Complete chloroplast genomes of eight Delphinium taxa (Ranunculaceae) endemic to Xinjiang, China: insights into genome structure, comparative analysis, and phylogenetic relationships.

BACKGROUND: Delphinium L. represents a taxonomically intricate genus of significant phylogenetic and economic importance in Ranunculaceae. Despite the existence of few chloroplast genome datasets, a comprehensive understanding of genome structures and selective pressures within the genus remains unknown. Furthermore, several taxa in this genus are exclusively found in Xinjiang, China, a region renowned for its distribution and diversity of Chinese and Central Asian Delphinium species. Therefore, investigating the features of chloroplast genomes in this area will provide valuable insights into the evolutionary processes and phylogenetic relationships of the genus. RESULTS: In this study, the eight newly completed chloroplast genomes are examined, ranging in length from 153,979 bp to 154,284 bp. Alongside these, analysing six previously reported taxa re-annotated in Delphinium, 111 unique genes are identified across all samples. Genome structure, distributions of simple sequence repeats and short dispersed repeats, as well as gene content are similar among these Delphinium taxa. Nine hypervariable intergenic spacers and protein coding regions, including ndhF-trnL(TAG), rpl16-intron, rpl33, rps15, rps18, trnK(TTT)-trnQ(TTG), trnP(TGG)-psaJ, trnT(GGT)-psbD and ycf1, are identified among 13 perennial Delphinium. Selective pressure and codon usage bias of all the plastid genes are performed within 14 Delphinium taxa. Phylogenetic analysis based on 14 Delphinium plastomes, alongside two Aconitum (Ranunculaceae) species serving as outgroup taxa, reveals the monophyletic nature of Delphinium. Our findings further discern Delphinium into two distinct clades: perennial species (clade I) and annual species (clade II). In addition, compared with the nrDNA ITS topology, cytological data and morphological characters, D. mollifolium and D. maackianum showed potential involvement in hybridization or polyploidization processes. Excluding these two species, the perennial Delphinium (clade I) exhibits a stronger consistency with the morphology-based system that utilized seed morphology. CONCLUSION: This study represents the first comprehensive analysis of plastomic variations among Delphinium taxa, based on the examination of 14 complete plastomes. The chloroplast genome structure of Delphinium is similar to other angiosperms and possesses the typical quadripartite structure with the conserved genome arrangement and gene features. In addition, the variation of non-coding regions is larger than coding regions of the chloroplast genome. Through DNA sequence divergence across Delphinium plastomes and subsequent phylogenomic analyses ndhF-trnL(TAG) and ycf1 are identified as promising molecular markers. These highly variable loci held significant potential for future phylogenetic and phylogeographic studies on Delphinium. Our phylogenomic analyses based on the whole plastomes, concatenation of 132 unique intergenic spacer regions, concatenation of 77 unique protein-coding genes and nrDNA ITS, all support the monophyly of Delphinium and perennial taxa clusters together into one clade within this genus. These findings provide crucial data for systematic, phylogenomic and evolutionary research in the genus for future studies.

Microfluidic Impedance Cytometry Enabled One-Step Sample Preparation for Efficient Single-Cell Mass Spectrometry.

Single-cell mass spectrometry (MS) is significant in biochemical analysis and holds great potential in biomedical applications. Efficient sample preparation like sorting (i.e., separating target cells from the mixed population) and desalting (i.e., moving the cells off non-volatile salt solution) is urgently required in single-cell MS. However, traditional sample preparation methods suffer from complicated operation with various apparatus, or insufficient performance. Herein, a one-step sample preparation strategy by leveraging label-free impedance flow cytometry (IFC) based microfluidics is proposed. Specifically, the IFC framework to characterize and sort single-cells is adopted. Simultaneously with sorting, the target cell is transferred from the local high-salinity buffer to the MS-compatible solution. In this way, one-step sorting and desalting are achieved and the collected cells can be directly fed for MS analysis. A high sorting efficiency (>99%), cancer cell purity (≈87%), and desalting efficiency (>99%), and the whole workflow of impedance-based separation and MS analysis of normal cells (MCF-10A) and cancer cells (MDA-MB-468) are verified. As a standalone sample preparation module, the microfluidic chip is compatible with a variety of MS analysis methods, and envisioned to provide a new paradigm in efficient MS sample preparation, and further in multi-modal (i.e., electrical and metabolic) characterization of single-cells.

