based sources, and then estimates the NO3- depositions on a regional scale based on an inferred model. The ground NO2 concentrations were first derived from NO2 columns and the NO2 profiles, and then the ground NO3- concentrations were derived from the ground NO2 concentrations and the relationship between NO2 and NO3- based on Chinese Nationwide Nitrogen Deposition Monitoring Network (NNDMN). This estimated dry NO3- depositions over China will be helpful in determining the magnitude and pollution status in regions without ground measurements, supporting the construction plan of environmental monitoring in future.

Amide-Phosphonium Salt as Bifunctional Phase Transfer Catalyst for Asymmetric 1,6-Addition of Malonate Esters to para-Quinone Methides.

Asymmetric 1,6-addition of malonates to para-quinone methides has been developed by using amide-phosphonium salts derived from easily available chiral α-amino acids as bifunctional phase transfer catalysts. Stabilized para-quinone methides with various substituents on the phenyl ring were reacted with diphenyl malonates to give functionalized diaryl methines in excellent yields and high to excellent ee's. Furthermore, to show the utility of this methodology, a gram scale synthesis of an 1,6-addition adduct and its further elaboration into the key intermediate for synthesis of GPR40 agonists were also described.

A database of atmospheric inorganic nitrogen deposition fluxes in China from satellite monitoring.

Over the past century, atmospheric inorganic nitrogen (IN) deposition to terrestrial ecosystems has significantly increased and caused various environmental issues. China has been one of the hotspot regions for IN deposition, yet limited data exist regarding IN deposition fluxes in China at the regional scale. In this study, based on NO2 and NH3 columns acquired by satellite sensors, coupled with atmospheric chemical transport model (CTM), mixed-effects model and site observations, we constructed regional-scale IN dry and wet deposition models respectively, and finally proposed a spatially explicit database of IN deposition fluxes in China. The database includes the dry, wet and total deposition fluxes in China during 2011-2020, and the data are presented in raster form with a resolution of 0.25° × 0.25°. Overall, the database is of great importance for monitoring and simulating the trends of IN deposition over a long time series in China.

A covalently conjugated branched DNA aptamer cluster-based nanoplatform for efficiently targeted drug delivery.

Targeted delivery of therapeutic drugs is essential for precise treatment of various diseases to reduce possible serious side-effects. A screened DNA aptamer has been widely developed for active targeting delivery. Herein, we report a facile strategy for the construction of a branched DNA aptamer cluster-based nanoplatform for efficiently targeted drug delivery. In our design, the terminal-modified DNA aptamer can be covalently conjugated to form a branched aptamer cluster by click reaction easily. The branched aptamer cluster-modified DNA tetrahedron (TET) demonstrates highly targeted cellular uptake with the modification of only one site. After loading the chemotherapeutic drug (doxorubicin, DOX), the DNA aptamer cluster-based nanoplatform elicits a remarkable and selective inhibition of tumor cell proliferation by much-enhanced targeted delivery. This covalently conjugated branched DNA aptamer cluster-based nanoplatform provides a new strategy for the development of targeted drug delivery.

Chemically modified DNA nanostructures for drug delivery.

Based on predictable, complementary base pairing, DNA can be artificially pre-designed into versatile DNA nanostructures of well-defined shapes and sizes. With excellent addressability and biocompatibility, DNA nanostructures have been widely employed in biomedical research, such as bio-sensing, bio-imaging, and drug delivery. With the development of the chemical biology of nucleic acid, chemically modified nucleic acids are also gradually developed to construct multifunctional DNA nanostructures. In this review, we summarize the recent progress in the construction and functionalization of chemically modified DNA nanostructures. Their applications in the delivery of chemotherapeutic drugs and nucleic acid drugs are highlighted. Furthermore, the remaining challenges and future prospects in drug delivery by chemically modified DNA nanostructures are discussed.

Assessing the Effects of Human Activities on Terrestrial Net Primary Productivity of Grasslands in Typical Ecologically Fragile Areas.

