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Most patients are at high risk of thrombosis during cancer treatment. However, the major discrepancy in the therapeutic mechanisms and microenvironment between tumors and thrombosis makes it challenging for a panacea to treat cancer while being able to eliminate the risk of thrombosis. Herein, we developed a biomimetic MnOx/Ag

DNA nanotweezers (DTs) are reversible DNA nanodevices that can optionally switch between opened and closed states. Due to their excellent flexibility and high programmability, they have been recognized as a promising platform for constructing a diversity of biosensors and logic gates, as well as a versatile tool for molecular biology studies. In this review, we provide an overview of biosensing applications using DTs. First, the design and working principle of DTs are introduced. Next, the signal producing principles of DTs are summarized. Furthermore, biosensing applications of DTs for varying targets and purposes, both in buffers and complex biological environments, are highlighted. Finally, we provide potential opportunities and challenges for the further development of DTs.

Intravesical instillation has been widely utilized for bladder cancer treatment in clinic. However, due to the bladder mucosal barrier, its poor penetration efficiency and drug utilization limit the clinical therapeutic effectiveness and result in a high recurrence rate. Therefore, designing an efficient and controllable drug delivery nanoplatform is of great significance for bladder cancer treatment. Non-invasive therapy based on near-infrared-II (NIR-II) photothermal therapy (PTT) conduces to overcome bladder mucosal barrier and enhance drug delivery. Also, the photothermal nanomaterials, Au Hollow Nanorods (AuHNRs), demonstrate strong photothermal properties and drug loading capacity. Herein, a quaternized chitosan

While checkpoint blockade immunotherapy as a promising clinical modality has revolutionized cancer treatment, it is of benefit to only a subset of patients because of the tumor immunosuppressive microenvironment. Herein, we report that the specified delivery of vitamin C at the tumor site by responsive lipid nanoparticles can efficiently induce oxidative toxicity and the polarization of M1 macrophages, promoting the infiltration of activating cytotoxic T lymphocytes in the tumor microenvironment for intensive immune checkpoint blocking therapy. Both

The second near-infrared (NIR-II)-induced photothermal therapy (PTT) has attracted a great deal of attention in recent years due to its non-invasiveness and because it uses less energy. However, the penetration of photothermal agents into solid tumors is seriously impeded by the dense-tumor extracellular matrix (ECM) containing cross-linked hyaluronic acid (HA), thereby compromising the ultimate therapeutic effects. Herein, acid-labile metal-organic frameworks were employed as nanocarriers to efficiently mineralize hyaluronidase (HAase) and encapsulate Ag

Delivery of small interfering RNA (siRNA) to intact plants for gene silencing mainly relies on viral vectors and Agrobacterium-mediated transformation due to the barrier of intact plant cell wall. Here, we reported that polymer functionalized graphene oxide nanoparticles (GONs) enable siRNA transfer into intact plant cells and bring about efficient gene silencing. We found that sheeted GONs could efficiently load siRNA to form small sized, near-spheroidal GONs-siRNA complex, which could be across the cell wall and internalize in the plant cell. The GONs-siRNA exhibited transient and strong silencing (97.2 % efficiency) in plant tissues at 24 h after treatment and returned to normal level at 5 days after treatment. This method has the obvious advantages of efficient, transient, simple, stability and well biocompatibility, which should greatly stimulate the application of nanomaterials as gene-engineering tools in plant research.

Metal ions play critical roles in chemical, biological, and environmental processes. Various biomolecules have the ability to coordinate with metal ions and form various materials. Nucleobases, nucleosides, and nucleotides, as the essential components of DNA, have emerged as a useful building block for the construction of functional nanomaterials. In recent years, DNA oligonucleotides have also been used for this purpose. We herein review the strategies for the synthesis of soft nanomaterials through the assembly of nucleotides (or DNA) and metal ions to yield various nanoparticles, fibers, and hydrogels. Such coordination methods are simple to operate and can be carried out under ambient conditions. The luminescent, catalytic, and molecular recognition properties of these coordination materials are described with representative recent examples. Their applications ranging from biosensing, enzyme encapsulation, catalysis, templated shell growth to cancer therapy are highlighted. Finally, challenges of this field and future perspectives are discussed.

