Search bioRxiv⌕ Search

Biology subjects

Pan, Q.

Publications and source records attributed to Pan, Q..

At least 19 recordsLinked to original sources

CharacTERT: A machine learning tool for classifying hTERT missense variants

Missense mutations in TERT, the gene encoding the human telomerase catalytic subunit hTERT, are associated with Telomere Biology Disorders (TBDs). Experimentally elucidating the effects of all possible missense variants would be time-consuming and technically challenging. Moreover, current computational predictors are not hTERT-specific and primarily rely on sequence information, failing to capture the complex biological and structural context of the telomerase enzyme. In this work, we developed three machine learning models integrating both sequence- and structure-based features to account for the biological mechanisms of hTERT. Compared to state-of-the-art methods, our best-performing models achieved a higher Matthews Correlation Coefficient of 0.88 on ClinVar and gnomAD curated variants and demonstrated robust sensitivity (0.75) on a dataset curated according to guidelines from the American College of Medical Genetics and Genomics and Association for Molecular Pathology (ACMG/AMP). Feature interpretation highlighted hTERT residue conservation and changes in hydrophobic and weak polar interactions as critical determinants of pathogenicity. Finally, in silico saturation mutagenesis was performed to present a mutational landscape of TERT, available in a user-friendly web server, CharacTERT, which could offer valuable insights into the molecular mechanisms driving TBDs, aid in early diagnosis, as well as guide personalized treatment strategies. CharacTERT is freely available at https://biosig.lab.uq.edu.au/charactert/.

bioinformatics↗

Female-biased sex ratios despite stable genetic sex determination across a climatic gradient in a marine fish

Adult sex ratio (ASR) is a fundamental parameter shaping population dynamics and evolutionary trajectories. In theory, ASR is governed by the sex determination (SD) system and sex-specific mortality. Yet, ASR is often hard to predict in natural populations because of ecological effects on its determinants. We address this ecological complexity by investigating the drivers of ASR across a steep climatic gradient in Scandinavian populations of the two-spotted goby Pomatoschistus flavescens. Demographic surveys accounting for over 25000 fish in 30 locations, spanning 10{degrees} of latitude, revealed a general female-bias of the ASR, ca. 75%. To assess the contribution of SD, we combined reduced representation sequencing of 180 adults with our novel long-read male genome assembly. We characterised the genetic architecture of an XX/XY SD system, and identified a male-specific duplicate from the TGF-{beta} signaling pathway, amhr2y, as a candidate sex-determining gene. Subsequent genotyping of eggs and juveniles showed consistently unbiased sex ratios across latitudes, implying that the female-biased arises at later life stages. Interpreted in the light of historical data, our findings allow us to reject environmental sex determination and sex-biased mortality as important drivers of ASR bias in this species. We propose instead that the female bias originates from sex-specific patterns of habitat distribution, which could result from ecological conditions interacting with the distinct reproductive strategies of females and males. Our results underscore the need to go beyond primary theoretical expectations to understand fundamental demographic parameters within their relevant ecological context.

evolutionary biology↗

LDLR variant classification through activity-normalized prime editing screening

BackgroundInherited variants in the LDL receptor (LDLR) gene are the most common cause of familial hypercholesterolemia (FH), significantly increasing coronary artery disease risk. Early identification of pathogenic LDLR variants enables prompt intervention with lipid-lowering therapies; however, the majority of LDLR variants observed in the population have uncertain or absent clinical classifications, limiting the potential to improve clinical management. MethodsWe developed an innovative, activity-normalized prime editing screening pipeline to measure the impact of 5,184 LDLR coding variants on LDL-cholesterol (LDL-C) uptake. Through pairing a genotypic outcome reporter with every prime editing guide RNA (pegRNA), we adjust phenotypic measurements to account for variable editing efficiency, extending activity normalization to prime editing for the first time at this scale. Further, we use a statistical estimation approach that leverages measurements for all missense variants at a given position to denoise the resulting scores. ResultsWe show that prime editing-mediated reporter editing correlates with endogenous variant installation frequency, allowing activity normalization to improve imputation of LDLR variant effect. Our optimized prime editing assay identifies a broad, continuous spectrum of variant functional effects. We achieve robust separation of pathogenic vs. benign ClinVar variants and concordance between experimentally derived functional scores and LDL-C levels measured in UK Biobank participants. Further, when calibrating the strength of evidence provided by this functional screening data to align with the ACMG/AMP variant interpretation guidelines, and integrating additional sources of evidence, a majority of currently unclassified rare LDLR variants meet evidence thresholds for reclassification. We use the broad coverage of this screen to gain insight into how apolipoproteins bind to LDLR. In particular, we identify and characterize rare LDLR variants that enhance LDL-C uptake through increased interaction with apolipoprotein B. Finally, we compare prime editing-based functional scores with those derived from recent base editing and cDNA-based LDLR variant screens, showing that these approaches all show robust correlation with clinically observed LDL-C levels and computational scores, while prime editing identifies candidate splice-altering coding variants that are not modeled by cDNA screening. ConclusionsAltogether, our approach demonstrates the power of prime editing to significantly improve understanding of how variants in LDLR impact function and contribute to FH.

