Programmably engineered stochastic RNA nanowalker for ultrasensitive miRNA detection.

A programmably engineered stochastic RNA nanowalker powered by duplex-specific nuclease (DSN) is developed. By utilizing poly-adenine-based spherical nucleic acids (polyA-SNA) to accurately regulate the densities of DNA tracks, the nanowalker showcases its capability to identify miRNA-21, miRNA-486, and miRNA-155 with quick kinetics and attomolar sensitivity, positioning it as a promising option for cancer clinical surveillance.

Enhancing Point-of-Care Diagnosis of African Swine Fever Virus (ASFV) DNA with the CRISPR-Cas12a-Assisted Triplex Amplified Assay.

Accurate, ultrasensitive, and point-of-care (POC) diagnosis of the African swine fever virus (ASFV) remains imperative to prevent its spread and limit the losses incurred. Herein, we propose a CRISPR-Cas12a-assisted triplex amplified colorimetric assay for ASFV DNA detection with ultrahigh sensitivity and specificity. The specific recognition of recombinase aided amplification (RAA)-amplified ASFV DNA could activate the Cas12a/crRNA/ASFV DNA complex, leading to the digestion of the linker DNA (bio-L1) on magnetic beads (MBs), thereby preventing its binding of gold nanoparticles (AuNPs) network. After magnetic separation, the release of AuNPs network comprising a substantial quantity of AuNPs could lead to a discernible alteration in color and significantly amplify the plasmonic signal, which could be read by spectrophotometers or smartphones. By combining the RAA, CRISPR/Cas12a-assisted cleavage, and AuNPs network-mediated colorimetric amplification together, the assay could detect as low as 0.1 copies/µL ASFV DNA within 1 h. The assay showed an accuracy of 100% for the detection of ASFV DNA in 16 swine tissue fluid samples, demonstrating its potential for on-site diagnosis of ASFV.

A three-in-one point-of-care electrochemical sensing platform for accurate monitoring of diabetes.

A three-in-one electrochemical sensing platform was designed for the simultaneous detection of total hemoglobin (tHb), glycated hemoglobin (HbA1c) and HbA1c% by using a dual-aptamer sensing strategy. The developed sensing platform exhibits excellent sensitivity, selectivity, repeatability and long-term stability, and holds promising prospects in the early diagnosis and long-term monitoring of diabetes.

Syntheses, crystal structures and MMCT properties of diruthenium-based cyanido-bridged RuV/VI2-NC-RuII complexes.

The goal of this study was to investigate how the electron-donating capability around the lower valent metal ion and the electron-accepting capability of the higher valent metal ion influence metal to metal charge transfer (MMCT) properties in mixed-valence complexes. A series of trinuclear ruthenium complexes represented as [Ru2(ap-4-Me)3(CH3COO)NCRuCpMex(dppe)][PF6] (CpMex = polymethylcyclopentadienyl, x = 0, 1, and 5; and dppe = 1, 2-bis(diphenylphosphino)ethane, ap-4-Me = 2-anilino-4-methylpyridine) and their one-electron oxidized products were synthesized and fully characterized. The UV-vis-NIR spectra confirmed that as the electron donor character of the CpMex(dppe)RuCN fragment enhanced or the electron-accepting capability of the higher valent diruthenium cluster increased, the RuII → RuV2 or RuVI2 Ru2 MMCT bands shifted to lower energies, which was supported by TDDFT calculations.

Short-term and long-term effectiveness of acupuncture and Tuina on knee osteoarthritis: study protocol for a randomized controlled trial.

Background: The effectiveness of acupuncture and tuina in treating knee osteoarthritis (KOA) is still controversial, which limits their clinical application in practice. This study aims to evaluate the short-term and long-term effectiveness of acupuncture and tuina on KOA. Methods/design: This parallel-group, multicenter randomized clinical trial (RCT) will be conducted at the outpatient clinic of five traditional Chinese medicine hospitals in China. Three hundred and thirty participants with KOA will be randomly assigned to acupuncture, tuina, or home-based exercise group with a ratio of 1:1:1. The primary outcome is the proportion of participants achieving a minimal clinically important improvement defined as a ≥ 12% reduction on the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain dimension on short term (week 8) and long term (week 26) compared with baseline. Secondary outcomes are knee joint conditions (pain, function, and stiffness), self-efficacy of arthritis, quality of life, and psychological conditions, which will be evaluated by the WOMAC score and the Patient Global Assessment (PGA), and in addition, the respondents index of OMERACT-OARSI, Short Form 12 Health Survey (SF-12), arthritis self-efficacy scale, and European five-dimensional health scale (EQ-5D). Adverse events will be collected by self-reported questionnaires predefined. Clinical trial registration: https://www.chictr.org.cn.