The sensitive detection of coronavirus is of vital importance for the prevention of its rapid spread. Porcine epidemic diarrhea virus (PEDV) is a highly contagious coronavirus that causes severe diarrhea and death in neonatal piglets. In this work, a novel PCN-224-based electrochemiluminescence (ECL) system was constructed for PEDV detection with high sensitivity. We found that PCN-224 can be employed as an ECL reporter with a strong signal because of its zirconium-based organic porous frame nanomaterial with a large specific surface area and stable structure. TiO2 nanoparticles were used as an accelerator for the first time to promote the reduction of coreactant potassium peroxydisulfate on the cathode; thus, the initial ECL signal of PCN-224 was significantly amplified. In the presence of PEDV, the ECL signal decreased due to the block effect to electron transfer. As a result, the novel "signal off" biosensor achieved a sensitive detection of PEDV ranging from 1 pg/mL to 10 ng/mL, with a detection limit of 0.4 pg/mL (S/N = 3). Importantly, the PCN-224 nanomaterial enriched the ECL system in biological analysis, and the proposed strategy provided a new route for coronavirus detection.

African swine fever (ASF) is an infectious disease caused by African swine fever virus (ASFV) with clinical symptoms of high fever, hemorrhages and high mortality rate, posing a threat to the global swine industry and food security. Quarantine and control of ASFV is crucial for preventing swine industry from ASFV infection. In this study, a recombinase polymerase amplification (RPA)-CRISPR-based nucleic acid detection method was developed for diagnosing ASF. As a highly sensitive method, RPA-CRISPR can detect even a single copy of ASFV plasmid and genomic DNA by determining fluorescence signal induced by collateral cleavage of CRISPR-lwCas13a (previously known as C2c2) through quantitative real-time PCR (qPCR) and has the same or even higher sensitivity than the traditional qPCR method. A lateral flow strip was developed and used in combination with RPA-CRISPR for ASFV detection with the same level of sensitivity of TaqMan qPCR. Likewise, RPA-CRISPR is capable of distinguishing ASFV genomic DNA from viral DNA/RNA of other porcine viruses without any cross-reactivity. This diagnostic method is also available for diagnosing ASFV clinical DNA samples with coincidence rate of 100% for both ASFV positive and negative samples. RPA-CRISPR has great potential for clinical quarantine of ASFV in swine industry and food security. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12250-020-00323-1) contains supplementary material, which is available to authorized users.

Biological nitrogen fixation is essential to crop production, but it can be inhibited by high concentrations of reactive oxygen species (ROS) due to the irreversible inactivation of the nitrogenase complex, thus posing a threat to food security and sustainable agricultural development. Here, we constructed an antioxidant cobalt ferrite (CoFe2O4) nanozyme as a bridge between nanotechnology and biological nitrogen fixation, which was shown to efficiently regulate the reactive oxygen metabolism and protect nitrogenase, thereby significantly enhancing the symbiotic nitrogen fixation efficiency by 260% in Glycine max (L.) Merr. (soybean). The CoFe2O4 nanozyme was also revealed to effectively reduce the concentration of ROS in the nodule by 56.6%, creating a superior environment for the proliferation of rhizobia and forming more effective nodules (larger nodules for an increase of 45.6% in the number of parasitic rhizobia). Furthermore, the CoFe2O4 nanozyme was shown to act as a synergist of leghemoglobin and increase its accumulation by 45.9%, where the high concentration of leghemoglobin in nodular cells can create a relatively hypoxic environment and protect nitrogenase, thus achieving a quantitative leap in nitrogen fixation capacity and simultaneously increasing the soybean photosynthesis by 67.2%. Our study has demonstrated that the CoFe2O4 nanozyme can efficiently regulate the intracellular ROS metabolism and serve as a promising strategy for enhancing symbiotic nitrogen fixation.

An ultrasensitive electrochemiluminescence (ECL) biosensor was developed based on MIL-53(Al)@CdS QDs and SiO2@AuNPs for trichlorfon detection. Metal–organic frameworks (MOFs) were used as a loading platform that provided a large surface area to load targets and modified materials onto the electrode. At the same time, SiO2@AuNPs loaded plenty of AuNPs which effectively increased the ECL resonance energy transfer between the CdS QDs, so that the ECL signal was strongly quenched and resulted in an amplified response. In the range of 10−11–10−4 M, the ECL response showed a linear relationship with the concentration (logarithm) of trichlorfon, and the detection limit was 5.1 × 10−12 M (S/N = 3). When the biosensor was applied to detect trichlorfon in lettuce, broccoli, cucumber, and chives, the recoveries obtained from the spiked samples were 97%–105%, 102%–104%, 100%–104%, and 98%–104%, respectively. Thus, this novel ECL biosensor has potential applications for the analysis of trichlorfon in food samples.

