Biology subjects

Wu, Y.

Publications and source records attributed to Wu, Y..

At least 379 records · Page 21Linked to original sources

bioRxiv2025

ST-FFPE-mIF: Integrating Spatial Transcriptomics and Multiplex Immunofluorescence in Formalin-Fixed Paraffin-Embedded Tissues Using Stereo-seq

Stereo-seq is the state-of-the-art technique, which allows capturing of spatial transcriptomic data at sub-cellular resolution and in large scale of view. Stereo-seq FFPE technique is recently launched with the capability to obtain spatially resolved gene expression in FFPE tissue sections. However, the omic information is limited to transcriptome. To delineate spatial molecular hierarchy from transcriptome to phenome, we present the protocol that implements nine-plex immunofluorescence staining with detection of spatial transcriptomics in one FFPE tissue section (short for ST-FFPE-mIF). This protocol details the modified biochemistry procedure and also provides analytical framework to achieve single-cell resolution through nuclear or membrane staining-based cell segmentation. We demonstrated the application of ST-FFPE-mIF in colorectal cancer sample to illustrate how ST-FFPE-mIF facilitates a robust analysis of spatially resolved multi-omic data at the single-cell level.

biochemistry↗

bioRxiv2025

Development of a novel murine model of in-stent neoatherosclerosis

ObjectiveIn-stent neoatherosclerosis is a phenomenon of percutaneous coronary intervention with stenting. Whilst similar to de novo atherosclerosis, it develops rapidly over 1-5 years rather than over a lifetime. No preclinical small animal models exist that allow full elucidation of neoatherosclerosis biology and future treatments. The aim of this study was to establish and validate a novel murine model of in-stent neoatherosclerosis. Approach and ResultsMurine stainless-steel stents (2.5 x 0.7 mm) were deployed into donor descending aortas of atherosclerosis-prone apolipoprotein (Apo)e-/- mice, then carotid-interposition grafted into Apoe-/- recipients. Mice (n=6-8/group) received chow or a high cholesterol diet (HCD) for 7- or 28-days post-surgery. Multimodal intravascular imaging, simultaneously combining optical coherence tomography (OCT, plaque burden) and fluorescence for indocyanine green (ICG, plaque instability), visualized in-stent neoatherosclerosis across the entire length of the stented site. Histological analyses revealed that stented vessels from mice fed HCD had neointimas with prominent lipid cores and an elevated CD68+ macrophage content, similar to human neoatherosclerosis. Mice fed chow post-stenting had distinctly different neointimas that were smooth muscle cell rich, resembling neointimal hyperplasia. Consistent with this, flow cytometry revealed a higher content of monocytes/macrophages and dendritic cells in stented aortas from mice fed HCD than in non-stented aortas. ConclusionWe have developed and validated the first murine model that replicates the unique characteristics of human in-stent neoatherosclerosis. This project has implications for exploring the mechanisms that promote neoatherosclerosis and testing targeted new therapies. RESEARCH PERSPECTIVEO_ST_ABSWhat Is New?C_ST_ABSO_LIWe have developed and validated a novel murine model of in-stent neoatherosclerosis, presenting a new platform that will facilitate the discovery of novel mechanistic targets of in-stent neoatherosclerosis and preventative therapies. C_LIO_LIThis model develops lesions with a similar morphology to human in-stent neoatherosclerosis and distinct to in-stent neointimal hyperplasia, with higher extracellular lipid and macrophage content and proportionately less smooth muscle cells. C_LIO_LIWe show a first-time visualization of murine in-stent neoatherosclerosis using bimodal intravascular imaging with simultaneous capture of structural information (optical coherence tomography, plaque burden) and the distribution of areas of plaque instability (high-sensitivity fluorescence, indocyanine green) within the plaque. C_LI What new question does this study raise?O_LIHow can the utility of this novel model be maximized as a platform for discovering novel agents that prevent in-stent neoatherosclerosis? C_LI What question should be addressed next?O_LIAre there unique mechanisms of in-stent neoatherosclerosis, distinct to de novo atherosclerosis, that can be specifically targeted to prevent disease and ultimately increase stent performance? C_LI

cell biology↗

bioRxiv2025

Computational Modeling of Ternary Complex Formation and Targeted Protein Degradation Mediated by Molecular Glues