Construction of a point-of-care electrochemical biosensor for Escherichia coli 16S rRNA analysis based on MoS2 nanoprobes.

Foodborne pathogens are harmful to human health because they can contaminate food and induce diseases. To efficiently distinguish and determine foodborne bacteria, an ultrasensitive point-of-care electrochemical biosensor was designed for 16S rRNA detection by coupling a signal amplification strategy with MoS2-based nanoprobes. Gold nanoparticles and thionine co-functionalized molybdenum disulfide (MoS2) nanocomposites (MoS2-Thi-AuNPs) were used to construct nanoprobes, which can efficiently monitor the detection process and amplify the detection signal. In the presence of Escherichia coli (E. coli) 16S rRNA, a classical sandwiched DNA structure was formed on the surface of a hierarchical flower-like gold nanostructure-decorated screen-printed carbon electrode (HFGN-SPCE), generating an obvious electrochemical signal from Thi. Under optimal conditions, this designed electrochemical biosensor showed a wide dynamic range (0-1.0 × 106 fM), low detection limit (2.8 fM), high selectivity and accepted stability for E. coli 16S rRNA detection in ideal buffers. Moreover, this biosensor can efficiently determine 16S rRNA in milk samples and 10 CFU mL-1 bacterial lysate. All experimental results suggested that this biosensor has a promising application in the detection of foodborne pathogens.

Reverse genetic study reveals the molecular targets of chordotonal organ TRPV channel modulators.

Insecticides have been widely used for the control of insect pests that have a significant impact on agriculture and human health. A better understanding of insecticide targets is needed for effective insecticide design and resistance management. Pymetrozine, afidopyropen and flonicamid are reported to target on proteins that located on insect chordotonal organs, resulting in the disruption of insect coordination and the inhibition of feeding. In this study, we systematically examined the susceptibility of six Drosophila melanogaster mutants (five transient receptor potential channels and one mechanoreceptor) to three commercially used insecticides, in order to identify the receptor subunits critical to the insect's response to insecticides. Our results showed that iav1, nan36aand wtrw1 mutants exhibited significantly reduced susceptibility to pymetrozine and afidopyropen, but not to flonicamid. The number of eggs produced by the three mutant females were significantly less than that of the w1118 strain. Meanwhile, the longevity of all male mutants and females of nan36a and wtrw1 mutants was significantly shorter than that of the w1118 strain as the control. However, we observed no gravitaxis defects in wtrw1 mutants and the anti-gravitaxis of wtrw1 mutants was abolished by pymetrozine. Behavioral assays using thermogenetic tools further confirmed the bioassay results and supported the idea that Nan as a TRPV subfamily member located in Drosophila chordotonal neurons, acting as a target of pymetrozine, which interferes with Drosophila and causes motor deficits with gravitaxis defects. Taken together, this study elucidates the interactions of pymetrozine and afidopyropen with TRPV channels, Nan and Iav, and TRPA channel, Wtrw. Our research provides another evidence that pymetrozine and afidopyropen might target on nan, iav and wtrw channels and provides insights into the development of sustainable pest management strategies.

Combined VEGF and bFGF loaded nanofiber membrane protects against neuronal injury and hypomyelination in a rat model of chronic cerebral hypoperfusion.

Currently, there are no effective therapeutic targets for the treatment of chronic cerebral hypoperfusion(CCH)-induced cerebral ischemic injury. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are discovered as the inducers of neurogenesis and angiogenesis. We previously made a nanofiber membrane (NFM), maintaining a long-term release of VEGF and bFGF up to 35 days, which might make VEGF and bFGF NFM as the potential protective agents against cerebral ischemic insult. In this study, the effects of VEGF and bFGF delivered by NFM into brain were investigated as well as their underlying mechanismsin a rat model of CCH. VEGF + bFGF NFM application increased the expressions of tight junction proteins, maintained BBB integrity, and alleviated vasogenic cerebral edema. Furthermore, VEGF + bFGF NFM sticking enhanced angiogenesis and elevated CBF. Besides, VEGF + bFGF NFM treatment inhibited neuronal apoptosis and decreased neuronal loss. Moreover, roofing of VEGF + bFGF NFM attenuated microglial activation and blocked the launch of NLRP3/caspase-1/IL-1ß pathway. In addition, VEGF + bFGF NFM administration prevented disruption to the pre/postsynaptic membranes and loss of myelin sheath, relieving synaptic injury and demyelination. Oligodendrogenesis, neurogenesis and PI3K/AKT/mTOR pathway were involved in the treatment of VEGF + bFGF NFM against CCH-induced neuronal injury and hypomyelination. These findings supported that VEGF + bFGF NFM application constitutes a neuroprotective strategy for the treatment of CCH, which may be worth further clinical translational research as a novel neuroprotective approach, benifiting indirect surgical revascularization.