Nucleic acid lateral flow sensing has drawn great research attention since it has the advantages of being simple, rapid, and cost-effective. However, considering the trace amounts of the nucleic acid targets, its sensitivity is still limited. Although enormous efforts have been devoted to enhancing its sensitivity, developing a simple lateral flow sensing platform with high sensitivity remains challenging. We report a novel lateral flow microRNA-21 biosensing platform based on a portable surface enhanced Raman scattering (SERS) reader coupled with a catalytic hairpin assembly signal amplification strategy. Hairpin DNA probes were anchored on Au@Ag nanotags, and the presence of microRNA-21 triggered the formation of numerous double-stranded DNAs along with the exposure of the biotin groups. By this means, the target was recycled and signal amplification was achieved. The Au@Ag nanoprobes with exposed biotin can be captured on the test line via its interaction with streptavidin. By scanning the strip with a portable SERS reader, the sensitive quantification of microRNA-21 was realized with a detection limit as low as 84 fM. The proposed strategy was employed to detect the target in a serum sample, demonstrating its great potential in amplified point-of-care biosensing for clinical diagnosis.

With the wide application of nanomaterials (NMs) in agriculture, it is particularly important to assess the impact of these NMs on soil microorganisms. In this study, different varieties of soybean rhizosphere microorganisms (RM) were employed to simulate the alleviate effect of molybdenum nanoparticles (Mo NPs) induced stress in presence of soybean plants. Mo NPs caused serious toxic effects on soybean growth and nitrogen fixation at a concentration of 100 mg kg

The existence of contaminants in food poses a serious threat to human health. In recent years, aptamer sensors (aptasensors) have been developed rapidly for the detection of food contaminants because of their high specificity, design flexibility, and high efficiency. However, the development of high-throughput, highly sensitive, on-site, and cost-effective methods for simultaneous detection of food contaminants is still restricted due to multiple signal overlap or mutual interference and cross-reaction between different analytes with similar molecular structures. To overcome these problems, this Review summarizes some effective strategies from the articles published in recent years about multiplexed aptasensors for the simultaneous detection of food contaminants. This work focuses on the application of multiplexed aptasensors to simultaneously detect antibiotics, pathogens, and mycotoxins in food. These aptasensors mainly contain fluorescent aptasensors, electrochemical aptasensors, surface-enhanced Raman scattering-based aptasensors, microfluidic chip aptasensors, and paper-based multiplexed aptasensors. In addition, this Review also covers the application of nucleic acid cycle amplification and nanomaterial amplification strategies to improve the detection sensitivity. Finally, the limitations and challenges in the design of multiplexed aptasensor are also taken into account.

A short kanamycin-binding aptamer has been widely used for detecting kanamycin. One of the popular signaling methods is based on the color change of gold nanoparticles (AuNPs) to develop label-free colorimetric biosensors. The general perception was that aptamer binding to its target would inhibit aptamer adsorption by the AuNPs. This inhibited adsorption results in the aggregation of the AuNPs and a color change upon addition of salt. However, the potential adsorption of kanamycin was ignored. Herein, we carefully studied the adsorption of kanamycin on AuNPs and performed a comprehensive analysis using two mutated aptamers and a randomly sequenced DNA which were not supposed to bind kanamycin. In addition, a total of six antibiotics were studied over a wide concentration range. As low as 90 nM kanamycin can induce the aggregation of 3 nM citrate-capped AuNPs, indicating very strong adsorption of kanamycin. The color change was independent of DNA sequence, and all the tested sequences showed a similar color response, regardless of aptamer. Among the different antibiotics, kanamycin and streptomycin induced a color change but not the other four. Our results support an alternative mechanism that kanamycin and streptomycin adsorption by the AuNPs was the main reason for the color change instead of aptamer binding.

With the rapid development of nanotechnology, molybdenum (Mo)-based nanomaterials have been widely used in various fields, and the ensuing environmental behavior has also raised widespread concern....