The rational design of molecular glue degraders is challenging because glue-mediated protein degradation depends on a complex interplay of molecular mechanisms: an effective molecular glue must simultaneously bring together two distinct proteins to form an interface that is both stable and accessible for ubiquitination by E3 ligases. We present GlueMap, a computational platform for discovering and optimizing molecular glues (MGs) through integrative structural and pharmacodynamic modeling. GlueMap combines molecular dynamics (MD) simulations and machine learning (ML) to model targeted protein degradation (TPD) mediated by molecular glues. We validated GlueMap by applying it to two case studies: DDB1-dependent degradation of CDK12 and CRBN-dependent degradation of GSPT1. In the DDB1-CDK12 system, GlueMap revealed strong correlations between ternary-complex stability, quantified through rigorous free-energy calculations, and experimental degradation efficiency. Notable outliers underscored the importance of incorporating conformational dynamics within assembled Cullin-RING ligase complexes. Using a supervised variational autoencoder combined with attention-based regression, GlueMap achieved high predictive accuracy for degradation potency from structural features. For CRBN-GSPT1, GlueMap reproduced known ternary-complex crystal structures and identified previously unrecognized protein-protein interactions (PPIs) that may guide the design of next-generation glues. Hierarchical virtual screening of more than 12 billion compounds successfully recovered known degraders and uncovered nine new potent MGs with DC50 values below 50 nM. Across both systems, GlueMap outperformed Boltz-1 and Boltz-2, state-of-the-art structure-prediction models that failed to reproduce the CDK12-DDB1 crystal structure and captured substantially less PPI diversity. By integrating structural, thermodynamic, and ubiquitination metrics, GlueMap establishes a multi-criteria framework for rational molecular glue design. These results demonstrate that GlueMap provides both mechanistic insight and practical acceleration of molecular glue discovery and optimization across diverse therapeutic targets.

bioinformatics↗

bioRxiv2025

Melatonin attenuates kidney injury by alleviating lysosomal damage in diabetic kidney disease

Proteinuria-induced damage to renal tubular epithelial cells is one of the main causes of diabetic kidney disease (DKD), and the clearance of overloaded albumin by lysosomes is crucial for maintaining the homeostasis of renal tubular epithelial cells. Therefore, lysosomal damage is closely related to the pathogenesis of DKD, but effective prevention and treatment measures are still lacking. Melatonin (MLT) is secreted by the pineal gland and can not only regulate circadian rhythms but also maintain lysosomal homeostasis. In this study, we demonstrate the presence of significant lysosomal damage in renal tubules of DKD patients, which causes autophagy impairment and a concomitant oxidative stress imbalance; however, MLT can upregulate transcription factor EB (TFEB) to improve lysosomal damage and restore the biosynthesis in this organelle. Mechanistically, MLT may protect lysosomes via the upregulation of TFEB and the miR-205-5p-LRP-1 pathway in renal tubules, thus improving autophagy dysfunction and oxidative imbalance in DKD.

pathology↗

bioRxiv2024

Single-cell analysis of the epigenome and 3D chromatin architecture in the human retina

Most genetic risk variants linked to ocular diseases are non-protein coding and presumably contribute to disease through dysregulation of gene expression, however, deeper understanding of their mechanisms of action has been impeded by an incomplete annotation of the transcriptional regulatory elements across different retinal cell types. To address this knowledge gap, we carried out single-cell multiomics assays to investigate gene expression, chromatin accessibility, DNA methylome and 3D chromatin architecture in human retina, macula, and retinal pigment epithelium (RPE)/choroid. We identified 420,824 unique candidate regulatory elements and characterized their chromatin states in 23 sub-classes of retinal cells. Comparative analysis of chromatin landscapes between human and mouse retina cells further revealed both evolutionarily conserved and divergent retinal gene-regulatory programs. Leveraging the rapid advancements in deep-learning techniques, we developed sequence-based predictors to interpret non-coding risk variants of retina diseases. Our study establishes retina-wide, single-cell transcriptome, epigenome, and 3D genome atlases, and provides a resource for studying the gene regulatory programs of the human retina and relevant diseases.

genomics↗

bioRxiv2024

PSGL-1 excludes HIV Env from virion surface through spatial hindrance involving structural folding of the decameric repeats (DR)