Enhanced electrochemiluminescence imaging of single cell membrane proteins based on Co3O4 nanozyme catalysis.

The development of enhanced strategies with excellent biocompatibility is critical for electrochemiluminescence (ECL) imaging of single cells. Here, we report an ECL imaging technique for a single cell membrane protein based on a Co3O4 nanozyme catalytic enhancement strategy. Due to the remarkable catalytic performance of Co3O4 nanozymes, H2O2 can be efficiently decomposed into reactive oxygen radicals, and the reaction with L012 was enhanced, resulting in stronger ECL emission. The anti-carcinoembryonic antigen (CEA) was coupled with nanozyme particles to construct a probe that specifically recognized the overexpressed CEA on the MCF-7 cell membrane. According to the locally enhanced visualized luminescence, the rapid ECL imaging of a single cell membrane protein was eventually realized. Accordingly, Co3O4 nanozymes with highly efficient activity will provide new insights into ECL imaging analysis of more biological small molecules and proteins.

Synthesis of Graphene Oxide-Coupled CoNi Bimetallic MOF Nanocomposites for the Simultaneous Analysis of Catechol and Hydroquinone.

Herein, a three-dimensional flower-like cobalt-nickel bimetallic metal-organic framework (CoNi-MOF) coupled with two-dimensional graphene oxide (GO) nanocomposites was successfully synthesized for the selective and simultaneous electrochemical determination of catechol (CC) and hydroquinone (HQ). The three-dimensional flower-like structure of the CoNi-MOF/GO nanocomposite has a multilayer structure and a large surface area, which greatly improves its electrocatalytic activity towards CC and HQ. Differential pulse voltammetry (DPV) results showed that the peak-to-peak separation of CC (0.223 V) and HQ (0.120 V) was 103 mV at a CoNi-MOF/GO modified glassy carbon electrode (CoNi-MOF/GO/GCE), suggesting that the proposed modified electrode can selectively and simultaneously determine them. Under optimal conditions, the CoNi-MOF/GO/GCE showed an excellent analytical performance for the simultaneous determination of CC and HQ, including a wide linear range (0.1-100 µM), low detection limit (0.04 µM for HQ and 0.03 µM for CC) and high anti-interference ability. As expected, the developed modified electrode has been used to analyze CC and HQ in river water, with acceptable results.

Biological Recognition-Based Electrochemical Aptasensor for Point-of-Care Detection of cTnI.

As a "gold standard biomarker", cardiac troponin I (cTnI) is widely used to diagnose acute myocardial infarction (AMI). For an early clinical diagnosis of AMI, it is necessary to develop a facile, fast and on-site device for cTnI detection. According to this demand, a point-of-care electrochemical aptasensor was developed for cTnI detection by coupling the advantages of screen-printed carbon electrode (SPCE) with those of an aptamer. Thiol and methylene blue (MB) co-labelled aptamer (MB-Apt-SH) was assembled on the surface of hierarchical flower-like gold nanostructure (HFGNs)-decorated SPCE (SPCE-HFGNs) to recognize and analyze cTnI. In the presence of cTnI, the specific biological recognition reaction between cTnI and aptamer caused the decrease in electrochemical signal. Under the optimal condition, this designed aptasensor showed wide linear range (10 pg/mL-100 ng/mL) and low detection limit for (8.46 pg/mL) for cTnI detection with high selectivity and stability. More importantly, we used a mobile phone coupled with a simple APP to efficiently detect cTnI in 10 µL 100% human serum samples, proving that this aptasensor has a promising potential in point-of-care testing.

Electronic Transition and Magnetic Coupling Regulation in Trimetallic Complexes Featuring a New Bridging Ligand Obtained by Oxidative Addition.