Activated doxorubicin (DOX) often has severe systemic toxicity and side effects due to its inability to distinguish tumor cells from normal cells, which seriously affects the prognosis of patients. Here, we synthesized an inactivated a DOX prodrug that could be selectively activated by a light-induced caspase-3 enzyme in the tumor site. In the absence of light, this uniformly dispersed nanoparticle avoided the unnecessary toxicity under physiological conditions. Upon the laser irradiating to the tumor area of interest, the nanoparticles can produce a large amount of reactive oxygen species (ROS) to induce cell apoptosis and activate caspase-3 enzyme to release DOX selectively. Meanwhile, the produced ROS can also combine with activated DOX to cause more potent tumor damage. The experiments demonstrated that the light can effectively activate DOX drug through a series of cascade events and the subsequent synergistic therapy both

A DNA tweezer is a dynamic DNA nanomachine that can reversibly switch its state between open and closed. Here, we employed a DNA tweezer for the first time to dynamically control the distance between plasmonic silver nanoparticles (Ag NPs) for a surface enhanced Raman scattering (SERS) biosensing application. Two DNA and 4-nitrothiophenol (4-NTP) modified Ag NPs were linked to the arms of the DNA tweezer (DNA tweezer-Ag NPs probe) by complementary base pairing. Activation of the Raman intensity was achieved by the state transformation of the DNA tweezer-Ag NPs probe from open to closed. The distances between two Ag NPs in open and closed state were 8.1 ± 2.7 nm and 3.2 ± 0.8 nm, respectively. Furthermore, the two Ag NPs were spatially separated in the open state with a low Raman signal, whereas in the closed state, Raman intensity was enhanced because of the proximity of two Ag NPs. The developed biosensing system exhibited a good linear relationship when the concentration of aflatoxin B1 (AFB1) ranged from 1 ng/mL to 0.01 pg/mL, and the limit of detection (LOD) was 5.07 fg/mL. In addition, spike recovery and certificated real foodstuffs were used to examine the feasibility in a real situation. This protocol provides a potential candidate for SERS detection and can be used as a promising technology for biological and chemical sensors.

An aptasensor based on MIL-53(Al)@CdTe was designed for multiple determination of Hg2+ and Pb2+ by electrochemiluminescence (ECL). Upon the recognition of Hg2+, aptamer 2-AuNPs form hairpin structures and are removed from the electrode. While in the presence of Pb2+, aptamer 1-PtNPs capture the target ions and form G-quadruplexes, and then bring PtNPs close enough to CdTe QDs to produce ECL resonance energy transfer. Upon aptamer interaction with Hg2+ and Pb2+, decreased ECL intensity was observed due to enhanced resonance energy transfer (ERET) and attenuated surface plasmon resonance (SPR). The ECL intensity difference (ΔECL) could therefore be used to detect heavy-metal ions with detection limits of 4.1 × 10−12 M (path 1, Hg2+), 3.7 × 10−11 M (path 2, Pb2+), and 2.4 × 10−11 M (path 3, Pb2+). The aptasensor could also be used for detecting Hg2+ and Pb2+ in fish and shrimp samples with good recoveries.

The CRISPR/Cas9 mediated genome editing have provided a promising strategy to correct multiple mutations of Duchenne muscular dystrophy (DMD). However, the delivery of CRISPR/Cas9 system into mammalian cell for DMD gene editing mainly relies on adeno associated virus (AAV)-mediated transport. Meanwhile, the protospacer adjacent motif (PAM) requirement of wild-typed Cas9 protein causing the target sites for exon splice acceptor site are restricted to limited regions. Here, we developed a biomineralized PAMLess Cas9 (SpRY) variant nanoparticles (Bm-SpRY NPs) for DMD gene editing in vitro and in vivo. This method described a facile synthesis of biomineralized NPs with high SpRY pDNA encapsulation efficiency. In vitro results show that the Bm-SpRY NPs have the obvious advantages of well biocompatibility and protecting SpRY pDNA from enzyme degradation and efficient delivery under high serum condition. Cell studies demonstrated that Bm-SpRY NPs enable rapid cellular uptake, endo-lysosomes escape and nucleus transport. Meanwhiles, the DMD gene editing via Bm-SpRY NPs pathway is transient process without genomic integration. We evaluated multiple target regions with different PAMs for the DMD exon 51 splice acceptor site through Bm-SpRY NPs method and found that the target region with TAG PAM has the highest editing efficiency and significant preferential mutation. In vivo results show that intramuscular injection of Bm-SpRY NPs enable DMD gene mutation in muscle tissue without tissue damage. This study may extend the advanced application of CRISPR system for DMD therapy. STATEMENT OF SIGNIFICANCE: The gene editing technology of CRISPR/Cas9 provides an effective treatment strategy for the Duchenne muscular dystrophy (DMD) therapy. However, the delivery of CRISPR system in mammalian cell mainly relies on viral mediated transport and the NGG or NAG requirement of wild-typed Cas9 protein limits the target region in DMD gene. Here, the present study provides a biomineralized PAM Less Cas9 (SpRY) variant nanoparticles (Bm-SpRY NPs) for DMD gene editing in vitro and in vivo. This study may extend the application of CRISPR system for DMD gene therapy.