genetics↗

The effects of denifanstat on hepatitis E virus replication and triggered inflammatory response

Metabolic dysfunction-associated steatotic liver disease (MASLD) is initiated by intra-cellular fatty acid accumulation, with macrophage-mediated inflammatory responses playing a pivotal role in driving disease progression. MASLD frequently coexists with viral infections, for example in the context of hepatitis E virus (HEV) infection which explores host lipid metabolism for its replication and secretion. But no targeted therapies currently exist for this complex comorbidity. Denifanstat, a fatty acid synthase (FASN) inhibitor developed for treating metabolic dysfunction-associated steatohepatitis (MASH), has shown to have antiviral activity against SARS-CoV-2 infection. In this study, we aim to assess the effects of denifanstat on HEV infection in the context of steatotic liver disease. We first employed human liver-derived primary organoids exposed to fatty acids and inoculated with HEV. Treatment with denifanstat had no major effect on HEV replication in the steatotic organoids. However, we found that denifanstat moderately inhibited viral replication in macrophages, and regulated HEV-triggered inflammatory responses. These results highlight the importance of understanding how the emerging MASH treatment may affect viral infections that coexist in patients.

microbiology↗

Enterococcus faecalis delivers Obg GTPase via extracellular vesicles to instigate mTOR activity and promote HCC tumorigenesis

Hepatocellular carcinoma (HCC) is a malignant tumor that has been associated with dysbiosis of the gut microbiota. However, how the gut microbiota plays an oncogenic role in HCC remains largely unknown. Here, we show that Enterococcus faecalis (E. faecalis) is highly enriched in liver tumor tissues and is positively correlated with pathogenesis of HCC. E. faecalis promotes liver cancer cell proliferation, protein translation, cell migration and tumorigenesis. Mechanistically, we found that EF-derived extracellular vesicles (EF-EVs) deliver EF-Obg GTPase to activate host mTOR, thereby promoting liver cancer progression. Intriguingly, the EF-Obg protein exerts its influence on the mTOR pathway via a Ras-like G domain involved in GTP binding. EF-Obg exhibits a striking homology with the G1 site of the G domain of Rheb, a key positive regulator of mTOR. Obg gene is critical for E. faecalis to activate mTOR to promote hepatocarcinogenesis based on an engineered obg knockdown strain by CRISPR interference experiment. Clinically, abundant EF-Obg protein expression is correlated with enhanced activation of mTOR, leading to poor overall survival in HCC patients. Significantly, treatment of mTOR inhibitor Everolimus confers effectiveness in EF-colonized liver cancer orthotopic model, suggesting that Everolimus therapeutic approach can be effective for liver cancer patients with enriched E. faecalis. Taken together, we provide mechanistic and functional evidence to verify a direct causal relationship between tumor-resident E. faecalis enrichment and liver carcinogenesis, revealing that EF-Obg functions as a previously unidentified cross-kingdom activator of mTOR to promote liver tumorigenesis.

cell biology↗

Mapping the impact of microplastics exposure on enteric viral infections in intestinal organoid models

Micro- and nanoplastics (MNPs) are pervasive environmental pollutants increasingly detected in human tissues, yet their long-term health consequences remain poorly understood. The intestinal epithelium, continuously exposed to ingested MNPs and serving as a primary entry site for enteric viruses, provides a critical context to study pollutant-pathogen interactions. We used human intestinal organoids to model chronic exposure, found that prolonged NPs exposure potentially triggered mitochondrial stress and broad metabolic disruption without overt toxicity. This reprogramming altered the host-virus response and also reduced sensitivity to the antiviral treatment. These findings suggest that chronic plastic exposure may subtly but persistently reshape mucosal physiology in ways that alter host-pathogen response and therapeutic efficacy, highlighting an urgent need to address environmental pollutants in infectious disease and public health research.

cell biology↗

CircRNA-based, non-integrated, in vivo panCAR-mediated B cell immune resetting in mouse models and non-human primates