A series of trimetallic complexes [FeIII(µ-L)(py)]2MII(py)n (n = 2, MII = MnII, 1; FeII, 2; CoII, 3; ZnII, 4; n = 3, MII = CdII, 5) with a new bridging ligand L4- (deprotonated 1,2-N1,N2-bis(2-mercaptoanil) oxalimidic acid) were synthesized and fully characterized by elemental analysis, single-crystal X-ray crystallography, IR, and Mössbauer spectra. Interestingly, the bridging ligand was obtained by oxidative addition of the (gma•)3- ligand from the mononuclear precursor Fe(gma)py (gma = glyoxal-bis(2-mercaptoanil)). In the obtained complexes, the bridging ligand L4- coordinates to the terminal FeIII ions (intermediate-spin with SFe = 3/2) by the N, S atoms, and coordinate to the central metal MII ion by the four O atoms. The resonance structure of the bridging ligand can be described as the two 4π-electron delocalized systems connected by one single-bond (C1-C2), which is different from the electronic structure of the precursor Fe(gma)py. Remarkably, the magnetic coupling interaction can be regulated through the central metal. The ferromagnetic coupling constant J gradually decreases as MII changes from FeII to CoII and MnII, while the paramagnetic behaviors are presented when MII = ZnII and CdII, confirmed by the magnetic susceptibility measurements and further supported by using the PHI program. Furthermore, the bridging ligand to the terminal FeIII charge transfer (LMCT) transitions emerged in all complexes but the central FeII to terminal FeIII charge transfer (MMCT) only presented in complex 2, strongly supported by the UV/vis-NIR electronic spectra and TDDFT calculations.

Layer-by-Layer Microneedle-Mediated rhEGF Transdermal Delivery for Enhanced Wound Epidermal Regeneration and Angiogenesis.

Appropriate treatments for acute traumas tend to avoid hemorrhages, vascular damage, and infections. However, in the homeostasis-imbalanced wound microenvironment, currently developed therapies could not precisely and controllably deliver biomacromolecular drugs, which are confronted with challenges due to large molecular weight, poor biomembrane permeability, low dosage, rapid degradation, and bioactivity loss. To conquer this, we construct a simple and effective layer-by-layer (LBL) self-assembly transdermal delivery patch, bearing microneedles (MN) coated with recombinant human epidermal growth factor (LBL MN-rhEGF) for a sustained release to wound bed driven by typical electrostatic force. Pyramidal LBL MN-rhEGF patches hold so enough mechanical strength to penetrate the stratum corneum, and generated microchannels allow rhEGF direct delivery in situ. The administrable delivery of biomacromolecular rhEGF through hierarchically coated MN arrays follows the diffusion mechanism of Fick's second law. Numerous efforts further have illustrated that finger-pressing LBL MN-rhEGF patches could not only promote cell proliferation of normal human dermal fibroblasts (NHDF) and human umbilical vein endothelial cells (HUVEC) in vitro but also take significant effects (regenerative epidermis: ∼144 µm; pro-angiogenesis: higher CD31 expression) in accelerating wound healing of mechanically injured rats, compared to the traditional dressing, which relies on passive diffusion. Our proof-of-concept features novel LBL biomacromolecular drug-delivery systems and self-administrated precision medicine modes at the point of care.

programmably engineered FRET-nanoflare for ratiometric live-cell ATP imaging with anti-interference capability.

Herein, we present a poly-adenine (polyA)-mediated programmably engineered FRET-nanoflare for ratiometric intracellular ATP imaging with anti-interference capability. The programmable polyA attachment is advantageous in enhancing the signal response for ATP. Moreover, the FRET-based nanoflare is capable of avoiding false-positive signals due to probe degradation in a complex environment, which has great potential for clinical diagnosis.

Spatiotemporal Monitoring of Subcellular mRNAs In Situ via Polyadenine-Mediated Dual-Color Sticky Flares.