Floating-Electrode-Enabled Impedance Cytometry for Single-Cell 3D Localization.

As a label-free, low-cost, and noninvasive tool, impedance measurement has been widely used in single-cell characterization analysis. However, due to the tiny volume of cells, the uncertainty of the spatial position in the microchannel will bring measurement errors in single-cell electrical parameters. To overcome the issue, we designed a novel microdevice configured with a coplanar differential electrode structure to accurately resolve the spatial position of single cells without constraining techniques such as additional sheath fluids or narrow microchannels. The device precisely localizes single cells by measuring the induced current generated by the combined action of the floating electrode and the differential electrodes when single cells flow through the electrode-sensing area. The device was experimentally validated by measuring 6 µm yeast cells and 10 µm particles, achieving spatial localization with a resolution down to 2.1 µm (about 5.3% of the channel width) in lateral direction and 1.2 µm (about 5.9% of the channel height) in the vertical direction at a flow rate of 1.2 µL/min. In addition, by comparing measurement of yeast cells and particles, it was demonstrated that the device not only localizes the single cells or particles but also simultaneously characterizes their status properties such as velocity and size. The device offers a competitive electrode configuration in impedance cytometry with the advantages of simple structure, low cost, and high throughput, promising cell localization and thus electrical characterization.

Efficient Sample Preparation System for Multi-Omics Analysis via Single Cell Mass Spectrometry.

Mass spectrometry (MS) has become a powerful tool for metabolome, lipidome, and proteome analyses. The efficient analysis of multi-omics in single cells, however, is still challenging in the manipulation of single cells and lack of in-fly cellular digestion and extraction approaches. Here, we present a streamlined strategy for highly efficient and automatic single-cell multi-omics analysis by MS. We developed a 10-pL-level microwell chip for housing individual single cells, whose proteins were found to be digested in 5 min, which is 144 times shorter than traditional bulk digestion. Besides, an automated picoliter extraction system was developed for sampling of metabolites, phospholipids, and proteins in tandem from the same single cell. Also, 2 min MS2 spectra were obtained from 700 pL solution of a single cell sample. In addition, 1391 proteins, phospholipids, and metabolites were detected from one single cell within 10 min. We further analyzed cells digested from cancer tissue samples, achieving up to 40% increase in cell classification accuracy using multi-omics analysis in comparison with single-omics analysis. This automated single-cell MS strategy is highly efficient in analyzing multi-omics information for investigation of cell heterogeneity and phenotyping for biomedical applications.

Impedance-Based Multimodal Electrical-Mechanical Intrinsic Flow Cytometry.

Reflecting various physiological states and phenotypes of single cells, intrinsic biophysical characteristics (e.g., mechanical and electrical properties) are reliable and important, label-free biomarkers for characterizing single cells. However, single-modal mechanical or electrical properties alone are not specific enough to characterize single cells accurately, and it has been long and challenging to couple the conventionally image-based mechanical characterization and impedance-based electrical characterization. In this work, the spatial-temporal characteristics of impedance sensing signal are leveraged, and an impedance-based multimodal electrical-mechanical flow cytometry framework for on-the-fly high-dimensional intrinsic measurement is proposed, that is, Young's modulus E, fluidity ß, radius r, cytoplasm conductivity σi , and specific membrane capacitance Csm , of single cells. With multimodal high-dimensional characterization, the electrical-mechanical flow cytometry can better reveal the difference in cell types, demonstrated by the experimental results with three types of cancer cells (HepG2, MCF-7, and MDA-MB-468) with 93.4% classification accuracy and pharmacological perturbations of the cytoskeleton (fixed and Cytochalasin B treated cells) with 95.1% classification accuracy. It is envisioned that multimodal electrical-mechanical flow cytometry provides a new perspective for accurate label-free single-cell intrinsic characterization.

Sensitive marker for evaluation of hypertensive heart disease: extracellular volume and myocardial strain.