P-selectin glycoprotein ligand-1 (PSGL-1), a mucin-like surface glycoprotein, is primarily expressed on lymphoid and myeloid cells. PSGL-1 has recently been identified as an HIV restriction factor, blocking HIV infectivity mainly through virion incorporation that sterically hinders virion attachment to target cells. PSGL-1 also inhibits HIV Env incorporation into virions. However, the molecular mechanisms of PSGL-1-mediated Env exclusion remained unclear. Here, we investigated the role of PSGL-1s extracellular (EC) and intracellular (IC) domains in Env exclusion. We demonstrate that both EC and IC are important for Env exclusion; when EC was deleted, PSGL-1 completely lost its ability to inhibit Env incorporation, whereas when IC was deleted, PSGL-1 partially lost this activity. In addition, when the decameric repeats (DR) were deleted from EC, PSGL-1 also lost its ability to inhibit Env incorporation. Sequential DR deletion mutagenesis further demonstrated that a minimum of 9 DRs is necessary for Env exclusion. Molecular modeling of the DR structure revealed that PSGL-1 mutants with 7 or fewer DRs pose as an extended "rod-like" structure, whereas those with 9 or more DRs collapse into a "coil-like" structure that spatially excludes Env. Our studies suggest a model in which Env exclusion involves Gag-mediated PSGL-1 targeting to the virion assembly site where DR-mediated spatial exclusion blocks Env incorporation.

microbiology↗

bioRxiv2024

The Restoration of Social Defects in Schizophrenic Mice by Plant Exposure

Schizophrenia is a serious psychotic disorder caused by both individuals genetic background and their living environment. However, how the cognitive and negative symptoms can be treated is still a challenge since most anti-psychotic drugs are only effective for positive symptoms. Previous epidemic studies have demonstrated that plant exposure could decrease the risk of schizophrenia and shorten the length of psychiatric hospital admissions for patients. However, it is still unknown whether plant exposure could improve cognition-related defects in schizophrenia, with no related animal studies. In the present study, we first induced a schizophrenia mice model by giving mice long-term (2 weeks) injections of the antagonist of the NMDA receptor: MK801. We then raised the animals in environments containing plants (4 weeks) including Epipremnum aureum and rosemary, and tested their locomotive, anxious and social behaviors. We found that plant exposure did not change the locomotion behaviors of wild-type animals, but significantly reduced the schizophrenia-related social and anxious behaviors in the schizophrenic animals. In addition, we tested the expression of c-Fos in animals exposed to plants after social behavioral testing and found that the deficits in c-Fos expression in both the hippocampus and prefrontal cortex were partially rescued after plant exposure. These results indicate plant exposure will be a new tool to improve the clinical deficits of schizophrenia.

neuroscience↗

bioRxiv2024

Notch signaling maintains a progenitor-like subclass of hepatocellular carcinoma

Hepatocellular carcinomas (HCCs) constitute one of the few cancer indications for which mortality rates continue to rise. While Notch signaling dictates a key progenitor lineage choice during development, its role in HCC has remained controversial. Using therapeutic antibodies targeting Notch ligands and receptors to screen over 40 patient-derived xenograft models, we here identify progenitor-like HCCs that crucially depend on a tumor-intrinsic JAG1-NOTCH2 signal. Inhibiting this signal induces tumor regressions by triggering progenitor-to-hepatocyte differentiation, the same cell fate-switch that Notch controls during development. Transcriptomic analysis places the responsive tumors within the well-characterized progenitor subclass, a poor prognostic group of highly proliferative tumors, providing a diagnostic method to enrich for Notch-dependent HCCs. Furthermore, single-cell RNA sequencing uncovers a heterogeneous population of tumor cells and reveals how Notch inhibition shifts cells from a mixed cholangiocyte-hepatocyte lineage to one resembling mature hepatocytes. Analyzing the underlying transcriptional programs brings molecular detail to this process by showing that Notch inhibition de-represses expression of CEBPA, which enables the activity of HNF4, a hepatocyte lineage factor that is otherwise quiescent. We thus describe a compelling and targetable dependency in a poor-prognosis class of HCCs.