Global enhanced human activities have deeply influenced grassland ecosystems. Quantifying the impact of human activities on grasslands is crucial to understanding the grassland dynamic change mechanism, such as grassland degradation, and to establishing ecosystem protection measures. In this study, potential net primary productivity (PNPP), actual NPP (ANPP), and the forage harvest NPP (HNPP) were employed to establish the human activities index (HAI) to reveal the spatiotemporal changes of the effects of human activities on grassland ecosystems in eastern Inner Mongolia from 2000 to 2017, and to further explore the relationship between human activities and grassland degradation. The results showed that the total average PNPP, ANPP, and HNPP of grasslands in eastern Inner Mongolia were 187.2 Tg C yr-1, 152.3 Tg C yr-1, and 8.9 Tg C yr-1, respectively, during the period of 2000 to 2017. The HAI exhibited a clear decreasing trend during the study period, with annual mean values ranging from 0.75 to 0.47, which indicates that the NPP loss induced by human activities is weakening, and this trend is dominated by the difference between potential NPP and actual NPP. About 42.4% of the study area was non-degraded grassland, and the declining grassland degradation index (GDI) indicated that the degradation grade in eastern Inner Mongolia improved from moderate to light degradation. A positive relationship was found between HAI and GDI. This relationship was more significant in Xilingol League, which is a typical ecologically fragile area, than that in Xing'an League and Hulunbuir City.

An Aptamer-Modified DNA Tetrahedron-Based Nanogel for Combined Chemo/Gene Therapy of Multidrug-Resistant Tumors.

DNA-based nanogels have attracted much attention in the biomedical research field. Herein, we report a universal strategy for the fabrication of an aptamer-modified DNA tetrahedron (TET)-based nanogel for combined chemo/gene therapy of multidrug-resistant tumors. In our design, terminal extended antisense oligonucleotides (ASOs) are employed as the linker to co-assemble with two kinds of three-vertex extended TETs for the efficient construction of the DNA-based nanogel. With the incorporation of an active cell-targeting group (aptamer in one vertex of TET) and a controlled-release element (disulfide bridges in the terminals of ASOs), the functional DNA-based nanogel can achieve targeted cellular internalization and stimuli-responsive release of embedded ASOs. After loading with the chemodrug (doxorubicin (DOX), an intercalator of double-stranded DNA), the multifunctional DOX/Nanogel elicits efficient chemo/gene therapy of human MCF-7 breast tumor cells with DOX resistance (MCF-7R). This aptamer-modified DNA tetrahedron-based nanogel provides another strategy for intelligent drug delivery and combined tumor therapy.

A Nucleic Acid/Gold Nanorod-Based Nanoplatform for Targeted Gene Editing and Combined Tumor Therapy.

The CRISPR/Cas9 gene-editing system has become a promising strategy for tumor therapy with its powerful oncogene-editing ability. However, the efficient delivery of sgRNA/Cas9 complex into target tumor cells remains a challenge. Herein, we report a facile strategy for the construction of an sgRNA/Cas9 complex co-assembled nanoplatform for targeted gene editing and combined tumor therapy. In our design, the TAT peptide and thiolated DNA linker functionalized gold nanorod can efficiently load the sgRNA/Cas9 complex through the hybridization between the 3' overhang of sgRNA and the DNA linker. Due to the integration of an active cell targeting group (aptamer) and nuclear targeting peptide (TAT), the multifunctional nanoplatform can elicit the targeted cellular internalization and efficient nuclear targeting transportation to realize endogenous RNase H activated gene editing of the tumor-associated gene polo-like kinase 1 (PLK1). With mild photothermal treatment, this sgRNA/Cas9 complex loaded nanoplatform achieved efficient inhibition of tumor cell proliferation. This multifunctional nanocarrier provides a new strategy for the development of combined tumor therapy.

Efficient construction of a stable linear gene based on a TNA loop modified primer pair for gene delivery.

A terminal-closed linear gene with strong exonuclease resistance and serum stability was successfully constructed by polymerase chain reaction (PCR) with an α-l-threose nucleic acid (TNA) loop modified primer pair, which can be used as an efficient gene expression system in eukaryotic cells for gene delivery.

Ammonia volatilization as the major nitrogen loss pathway in dryland agro-ecosystems.