B cell immune resetting, defined as the comprehensive depletion and subsequent reconstitution of B cells, represents a pivotal therapeutic approach for B cell-associated disorders. Recently, chimeric antigen receptor T cell (CAR-T) therapy has demonstrated substantial clinical efficacy in treating B cell malignancies and autoimmune diseases. However, this approach remains constrained by its reliance on lymphodepleting conditioning regimens, high invasiveness, high costs, and the risk of severe cytokine release syndrome (CRS). Antibody-based therapies, including monoclonal antibodies and T cell-engager (TCE) bispecific antibodies, suffer from suboptimal tissue penetration, resulting in inadequate B cell depletion in lymphoid and peripheral tissues, and subsequent rapid resurgence of pathogenic B cells. Here, we engineered highly stable circular RNAs (circRNAs) encoding anti-CD19 CAR (circRNA-CAR), which demonstrated prolonged and enhanced CAR protein expression compared to conventional linear mRNAs. By employing immune cell-tropic lipid nanoparticles (LNPs) for in vivo delivery, circRNA-CAR facilitated the generation of panCAR immune cells (including CAR-T, CAR-NK, and CAR-macrophages) in both murine models and cynomolgus monkeys. Compared to rituximab or teclistamab, in vivo panCAR induced more extensive B cell depletion across multiple tissue compartments in mice. Moreover, in vivo panCAR mediated robust B cell depletion, eliminated autoreactive antibodies, and restored renal function by alleviating proteinuria in a systemic lupus erythematosus (SLE) murine model. It also effectively reduced IgE antibody levels, decreased eosinophil activity, and improved lung pathology in an asthma murine model. Notably, in vivo panCAR-mediated B cell depletion significantly attenuated aging-associated phenotypes in 17- month-old aging mice, remarkably outperforming both rituximab and teclistamab. In cynomolgus monkeys, in vivo panCAR achieved sustained B cell depletion lasting approximately 40 to 50 days, followed by reconstitution of a naive B cell repertoire. Collectively, circRNA-based in vivo panCAR enables complete B cell immune resetting, offering a versatile and promising therapeutic platform for a broad spectrum of B cell- and antibody-mediated disorders.

immunology↗

An improved SIFT algorithm based method for detecting of tumor tissue movement in respiratory motion

BackgroundThis paper intends to make use of the excellent performance of the SIFT algorithm to accurately calculate the displacement of tumor tissues during respiratory movement. Material and methodFirst, we moved phantoms to up,down,right,left respectively to simulate the different directions of movement during respiratory. And rotated phantom to left and right respectively by the angle of 5,10,30,45. Then, used SIFT and cross-correlation algorithms to search the phantoms. Last, use the t-test to check the differences between different directions. ResulThe t-test result proved that there was no significant difference in the detection of moving phantom in all directions. There was no statistical difference between the SIFT of the moving motif and the cross-correlation. The detection results of rotating phantom indicated that the SIFT test results could be used to detect distortion. ConclusionSIFT algorithm can be used to calculate the distortion caused by the thoracic tissue during respiratory movement.

biophysics↗

Cell Type Specific Aging Transcriptional Signatures of Human Retina Through Integrated Machine Learning and Single-Cell Transcriptomics

PurposeTo characterize cell type specific transcriptional changes during human retinal aging and develop machine learning model for cellular age discrimination in a Chinese cohort. DesignCross-sectional, laboratory-based observational study. ParticipantsEighteen unfrozen retinas from 12 Chinese donors (9 young, 34-55y; 9 old, 68-92 y). MethodsSingle-cell RNA sequencing (10x, v3.1) generated 223612 cells, batch-corrected with scVI; age-related signatures were defined by intersecting single-cell and pseudo-bulk differentially expressed genes, then cell-type-specific panels were rank-ordered with L1-regularised logistic regression plus recursive feature elimination and interpreted through hallmark-pathway enrichment and transcription-factor regulon mapping. Main Outcome MeasuresAge-related cellular composition shifts; cell-type-specific differentially expressed genes; machine-learning classifier accuracy and feature rankings; transcription factor regulon activity changes. ResultsEleven major retinal cell populations were identified. Aging showed declining rod-to-cone ratios, reduced bipolar cell proportions among interneurons, and increased astrocyte abundance. Muller glial cells exhibited the most pronounced transcriptional changes, followed by bipolar cells and rods. Machine-learning classifiers achieved 80-96% accuracy across cell types (microglia 96%, horizontal cells 93%, bipolar cells 91%, cones 90%, rods 89%). Shared aging signatures included mitochondrial dysfunction and inflammatory activation. Cell specific vulnerabilities emerged: mitochondria-centric stress in rods/bipolar cells, proteostasis-retinoid metabolism in cones, and structural-RNA maintenance in horizontal cells. ConclusionsThis study provides the first machine learning derived, cell-type specific aging signatures for human retina in a Chinese cohort, revealing both conserved molecular hallmarks and distinctive cellular vulnerabilities that inform targeted therapeutic strategies for retinal aging.

bioinformatics↗

Convergent Evolution of H4K16ac-mediated Dosage Compensation Shapes Sex-dependent Lifespan in a ZW Species