cancer biology↗

bioRxiv2024

Unprecedented female mutation bias in aye-ayes

Every mammal studied to date has been found to have a male mutation bias: male parents transmit more de novo mutations to offspring than female parents, contributing increasingly more mutations with age. Although male-biased mutation has been studied for more than 75 years, its causes are still debated. One obstacle to understanding this pattern is its near universality--without variation in mutation bias, it is difficult to find an underlying cause. Here, we present new data on multiple pedigrees from two primate species: aye-ayes (Daubentonia madagascarensis), a member of the strepsirrhine primates, and olive baboons (Papio anubis). In stark contrast to the pattern found across mammals, we find a much larger effect of maternal age than paternal age on mutation rates in the aye-aye. In addition, older aye-aye mothers transmit substantially more mutations than older fathers. We carry out both computational and experimental validation of our results, contrasting them with results from baboons and other primates using the same methodologies. Further, we analyze a set of DNA repair and replication genes to identify candidate mutations that may be responsible for the change in mutation bias observed in aye-ayes. Our results demonstrate that mutation bias is not an immutable trait, but rather one that can evolve between closely related species. Further work on aye-ayes (and possibly other lemuriform primates) should help to explain the molecular basis for sex-biased mutation.

evolutionary biology↗

bioRxiv2024

Spatial drug asymmetry modulates phenotypic diversity-migration relationships under resistance evolution

At long timescales, resistant phenotypes will emerge and be selected within the bacterial population as an evolutionary response to drug exposure. This phenomenon reduces the efficacy of drug therapies and thus compromises patient health. In spatially heterogeneous drug environments, recent evidence shows that migration can either promote or decelerate the evolution of antibiotic resistance, thereby affecting the rate of resistant phenotype emergence. However, another important quantitative aspect of resistance evolution--bacterial phenotypic diversity--has often been overlooked and remains challenging to investigate in spatially extended systems, both experimentally and clinically. In order to study how diversity is reshaped by migration across space, here we designed a minimal 2-well experimental system with spatial drug asymmetry. One well contained a bacteriostatic drug (Linezolid) at the minimum inhibitory concentration, while the other well served as a sanctuary with just media. We found that the relationship between diversity and migration follows the "Intermediate Disturbance Hypothesis" (IDH), with migration as the disturbance to each well. By varying the selective drug concentrations, we observed that the diversity-migration relationship changes, and IDH can disappear. This behavior was explained by an asymmetry parameter derived from a two-phenotype growth-migration dynamic model. To further validate how different spatial drug asymmetries modulate the diversity-migration relationship through this asymmetry parameter, we applied another bactericidal drug, Ampicillin, and observed similar results. In a more complex scenario involving both Linezolid and Ampicillin, four distinct phenotypes, including cross-resistant variants, emerged. Our asymmetry parameter successfully explained the diversity-migration relationship, with unique diversity dynamics such as multiple peaks appearing from the model. The minimal generalist-specialist framework predicted these unique behaviors through global fitness advantages. Our findings provide experimental support and theoretical explanations for the emergence of phenotypic diversity in clinical settings, such as gut-lung translocation. These insights may pave the way for improved clinical strategies to manage antibiotic resistance evolution.

evolutionary biology↗

bioRxiv2024

Dual roles of IQCG, a novel microtubule nucleation factor rapidly evolving in humans

The centrosome serves as the main microtubule-organizing structure, playing crucial roles in animal development. Uncovering new components and elucidating their functions helps to understand the regulation of cell maintenance and division by the centrosome. In this study, through a combination of genetic analyses and cell biological characterizations, we identify IQ Motif Containing G (IQCG) as a novel microtubule nucleation factor with a dual role involved in both centrosomal microtubule organization and mitotic spindle formation. IQCG interacts with GSK3{beta} via its N2 region, enhancing its centrosomal accumulation in interphase; subsequently, IQCG disperses into the spindle to promote microtubule generation during mitosis and ensure robust cell division. Furthermore, our analyses indicate that the IQCG gene has undergone accelerated evolution within the human lineage, with positive selection acting on the variable N2 region. This adaptation may confer advantages to human cells by augmenting central regulatory control over cellular activities through the centrosome. Our findings offer novel insights into the functional mechanisms and evolutionary significance of the human centrosome associated with human evolution. Summary blurbThe cellular functions of the novel microtubule nucleation factor IQCG, which enhances centrosomal microtubule organization and cell division robustness, are implicated in human evolution.