BACKGROUND: Evaluation of tissue fibrosis and myocardial hypertrophy in left ventricular (LV) remodeling is the basis of post-treatment evaluation of hypertensive heart disease (HHD). Extracellular volume (ECV) and myocardial strain parameters can indirectly reflect the changes of both. Our objective was to analyze the characteristics of ECV and strain parameters in LV myocardium of HHD with varying degrees of systolic dysfunction, and to explore the changes of both after treatment for hypertension. METHODS: A total of 62 HHD patients were divided into 3 groups according to ejection fraction (EF < 30, 30%≦EF < 50%, EF≧50%). Twenty-one of these patients underwent cardiac magnetic resonance (CMR) reexamination more than six months after receiving antihypertensive medication. The initial T1 time and post-enhancement T1 time of each segment were measured, and the ECV was calculated. Radial strain (RS), circumferential strain (CS) and longitudinal strain (LS) of LV were measured by cvi42 software, and the differences in CMR parameters between different groups and before and after treatment were compared. RESULTS: ①The mean, basal and middle ECV value of HHD groups with different EF were all higher than that of the control group (P < 0.05), but the difference between HHD groups was not statistically significant. ②With the decrease of EF, the absolute value of both the global or local strain decreased. Strain is related to LVMI and ECV. ③In general, ECV, global RS (GRS) and global CS (GCS) improved after treatment, but the improvement of LS impairment in HHD patients is difficult. CONCLUSIONS: ECV and myocardial strain parameters are more sensitive to myocardial abnormalities, and ECV, GRS and GCS are more sensitive to treatment. However it is difficult to improve longitudinal strain impairment in HHD patients. ECV and myocardial strain parameters can be used as good makers for long-term monitoring of the efficacy of HHD patients.

The study on size dependent dipole-dipole interaction in the self-assembly of twisting nanoribbons with circular polarization activation.

The mechanism of self-assembling process of inorganic nanoparticle (NPs) is still an open question due to the various and non-additive interactions between NPs. Kotov et al reported that the semiconductor NPs can be self-assembled by external activation such as irradiation. In this paper, the twisted CdTe nanoribbons were successfully assembled with circular polar light activation based on the chiral selective resonance absorption. The effect of NP size on the morphology of assemblies under circular polar light irradiation is discussed by introducing a new mechanism of photooxidation induced dipole moment which decreases with increasing sizes of the NPs because of the change of band offsets at the CdS/CdTe interface. Moreover, we find that the competition between the dipole-dipole interaction and electrostatic repulsion can be modulated by the size of the NPs and the concentration of dispersion, which are the key points to produce the chiral twisted nanoribbons.

Is it Suitable for a Journal to Bid for Publishing a Review That is Likely to be Highly Cited?

By following a recently published paper entitled "The effect of publishing a highly cited paper on a journal's impact factor: a case study of the Review of Particle Physics" in Learned Publishing, we argue that it is not suitable for journals to bid for the right to publish a review that is likely to be highly cited. A few suggestions are also provided to deal with the special case of the Review of Particle Physics phenomenon.

The Modulatory Effects of Curcumin on the Gut Microbiota: A Potential Strategy for Disease Treatment and Health Promotion.

Curcumin (CUR) is a lipophilic natural polyphenol that can be isolated from the rhizome of turmeric. Studies have proposed that CUR possesses a variety of biological activities. Due to its anti-inflammatory and antioxidant properties, CUR shows promise in the treatment of inflammatory bowel disease, while its anti-obesity effects make it a potential therapeutic agent in the management of obesity. In addition, curcumin's ability to prevent atherosclerosis and its cardiovascular benefits further expand its potential application in the treatment of cardiovascular disease. Nevertheless, owing to the limited bioavailability of CUR, it is difficult to validate its specific mechanism of action in the treatment of diseases. However, the restricted bioavailability of CUR makes it challenging to confirm its precise mode of action in disease treatment. Recent research indicates that the oral intake of curcumin may lead to elevated levels of residual curcumin in the gastrointestinal system, hinting at curcumin's potential to directly influence gut microbiota. Furthermore, the ecological dysregulation of the gut microbiota has been shown to be critical in the pathogenesis of human diseases. This review summarizes the impact of gut dysbiosis on host health and the various ways in which curcumin modulates dysbiosis and ameliorates various diseases caused by it through the administration of curcumin.

Inductive Paper-Based Flexible Contact Force Sensor Utilizing Natural Micro-Nanostructures of Paper: Simplicity, Economy, and Eco-Friendliness.