Infection with

Metal-respiring bacteria are frequently used to recycle metal resources by biosynthesizing nanoparticles on its surface in environment treatment. However, further utilization of biogenetic nanoparticles through combining the advantages of both bacteria and nanoparticles is still limited. Herein, biogenetic Au@Ag nanoislands are utilized as the surface-enhanced Raman spectroscopy (SERS) substrate for quantitative detection. Specifically, Au@Ag nanoislands enhance the Raman signal

Developing nanomaterials-based antimicrobial agents has shown a widespread promise. In this study, silver nanoparticle-modified graphene oxide (GO-AgNPs) nanocomposites were self-assembled via interfacial electrostatic force. By using the porcine reproductive and respiratory syndrome virus (PRRSV) as a pattern, the antiviral effect of the as-prepared GO-AgNPs nanocomposites on the replication of virus was investigated. The results indicated that exposure with GO-AgNPs nanocomposites could obviously suppress PRRSV infection. It was found that GO-AgNPs nanocomposites exhibited a better inhibitory effect compared with AgNPs and GO. By selecting the porcine epidemic diarrhea virus (PEDV) as a contrast virus, GO-AgNPs nanocomposites were proven to have a broad antiviral activity. Mechanism studies showed that GO-AgNPs nanocomposites might prevent PRRSV from entering the host cells, with 59.2% inhibition efficiency. Meanwhile, GO-AgNPs nanocomposite treatment enhances the production of interferon-α (IFN-α) and IFN-stimulating genes (ISGs), which can directly inhibit the proliferation of virus. Taken together, this study reports a new type of antiviral agent and provides a promising pharmaceutical agent for treating infection by the highly pathogenic PRRSV. Moreover, it may provide novel ideas for the research and development of antiviral formulations based on nanocomposites and extend their applications in biological systems.

Mitochondria play critical roles in the regulation of the proliferation and apoptosis of cancerous cells. Targeted induction of mitochondrial dysfunction in cancer cells by multifunctional nanosystems for cancer treatment has attracted increasing attention in the past few years. Numerous therapeutic nanosystems have been designed for precise tumor therapy by inducing mitochondrial dysfunction, including reducing adenosine triphosphate, breaking redox homeostasis, inhibiting glycolysis, regulating proteins, membrane potential depolarization, mtDNA damage, mitophagy dysregulation and so on. Understanding the mechanisms of mitochondrial dysfunction would be helpful for efficient treatment of diseases and accelerating the translation of these therapeutic strategies into the clinic. Then, various strategies to construct mitochondria-targeted nanosystems and induce mitochondrial dysfunction are summarized, and the recent research progress regarding precise tumor therapeutics is highlighted. Finally, the major challenges and an outlook in this rapidly developing field are discussed. This review is expected to inspire further development of novel mitochondrial dysfunction-based strategies for precise treatments of cancer and other human diseases.

Rabies is caused by the infection of Rabies virus, it leads to fatal encephalitis, developing a highly sensitive and specific detection method for Rabies virus remains a challenge. Herein, we report an electrochemiluminescence (ECL) biosensor for Rabies viral RNA based on dual-signal amplification and DNA nanotweezers (DTs). Dual-signal amplification process includes target binding induced isothermal amplification and CRISPR-based amplification. In the presence of target RNA, two assisted probes simultaneously hybridized with it to trigger isothermal amplification with the help of polymerase and nicking enzyme. This process generated a large amount of single-stranded DNA (ssDNA) as products. The products hybridized with CRISPR RNA to activate the trans-cleavage activity of Cas12a to indiscriminately cleave predesigned single-stranded trigger (ST) strands. After mixing the cleavage products with DTs and hemin molecules, DTs cannot be closed by cleaved ST strands to capture hemin to the electrode to quench the ECL signal. Therefore, the higher concentration of the target, the stronger intensity of the ECL signal. The detection limit is as low as 2.8 pM and the detection range is from 5 pM to 5 nM with excellent specificity and stability. The proposed method provides a promising strategy for Rabies detection, and can be easily adapted to other analytes via reasonable design as a valuable and versatile tool in bioanalysis.