Spatiotemporal monitoring of multiple low-abundance messenger RNAs (mRNAs) is vitally important for the diagnosis and pathologic analysis of cancer. However, it remains a clinical challenge to monitor and track multiple mRNAs location simultaneously in situ at subcellular level with high efficiency. Herein, we proposed polyA-mediated dual-color sticky flares for simultaneous imaging of two kinds of intracellular mRNA biomarkers. Two kinds of fluorescent DNA specific for GalNac-T mRNA and c-Myc mRNA were functionalized onto gold nanoparticles (AuNPs) through efficient polyadenine (polyA) attachment. By tuning polyA length, the lateral spacing and densities of DNA on AuNPs could be precisely engineered. Compared to the traditional thio-DNA-modified nanoprobes, the uniformity, detection sensitivity, and response kinetics of sticky flares were greatly improved, which enables live-cell imaging of mRNAs with enhanced efficiency. With a sticky-end design, the fluorescent DNA could dynamically trace mRNAs after binding with target mRNAs, which realized spatiotemporal monitoring of subcellular mRNAs in situ. Compared to one target mRNA imaging mode, the multiple target imaging mode allows more accurate diagnosis of cancer. Furthermore, the proposed polyA-mediated dual-color sticky flares exhibit excellent cell entry efficiency and low cytotoxicity with a low-cost and simple assembling process, which provide a pivotal tool for multiple targets imaging in living cells.

[Characterization of Ambient Volatile Organic Compounds, Source Apportionment, and the Ozone-NOx-VOC Sensitivities in Liucheng County, Guangxi].

Liucheng county, as a suburb of Liuzhou City in Guangxi province, has a prominent ozone (O3) pollution problem; however, there have been no relevant analyses of the cause of local O3 pollution reported. In order to investigate the causes of O3 pollution, an online observation of 116 VOCs with a time resolution of 1 h was carried out in Liucheng county from October 1st to 15th, and the sensitivity of ozone to the relative changes in the NOx and VOCs was analyzed. The results showed that the average value of φï¼»total volatile organic compounds (TVOCs)ï¼½ during the observation period was 27.52×10-9, and the average value of φ(TVOCs) during the polluting process (October 1st to 6th) was 32.15×10-9, which was 32.79% higher than that of the non-pollution process (October 8th to 15th). In terms of species concentration, oxygenated volatile organic compounds (OVOCs) contributed the highest, accounting for 43.70%, followed by alkanes (23.00%), aromatics (11.75%), and halocarbons (7.35%). In terms of ozone formation potential (OFP), OVOCs contributed the highest (41.96%) to OFP, followed by aromatics (32.60%) and alkenes (17.92%). During the observation period, VOCs mainly came from motor vehicle emissions (32.44%), biomass combustion sources (29.31%), solvent use sources (16.43%), plant sources (11.34%), and chemical industry emissions (10.49%). The contribution ratios of solvent use sources and plant sources in the pollution process increased by 28.58% and 28.53%, respectively. The EKMA curve shows that, during the observation period, Liucheng county was in a synergistic control area for VOCs and nitric oxide (NOx). Therefore, in the high ozone-occurrence autumn of Liucheng county, the key will be to reduce both VOCs and NOx emissions.

Effects of dietary oat supplementation on carcass traits, muscle metabolites, amino acid profiles, and its association with meat quality of Small-tail Han sheep.

Oat supplementation of the ruminant diet can improve growth performance and meat quality traits, but the role of muscle metabolites has not been evaluated. This study aimed to establish whether oat grass supplementation (OS) of Small-tail Han sheep improved growth performance and muscle tissue metabolites that are associated with better meat quality and flavor. After 90-day, OS fed sheep had higher live-weight and carcass-weight, and lower carcass fat. Muscle metabolomics analysis showed that OS fed sheep had higher levels of taurine, l-carnitine, inosine-5'-monophospgate, cholic acid, and taurocholic acid, which are primarily involved in taurine and hypotaurine metabolism, purine metabolism, and bile acid biosynthesis and secretion, decreased fat accumulation and they promote functional or flavor metabolites. OS also increased muscle levels of amino acids that are attributed to better quality and flavorsome mutton. These findings provided further evidence for supplementing sheep with oat grass to improve growth performance and meat quality.

Fecal microbiota transplantation and short-chain fatty acids protected against cognitive dysfunction in a rat model of chronic cerebral hypoperfusion.