Inductive contact force sensors, known for their high precision and anti-interference capabilities, hold significant potential applications in fields such as wearable and medical monitoring devices. Most of the current research on inductive contact force sensors employed novel nanomaterials as sensitive elements to enhance their sensitivity and other performance characteristics. However, sensors developed through such methods typically involve complex preparation processes, high costs, and difficulty in biodegradation, which limit their further development. This article introduces a new flexible inductive contact force sensor using paper as a sensitive element. Paper inherently possesses micro- and nanostructures on its surface and interior, enabling it to sensitively convert changes in contact force into changes in displacement, making it suitable for use as the sensor's sensitive element. Additionally, the advantages of paper also include its great flexibility, low cost, wide availability, and biodegradability. Performance testing on this flexible sensor showed good repeatability, hysteresis, sensitivity, and consistency. When used in experiments for monitoring human motion and respiration, this sensor also exhibited great detection performance. The proposed inductive paper-based flexible contact force sensor, with its simple structure, easy manufacturing process, cost-effectiveness, eco-friendliness, and good sensing performance, provides new insights into research for contact force sensors.

Highly Sensitive Paper-Based Force Sensors with Natural Micro-Nanostructure Sensitive Element.

Flexible paper-based force sensors have garnered significant attention for their important potential applications in healthcare wearables, portable electronics, etc. However, most studies have only used paper as the flexible substrate for sensors, not fully exploiting the potential of paper's micro-nanostructure for sensing. This article proposes a novel approach where paper serves both as the sensitive element and the flexible substrate of force sensors. Under external mechanical forces, the micro-nanostructure of the conductive-treated paper will change, leading to significant changes in the related electrical output and thus enabling sensing. To demonstrate the feasibility and universality of this new method, the article takes paper-based capacitive pressure sensors and paper-based resistive strain sensors as examples, detailing their fabrication processes, constructing sensing principle models based on the micro-nanostructure of paper materials, and testing their main sensing performance. For the capacitive paper-based pressure sensor, it achieves a high sensitivity of 1.623 kPa-1, a fast response time of 240 ms, and a minimum pressure resolution of 4.1 Pa. As for the resistive paper-based strain sensor, it achieves a high sensitivity of 72 and a fast response time of 300 ms. The proposed new method offers advantages such as high sensitivity, simplicity in the fabrication process, environmental friendliness, and cost-effectiveness, providing new insights into the research of flexible force sensors.

Capillarity Enabled Large-Array Liquid Metal Electrodes for Compact and High-Throughput Dielectrophoretic Microfluidics.

Dielectrophoresis (DEP) particle separation has label-free, well-controllable, and low-damage merits. Sidewall microelectrodes made of liquid metal alloy (LMA) inherits the additional advantage of thick electrodes to generate impactful DEP force. However, existing LMA electrode-based devices lack the ability to integrate large-array electrodes in a compact footprint, severely limiting flow rate and thus throughput. Herein, a facile and versatile method is proposed to integrate high-density thick LMA electrodes in microfluidic devices, taking advantage of the passive control ability of capillary burst valves (CBVs). CBVs with carefully designed burst pressures are co-designed in microfluidic channels, allowing self-assembly of LMA electrode array through simple hand-push injection. The arrayed electrode configuration brings the accumulative DEP deflection effect. Specifically, The fabricated 5000 pairs of sidewall electrodes in a compact chip are demonstrted to achieve ten times higher throughput in DEP deflection. The 5000-electrode-pair device is applied to successfully separate four mixed samples, including human peripheral blood mononuclear cells and A549 cells with the flow rate of 70 µL min-1. It is envisioned that this work can greatly facilitate LMA electrode array fabrication and offer a robust and versatile platform for DEP separation applications.

Comprehensive analysis of glycolysis mediated pattern clusters and immune infiltration characterization of tumor microenvironment in triple-negative breast cancer.

Background: The involvement of glycolysis in carcinogenesis and the tumor microenvironment is being increasingly supported by the available data. The aim of this work was to develop a triple-negative breast cancer predictive model based on glycolysis. Methods: Glycolysis mediated pattern clusters were created using the R "ConsensusClusterPlus" package. The variations in the tumor microenvironment between the pattern clusters were examined using the R "GSVA", "ESTIMATE", and "CIBERSORT" package. The risk score and nomogram were established to assess clinical outcomes of patients. Results: Substantial differences were observed in the immunological landscape between the glycolysis-mediated pattern clusters. When it came to predicting survival and immunotherapy response, the developed risk score showed promising predictive power. Nomogram was designed to be used in therapeutic settings due to its remarkable predictive accuracy. Conclusions: The immune microenvironment varied among cases of triple-negative breast cancer. The nomogram and the risk score based on glycolysis could both be used to help create more effective treatments.