cell biology↗

bioRxiv2024

Enhancing RNA Delivery: Practical Insights into NeoLNP Transfection Reagent

The rapid advancement of RNA-based therapeutics, particularly in the wake of COVID-19 vaccine success, has prompted significant research into optimizing RNA delivery mechanisms. This study evaluates the NeoLNP RNA Transfection Kit developed by Scindy Pharmaceutical, which utilizes lipid nanoparticles (LNPs) for efficient RNA encapsulation and delivery. We systematically investigate various parameters affecting transfection efficiency, including RNA concentration, RNA/LNP volume ratios, mixing techniques, LNP stability, and culture media. Our results demonstrate that the optimal RNA concentration for transfection efficiency is around 40-60 ng/{micro}L, with a 1:0.75-1:1 RNA-to-LNP ratio yielding the highest protein expression. Additionally, we find that gentle mixing techniques outperform harsher methods, and the stability of LNP-RNA complexes significantly influences transfection outcomes. This research provides practical guidelines for enhancing RNA transfection efficiency, paving the way for more effective RNA therapeutics.

molecular biology↗

bioRxiv2024

Cells Prioritize the Regulation of Cell Mass Density

A cells global physical state is characterized by its volume and dry mass. The ratio of cell mass to volume is the cell mass density (CMD), which is also a measure of macromolecular crowding and concentrations of all proteins. Using the Fluorescence eXclusion method (FXm) and Quantitative Phase Microscopy (QPM), we investigate CMD dynamics after exposure to sudden media osmolarity change. We find that while the cell volume and mass exhibit complex behavior after osmotic shock, CMD follows a straightforward monotonic recovery in 48 hours. The recovery is cell-cycle independent and relies on a coordinated adjustment of protein synthesis and volume growth rates. Surprisingly, we find that the protein synthesis rate decreases when CMD increases. This result is explained by CMD-dependent nucleoplasm-cytoplasm transport, which serves as negative regulatory feedback on CMD. The Na+/H+ exchanger NHE plays a role in regulating CMD by affecting both protein synthesis and volume change. Taken together, we reveal that cells possess a robust control system that actively regulates CMD during environmental change.

biophysics↗

bioRxiv2024

SOX2-induced IL-1alpha-mediated immune suppression drives epithelial dysplasia malignant transformation

Squamous cell carcinomas (SCC) are often preceded by potentially malignant precursor lesions, most of which remain benign. The terminal exhaustion phenotypes of effector T-cells and the accumulation of myeloid-derived suppressor cells (MDSC) have been thoroughly characterized in established SCC. However, it is unclear what precancerous lesions harbor a bona fide high risk for malignant transformation and how precancerous epithelial dysplasia drives the immune system to the point of no return. Here we show that expression of SRY-box transcription factor 2 (SOX2) in precancerous lesions imparts an irreversible risk that recruits suppressive myeloid cells by promoting the release of CCL2. We developed a unique genetically engineered mouse model (GEMM) to recapitulate the malignant transformation of epithelial dysplasia to SCC in the oral mucosa with high histologic and phenotypic fidelity. Using a combination of longitudinal human specimens and the Sox2-GEMM, we found that the myeloid cells in precancerous epithelial dysplasia exhibit a distinctive dichotomous profile featuring high levels of IL-1-SLC2A1 and low levels of type-I interferon (IFN-I) signatures, which occurs before SCC emerges histologically. Brief priming of myeloid cells with IL-1 desensitizes them to IFN-I agonists and makes myeloid-derived suppressor cells (MDSC) even more suppressive of T-cell activation. Mechanistically, IL-1 activation represses the expression of DHHC3/7 enzymes, which are responsible for the palmitoylation of stimulator of interferon genes (STING). Early blockade of IL1 signaling using pharmacologic and genetic approaches similarly reduces MDSC and SLC2A1high myeloid cells, suppresses epithelial dysplasia transformation, and extends survival. This work establishes a previously unrecognized SOX2-CCL2-IL1 pathway that leads to irreversible immune escape when precancerous epithelial lesions transform.

cancer biology↗

bioRxiv2024

Naturally ornate RNA-only complexes revealed by cryo-EM

Myriad families of natural RNAs have been proposed, but not yet experimentally shown, to form biologically important structures. Here we report three-dimensional structures of three large ornate bacterial RNAs using cryogenic electron microscopy at resolutions of 2.9-3.1 [A]. Without precedent among previously characterized natural RNA molecules, Giant, Ornate, Lake- and Lactobacillales-Derived (GOLLD), Rumen-Originating, Ornate, Large (ROOL), and Ornate Large Extremophilic (OLE) RNAs form homo-oligomeric complexes whose stoichiometries are retained at concentrations lower than expected in the cell. OLE RNA forms a dimeric complex with long co-axial pipes spanning two monomers. Both GOLLD and ROOL form distinct RNA-only multimeric nanocages with diameters larger than the ribosome. Extensive intra- and intermolecular A-minor interactions, kissing loops, an unusual A-A helix, and other interactions stabilize the three complexes. Sequence covariation analysis of these large RNAs reveals evolutionary conservation of intermolecular interactions, supporting the biological importance of large, ornate RNA quaternary structures that can assemble without any involvement of proteins.