AIMS: Clear roles and mechanisms in explaining gut microbial dysbiosis and microbial metabolites short-chain fatty acids (SCFAs) alterations in chronic cerebral ischemic pathogenesis have yet to be explored. In this study, we investigated chronic cerebral hypoperfusion (CCH)-induced gut microbiota and metabolic profiles of SCFAs as well as the effects and mechanisms of fecal microbiota transplantation (FMT) and SCFAs treatment on CCH-induced hippocampal neuronal injury. METHODS: Bilateral common carotid artery occlusion (BCCAo) was used to establish the CCH model. Gut microbiota and SCFAs profiles in feces and hippocampus were evaluated by 16S ribosomal RNA sequencing and gas chromatography-mass spectrometry. RNA sequencing analysis was performed in hippocampal tissues. The potential molecular pathways and differential genes were verified through western blot, immunoprecipitation, immunofluorescence, and ELISA. Cognitive function was assessed via the Morris water maze test. Ultrastructures of mitochondria and synapses were tested through a transmission electron microscope. RESULTS: Chronic cerebral hypoperfusion induced decreased fecal acetic and propionic acid and reduced hippocampal acetic acid, which were reversed after FMT and SCFAs administration by changing fecal microbial community structure and compositions. Furthermore, in the hippocampus, FMT and SCFAs replenishment exerted anti-neuroinflammatory effects through inhibiting microglial and astrocytic activation as well as switching microglial phenotype from M1 toward M2. Moreover, FMT and SCFAs treatment alleviated neuronal loss and microglia-mediated synaptic loss and maintained the normal process of synaptic vesicle fusion and release, resulting in the improvement of synaptic plasticity. In addition, FMT and SCFAs supplement prevented oxidative phosphorylation dysfunction via mitochondrial metabolic reprogramming. The above effects of FMT and SCFAs treatment led to the inhibition of CCH-induced cognitive impairment. CONCLUSION: Our findings highlight FMT and SCFAs replenishment would be the feasible gut microbiota-based strategy to mitigate chronic cerebral ischemia-induced neuronal injury.

VEGF loaded nanofiber membranes inhibit chronic cerebral hypoperfusion-induced cognitive dysfunction by promoting HIF-1a/VEGF mediated angiogenesis.

We investigated the potential effects and mechanisms of vascular endothelial growth factor (VEGF)-nanofiber membranes (NFMs) treatment in a rat model of chronic cerebral hypoperfusion (CCH). VEGF-NFMs treatment promoted angiogenesis in surgical temporal cortex and hippocampus, alleviating decreased CBF in these two cerebral regions. VEGF-NFMs application improved reduced NAA/Cr ratio, preventing neuronal loss. VEGF-NFMs sticking decreased the number of TUNEL-positive cells in surgical temporal cortex, ameliorated impaired synaptic plasticity, and inhibited the release of pro-inflammatory cytokines and the activation of microglia and astrocytes in surgical temporal cortex and hippocampus. Furthermore, BDNF-TrkB/PI3K/AKT, BDNF-TrkB/ERK and HIF-1a/VEGF/ERK pathways were involved in the treatment of VEGF-NFMs against CCH-induced neuronal injury. These results showed the neuroprotective effects of VEGF-NFMs sticking may initiate from neurovascular repairing followed by inhibition of neuronal apoptosis and neuronal and synaptic damage, eventually leading to the suppression of cognitive dysfunction, which provided theoretical foundation for further clinical transformation of VEGF-NFMs.

Knockout of the odorant receptor co-receptor, orco, impairs feeding, mating and egg-laying behavior in the fall armyworm Spodoptera frugiperda.

The olfactory transduction system of insects is involved in multiple behavioral processes such as foraging, mating, and egg-laying behavior. In the insect olfactory receptor neurons (ORNs), the odorant receptor co-receptor (Orco) is an obligatory component that is required for dimerization with odorant receptors (ORs) to form a ligand-gated ion channel complex. The ORs/Orco heteromeric complex plays a crucial role in insect olfaction. To explore the function of OR-mediated olfaction in the physiological behavior of the fall armyworm, Spodoptera frugiperda, we applied CRISPR/Cas9 genome editing to mutate its Orco gene and constructed a homozygous mutant strain of Orco (Orco-/-) by genetic crosses. Electroantennogram (EAG) analysis showed that the responses of Orco-/- male moths to two universal sex pheromones, Z9-14: Ac and Z7-12: Ac, were abolished. We found that Orco-/- males cannot successfully mate with female moths. An oviposition preference assay confirmed that Orco-/- female moths had a reduced preference for the optimal host plant maize. A larval feeding assay revealed that the time for Orco-/- larvae to locate the food source was significantly longer than in the wild-type. Overall, in the absence of Orco, the OR-dependent olfactory behavior was impaired in both larval and adult stages. Our results confirm that Orco is essential for multiple behavioral processes related to olfaction in the fall armyworm.