Enhanced E6AP-mediated ubiquitination of ENO1 via LINC00663 contributes to radiosensitivity of breast cancer by regulating mitochondrial homeostasis.

Radiotherapy has shown measurable efficacy in breast cancer (BC). Elucidating the mechanisms and developing effective strategies against resistance, which is a major challenge, is crucial. Mitochondria, which regulate homeostasis of the redox environment, have emerged as a radiotherapeutic target. However, the mechanism via which mitochondria are controlled under radiation remains elusive. Here, we identified alpha-enolase (ENO1), as a prognostic marker for the efficacy of BC radiotherapy. ENO1 enhances radio-therapeutic resistance in BC via reducing the production of reactive oxygen species (ROS) and apoptosis in vitro and in vivo through modulation of mitochondrial homeostasis. Moreover, LINC00663 was identified as an upstream regulator of ENO1, which regulates radiotherapeutic sensitivity by downregulating ENO1 expression in BC cells. LINC00663 regulates ENO1 protein stability by enhancing the E6AP-mediated ubiquitin-proteasome pathway. In BC patients, LINC00663 expression is negatively correlated with ENO1 expression. Among patients treated with IR, those who did not respond to radiotherapy expressed lower levels of LINC00663 than those sensitive to radiotherapy. Our work established LINC00663/ENO1 critical to regulate IR-resistance in BC. Inhibition of ENO1 with a specific inhibitor or supplement of LINC00663 could be potential sensitizing therapeutic strategies for BC.

Discovery of a heterotrophic aerobic denitrification Pseudomonas sp. G16 and its unconventional nitrogen metabolic pathway.

From the aerobic pond of the farm, the Pseudomonas sp. G16 was screened and isolated, which was confirmed to exhibit heterotrophic nitrification and aerobic denitrification. The removal rates of Ammonia (100 mg/L), nitrate (120 mg/L), and nitrite (100 mg/L) by the strain were 94.13%, 92.62%, and 85.67%, and the nitrogen metabolism pathway of strain G16 was analyzed by whole genome sequencing combined with its nitrification-denitrification intermediate products, it was found that the strain had independent nitrification-denitrification ability and no nitrite accumulation. Under the conditions of carbon source of sodium succinate hexahydrate, C/N ratio of 15, pH of 7.5, temperature of 15 °C, and DO of 210 rpm, strain G16 showed excellent denitrification performance. Strain G16 was prepared into biochar-based immobilized bacterial particles, which successfully improved its nitrogen removal efficiency and stability. Therefore, the application of strain G16 in the field of real wastewater treatment has very necessary research value.

Performance-enhanced clogging-free viscous sheath constriction impedance flow cytometry.

As a label-free and high-throughput single cell analysis platform, impedance flow cytometry (IFC) suffers from clogging caused by a narrow microchannel as mechanical constriction (MC). Current sheath constriction (SC) solutions lack systematic evaluation of the performance and proper guidelines for the sheath fluid. Herein, we hypothesize that the viscosity of the non-conductive liquid is the key to the performance of SC, and propose to employ non-conductive viscous sheath flow in SC to unlock the tradeoff between sensitivity and throughput, while ensuring measurement accuracy. By placing MC and SC in series in the same microfluidic chip, we established an evaluation platform to prove the hypothesis. Through modeling analysis and experiments, we confirmed the accuracy (error < 1.60% ± 4.71%) of SC w.r.t. MC, and demonstrated that viscous non-conductive PEG solution achieved an improved sensitivity (7.92×) and signal-to-noise ratio (1.42×) in impedance measurement, with the accuracy maintained and free of clogging. Viscous SC IFC also shows satisfactory ability to distinguish different types of cancer cells and different subtypes of human breast cancer cells. It is envisioned that viscous SC IFC paves the way for IFC to be really usable in practice with clogging-free, accurate, and sensitive performance.

Gut macrobiotic and its metabolic pathways modulate cardiovascular disease.