Due to the high recurrence rate and mortality of venous thrombosis, there is an urgent need for research on antithrombotic strategies. Because of the short half-life, poor targeting capabilities, bleeding complications, and neurotoxic effects of conventional pharmacological thrombolysis methods, it is essential to develop an alternative strategy to noninvasive thrombolysis and decrease the recurrence rate of venous thrombosis. A platelet-mimetic porphyrin-based covalent organic framework-engineered melanin nanoplatform, to target delivery of hirudin to the vein thrombus site for noninvasive thrombolysis and effective anticoagulation, is first proposed. Owing to the thrombus-hosting properties of platelet membranes, the nanoplatform can target the thrombus site and then activate hyperthermia and reactive oxygen species for thrombolysis under near-infrared light irradiation. The photothermal therapy/photodynamic therapy combo can substantially improve the effectiveness (85.7%) of thrombolysis and prevent secondary embolism of larger fragments. Afterward, the highly loaded (97%) and slow-release hirudin (14 days) are effective in preventing the recurrence of blood clots without the danger of thrombocytopenia. The described biomimetic nanostructures offer a promising option for improving the efficacy of thrombolytic therapy and reducing the risk of bleeding complications in thrombus associated diseases.

One of the main challenges for tumor vascular infarction in combating cancer lies in failing to produce sustained complete thrombosis. Inspired by the capability of vascular infarction in blocking the delivery of oxygen to aggravate tumor hypoxia, we herein present the performance of selective tumor thrombus inducing hypoxia activation therapy to improve the therapeutic index of coagulation-based tumor therapy. By encapsulating coagulation-inducing protease thrombin and a hypoxia-activated prodrug (HAP) tirapazamine into metal-organic framework nanoparticles with a tumor-homing ligand, the obtained nanoplatform selectively activates the platelet aggregation at tumor to induce the thrombosis and vascular obstruction therapy by the exposed thrombin. Meanwhile, the thrombus can cut off the blood oxygen supply and potentiate the hypoxia levels to enhance the HAP therapy. This strategy not only addresses the dissatisfaction of vascular therapy, but also conquers the dilemma of inadequate hypoxia in the HAP treatment. Since clinical operations such as surgery can be used to induce the coagulation, the coagulation based synergistic therapy is promising to be translated into a clinical combination regimen. Agency for Science, Technology and Research (A*STAR) National Research Foundation (NRF) Submitted/Accepted version This work was supported by grants from the National Natural Science Foundation of China (21778020, 31750110464, and 31950410755), the Sci-Tech Innovation Foundation of Huazhong Agricultural University (2662018PY024), the National Key R&D Program of China (2016YFD0500706), and the Science and Technology Major Project of Guangxi (Gui Ke AA18118046). This work was also supported by the Singapore Agency for Science, Technology and Research (A*STAR) AME IRG grant (A20E5c0081), and the Singapore National Research Foundation Investigatorship (NRF-NRFI2018-03).

Delivery of the CRISPR/Cas9 ribonucleoprotein (RNP) complex has provided an alternative strategy for the regulation of CRISPR functions, offering a transient and DNA-free means for genomic editing. Chemical methods of delivering the RNPs via nanocomplexes have the potential to address these delivery problems for efficiency, safety, and packaging capacity. Here, we developed a biomimetic mineralization-mediated strategy for efficient DNA-free genome editing by using CRISPR/Cas9 RNPs. We found that the RNPs have the ability to form the biomimetic mineralized RNP nanoparticles (Bm-RNP NPs) quickly in situ and can be effectively delivered into the fungal protoplast cells. Biomimetic mineralization can maintain the natural function of Cas9 protein and protect the sgRNA activity. At the same time, the encapsulated RNPs can be effectively released into the cytoplasm, and the Sytalone dehydratase (SDH) gene can be edited in a targeted manner. Except for phenotypic defects, molecular detections indicated that the delivery of Bm-RNP NPs achieved 20% genomic editing for Magnaporthe oryzae compared to RNPs alone. Moreover, the Bm-RNP NP-mediated editing of the SDH gene significantly affects the appressorium-mediated penetration and invasive growth in M. oryzae. Our system has the advantages of being cheap, fast, and effective, without the traditional transformation process, suggesting the potential application of this DNA-free gene-editing strategy in different organisms.