The losses of excessive reactive nitrogen (N) from agricultural production pose detrimental impacts on water, air and land. However, N budgets of agroecosystems are still poorly quantified, presenting a barrier to understand the N turnover in agriculture. Agricultural ammonia (NH3) volatilization has been recognized as a crucial contribution to the pollution of fine particulate matters over China through reacting with acid gases. Building on these challenges, the first national-scale model analysis was constructed on the N budgets to gain an overall insight into the current status of N flows in Chinese dryland systems towards sustainable N management. Total inputs of soil N in Chinese dryland soils were estimated at 121 kg N ha-1 in 2010, considering all pathways including N manure, fertilizer, atmospheric deposition and litter from crop residues. Atmospheric N deposition accounted for 25% of N fertilizer plus N manure in Chinese dryland soils, suggesting that N deposition could not be ignored when estimating total N inputs to Chinese dryland soils. The highest ratio of NH3 volatilization to total N outputs was found at 43 kg N ha-1 (∼21%) in Northern China, followed by 41 kg N ha-1 (∼20%) in Sichuan Basin and 25 kg N ha-1 (∼26%) in Northeastern China. The modeling results indicated that, if a 20% decrease in N fertilizer plus N manure was achieved, it would lead to a 24% (7-49%) reduction in NH3 volatilization. Substantial reductions of NH3 volatilization would also be achieved by making an improvement in changing management practices (controlled release fertilizer and full irrigation). The results would give an overall insight into N budgets in Chinese dryland soils. The constructed N budgets assisted with understanding agricultural N flows and NH3 pollution, and evaluated the impacts of human activities on N cycle towards a precise way to regulate agricultural management.

Challenges for Global Sustainable Nitrogen Management in Agricultural Systems.

Nitrogen (N) losses from agricultural production contribute to detrimental impacts on water, soil, air, and human health. However, it is still lacking in evaluating global N budgets in agricultural systems. Hence, we conducted a global analysis on the current status of the N flows in the agricultural systems, explored the possible mitigation measures and challenges, and investigated the existing regulations on controlling N pollution. Globally, agricultural soils received a total of 73 kg of N ha-1 year-1 on average, including N fertilizer plus manure (61%), atmospheric N deposition (10%), and N litters and fixation (29%). The estimated global NH3 loss to total N inputs was 17%, which led to a loss of $15 billion year-1. The N use efficiency (NUE) in Eastern China (33%) was much lower than that in the Eastern United States (65%) or Western Europe (61%), leaving much room to enhance the NUE to increase agricultural food production. Meanwhile, higher NH3 losses from N fertilizers and manure were found in Eastern China (22%) than the Eastern United States (17%) and Western Europe (17%). We highlight the urgency to improve the NUE and decrease NH3 loss with lower environmental consequences. Our results showed high potentials to mitigate NH3 volatilization and enhance the NUE by various measures, such as substituting manure N for chemical fertilizer N, applying controlled release fertilizers, and urease inhibitors. These measures should be implemented in combination with the transfer of knowledge to farmers with new technologies and increasing the farm size to enhance the efficiency of agricultural production in the future.

Multifunctional DNA Origami Nanoplatforms for Drug Delivery.

DNA nanotechnology has been employed in the construction of self-assembled nano-biomaterials with uniform size and shape for various biological applications, such as bioimaging, diagnosis, or therapeutics. Herein, recent successful efforts to utilize multifunctional DNA origami nanoplatforms as drug-delivery vehicles are reviewed. Diagnostic and therapeutic strategies based on gold nanorods, chemotherapeutic drugs, cytosine-phosphate-guanine, functional proteins, gene drugs, and their combinations for optoacoustic imaging, photothermal therapy, chemotherapy, immunological therapy, gene therapy, and coagulation-based therapy are summarized. The challenges and opportunities for DNA-based nanocarriers for biological applications are also discussed.

Vegetation structural change since 1981 significantly enhanced the terrestrial carbon sink.

Satellite observations show that leaf area index (LAI) has increased globally since 1981, but the impact of this vegetation structural change on the global terrestrial carbon cycle has not been systematically evaluated. Through process-based diagnostic ecosystem modeling, we find that the increase in LAI alone was responsible for 12.4% of the accumulated terrestrial carbon sink (95 ± 5 Pg C) from 1981 to 2016, whereas other drivers of CO2 fertilization, nitrogen deposition, and climate change (temperature, radiation, and precipitation) contributed to 47.0%, 1.1%, and -28.6% of the sink, respectively. The legacy effects of past changes in these drivers prior to 1981 are responsible for the remaining 65.5% of the accumulated sink from 1981 to 2016. These results refine the attribution of the land sink to the various drivers and would help constrain prognostic models that often have large uncertainties in simulating changes in vegetation and their impacts on the global carbon cycle.