Thousands of microorganisms reside in the human gut, and extensive research has demonstrated the crucial role of the gut microbiota in overall health and maintaining homeostasis. The disruption of microbial populations, known as dysbiosis, can impair the host's metabolism and contribute to the development of various diseases, including cardiovascular disease (CVD). Furthermore, a growing body of evidence indicates that metabolites produced by the gut microbiota play a significant role in the pathogenesis of cardiovascular disease. These bioactive metabolites, such as short-chain fatty acids (SCFAs), trimethylamine (TMA), trimethylamine N-oxide (TMAO), bile acids (BAs), and lipopolysaccharides (LPS), are implicated in conditions such as hypertension and atherosclerosis. These metabolites impact cardiovascular function through various pathways, such as altering the composition of the gut microbiota and activating specific signaling pathways. Targeting the gut microbiota and their metabolic pathways represents a promising approach for the prevention and treatment of cardiovascular diseases. Intervention strategies, such as probiotic drug delivery and fecal transplantation, can selectively modify the composition of the gut microbiota and enhance its beneficial metabolic functions, ultimately leading to improved cardiovascular outcomes. These interventions hold the potential to reshape the gut microbial community and restore its balance, thereby promoting cardiovascular health. Harnessing the potential of these microbial metabolites through targeted interventions offers a novel avenue for tackling cardiovascular health issues. This manuscript provides an in-depth review of the recent advances in gut microbiota research and its impact on cardiovascular health and offers a promising avenue for tackling cardiovascular health issues through gut microbiome-targeted therapies.

Evaluating the Accuracy of Impedance Flow Cytometry with Cell-Sized Liposomes.

Electrical properties of single cells are important label-free biomarkers of disease and immunity. At present, impedance flow cytometry (IFC) provides important means for high throughput characterization of single-cell electrical properties. However, the accuracy of the spherical single-shell electrical model widely used in IFC has not been well evaluated due to the lack of reliable and reproducible single-shell model particles with true-value electrical parameters as benchmarks. Herein, a method is proposed to evaluate the accuracy of the single-cell electrical model with cell-sized unilamellar liposomes synthesized through double emulsion droplet microfluidics. The influence of three key dimension parameters (i.e., the measurement channel width w, height h, and electrode gap g) in the single-cell electrical model were evaluated through experiment. It was found that the relative error of the electrical intrinsic parameters measured by IFC is less than 10% when the size of the sensing zone is close to the measured particles. It further reveals that h has the greatest influence on the measurement accuracy, and the maximum relative error can reach ∼30%. Error caused by g is slightly larger than w. This provides a solid guideline for the design of IFC measurement system. It is envisioned that this method can advance further improvement of IFC and accurate electrical characterization of single cells.

Non-Invasive and Minute-Frequency 3D Tomographic Imaging Enabling Long-Term Spatiotemporal Observation of Single Cell Fate.

Non-invasive and rapid imaging technique at subcellular resolution is significantly important for multiple biological applications such as cell fate study. Label-free refractive-index (RI)-based 3D tomographic imaging constitutes an excellent candidate for 3D imaging of cellular structures, but its full potential in long-term spatiotemporal cell fate observation is locked due to the lack of an efficient integrated system. Here, a long-term 3D RI imaging system incorporating a cutting-edge white light diffraction phase microscopy module with spatiotemporal stability, and an acoustofluidic device to roll and culture single cells in a customized live cell culture chamber is reported. Using this system, 3D RI imaging experiments are conducted for 250 cells and demonstrate efficient cell identification with high accuracy. Importantly, long-term and frequency-on-demand 3D RI imaging of K562 and MCF-7 cancer cells reveal different characteristics during normal cell growth, drug-induced cell apoptosis, and necrosis of drug-treated cells. Overall, it is believed that the proposed 3D tomographic imaging technique opens up a new avenue for visualizing intracellular structures and will find many applications such as disease diagnosis and nanomedicine.

Induced formation of primordial germ cells from zebrafish blastomeres by germplasm factors.

The combination of genome editing and primordial germ cell (PGC) transplantation has enormous significance in the study of developmental biology and genetic breeding, despite its low efficiency due to limited number of donor PGCs. Here, we employ a combination of germplasm factors to convert blastoderm cells into induced PGCs (iPGCs) in zebrafish and obtain functional gametes either through iPGC transplantation or via the single blastomere overexpression of germplasm factors. Zebrafish-derived germplasm factors convert blastula cells of Gobiocypris rarus into iPGCs, and Gobiocypris rarus spermatozoa can be produced by iPGC-transplanted zebrafish. Moreover, the combination of genome knock-in and iPGC transplantation perfectly resolves the contradiction between high knock-in efficiency and early lethality during embryonic stages and greatly improves the efficiency of genome knock-in. Together, we present an efficient method for generating PGCs in a teleost, a technique that will have a strong impact in basic research and aquaculture.