biophysics↗

bioRxiv2024

Isolation and Characterization of Antibodies Against VCAM-1 Reveals Putative Role for Ig-like Domains 2 and 3 in Cell-to-Cell Interaction

Vascular cell adhesion molecule-1 (VCAM-1) plays an important role in inflammation, where it facilitates the recruitment of leukocytes to the inflamed area via leukocytes VLA-4 and endothelial cells VCAM-1 interaction. VCAM-1 expression is also upregulated in certain cancers. VCAM-1 has 7 Ig-like domains, with domains 1 and 4 shown to be critical for VLA-4 binding. However, the specific functions of individual VCAM-1 Ig-like domains remain poorly understood. In this study, we identified single-chain variable fragment (scFvs) antibodies targeting domains 2, 3, and 5 of VCAM-1, and investigated the ability of these antibodies to block VCAM-1-mediated cell adhesion to macrophages. We show that scFv antibodies against Ig-like domains 2 and 3 significantly interfere with the ability of macrophages to bind endothelial cells, suggesting that these domains also play a role in facilitating this interaction. These results emphasize the need to more carefully study the role of each domain on VCAM-1 function and highlight the potential of targeting these VCAM-1 domains for more tailored therapeutic interventions in inflammatory diseases and cancer.

molecular biology↗

bioRxiv2024

One score to rule them all: regularized ensemble polygenic risk prediction with GWAS summary statistics

Ensemble learning has become a cornerstone for improving the predictive accuracy of polygenic risk scores (PRS), and nearly all recent multi-ancestry PRS methods incorporate ensemble learning as a final step. However, existing ensemble approaches require individual-level genotype data for model training, which limits their real-world applications, especially in non-European populations without sufficient genomic samples. Here, we introduce a statistical framework for constructing regularized ensemble PRS that integrates a large number of candidate PRS models using only genome-wide association study summary statistics. Through extensive analyses across multiple traits and populations, we demonstrate that our method consistently outperforms state-of-the-art PRS approaches within and across ancestries. This framework presents "one score to rule them all" for its capability to enable seamless integration of newly developed PRS models with existing ones, providing a scalable and general solution for future PRS development and application.

genetics↗

bioRxiv2024

Rhizobium rhizogenes A4-derived strains mediate hyper-efficient transient gene expression in Nicotiana benthamiana and other solanaceous plants

Agroinfiltration, a method utilizing agrobacteria to transfer DNA into plant cells, is widely used for transient gene expression in plants. Besides the commonly used Agrobacterium strains, Rhizobium rhizogenes can also introduce foreign DNA into host plants for gene expression. While many R. rhizogenes strains have been known for inducing hairy root symptoms, their use for transient expression has not been fully explored. Here, we showed that R. rhizogenes A4 outperformed all other tested agrobacterial strains in agroinfiltration experiments on leaves of Nicotiana benthamiana and other solanaceous plants. By conducting an agroinfiltration screening in N. benthamiana leaves using various agrobacterial strains carrying the RUBY reporter gene cassette, we discovered that A4 mediates the strongest and fastest transient expression. Utilizing the genomic information, we developed a collection of disarmed and modified strains derived from A4. By performing vacuum infiltration assays, we demonstrated that these A4-derived strains efficiently transiently transform 6-week-old N. benthamiana leaves, showing less sensitivity to the age of plants compared to the laboratory strain GV3101. Furthermore, we performed agroinfiltration using AS109, an A4-derived disarmed strain, on the leaves of tomato, pepper, and eggplant. Remarkably, AS109 mediated transient gene expression on tested solanaceous plants more effectively than all the tested commonly used agrobacterial strains. This discovery paves the way for establishing R. rhizogenes A4-derived strains as a new option for enhancing transient expression in N. benthamiana and facilitating the functional study of plant genes in other solanaceous species.

plant biology↗