Sex chromosomes impact chromatin organization and histone modification dynamics differently between males and females, particularly those involved in dosage compensation (DC). The stability of DC mechanisms may shape sex-specific phenotypes and traits such as lifespan. However, the tissue- and age-dependent variations as well as the evolutionary diversity of DC mechanisms are incompletely understood. Here, we investigate the occurrence of histone H4 lysine 16 acetylation (H4K16ac), previously known for its role in sex chromosome DC in the male-heterogametic fruit fly Drosophila melanogaster and the green anole lizard Anolis carolinensis. By sampling multiple arthropods, we find the convergent evolution of H4K16ac for DC in a female-heterogametic (ZW) species, the crustacean Artemia franciscana. CUT&Tag analysis demonstrates that H4K16ac is confined to the non-recombining stratum of the Z chromosome in females. H4K16ac-mediated DC is established during embryogenesis, sustained until adulthood, but then becomes reconfigured and variable in aged females. Interfering with acetylation alleviates male-female differences in lifespan. Our findings shed light on the evolutionary diversity of DC across species. They highlight histone acetylation as a sex-specific vulnerability linked to the chromatin landscape of sex chromosomes and its potential role in driving male-female differences.

evolutionary biology↗

Single-cell transcriptomic atlas of human retina from Chinese donors reveals population-specific cellular diversity

The human retina exhibits complex cellular heterogeneity which is critical for visual function, yet comprehensive ethnic-specific references are scarce in ophthalmic transcriptomics. The lack of single-cell RNA sequencing (scRNA-seq) data from Asian populations particularly Chinese donors imposes significant limitations in understanding population-specific retinal biology. We constructed the first comprehensive single-cell transcriptomic atlas of the human retina from Chinese donors, generated through high-throughput scRNA-seq of [~]290,000 viable cells obtained from 18 fresh retinal specimens (living donor and post-mortem specimens). Our multi-level analysis identified 10 distinct retinal cell types, encompassing all major neuronal lineages, Muller glia, astrocytes, microglia. Detailed subcluster analyses further revealed extensive heterogeneity, identifying distinct subtypes within several cell populations such as 7 amacrine cell subtypes and 14 bipolar cell subtypes. Concurrently, through systematic analysis of delineated subtype-specific molecular programs, we mapped their associated biological signaling pathways, functions, and mechanistic processes. This analysis explained the critical involvement of these subpopulations in core biological processes including synaptic organization, neurotransmission, and phototransduction cascades, potentially governing retinal homeostatic regulation and disease mechanisms. Single-cell transcriptomic atlas of the human retina from Chinese donors describes a comprehensive cellular landscape, encompassing major cell types and subtypes including neuronal, glial, and immune populations. This ethnic-specific atlas provided an important reference for understanding retinal development, cellular interaction and disease pathogenesis in Chinese populations, addressing a longstanding gap in ophthalmic transcriptomic resources.

genomics↗

Single-Cell Transcriptomics Reveals Dynamic Microglial States and Neural-Immune Networks in Human Diabetic Retinopathy

Diabetic retinopathy (DR) is a major cause of vision loss worldwide. Here, we conduct single-cell RNA sequencing of twenty human retina samples (from living and post-mortem donors) across non-diabetic, diabetic, and DR states to create a comprehensive transcriptomic atlas. We identify two stable microglial populations--homeostatic and inflammatory--that exist along a functional continuum, plus a neutrophil cluster within C1QA+ myeloid cells with dynamic transitions occurring throughout disease progression. Module-level analysis reveals divergent transcriptional trajectories: homeostatic microglia maintain energetic programs while selectively upregulating stress elements, whereas inflammatory microglia layer additional pro-inflammatory programs onto preserved biosynthetic foundations. Eleven co-expression modules organize into two major axes: an inflammatory-stress axis, and a regulatory/metabolic-motility axis, with a stable translation module persisting across disease stages. Cell communication analysis further highlights sophisticated neural-immune interactions, particularly between photoreceptors and microglia. Our findings provide insights into the complex cellular dynamics of DR progression and suggest potential therapeutic targets for early intervention.

bioinformatics↗

A novel SARS-CoV-2-derived infectious vector system

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of COVID-19. The development of antiviral drugs for COVID-19 has been hampered by the requirement of a biosafety level 3 (BSL3) laboratory for experiments related to SARS-CoV-2, and by the lack of easy and precise methods for quantification of infection. Here, we developed a SARS-CoV-2 viral vector composed of all four SARS-CoV-2 structural proteins constitutively expressed in lentivirally transduced cells, combined with an RNA replicon deleted for SARS-CoV-2 structural protein genes S, M and E, and expressing a luciferase-GFP fusion protein. We show that, after concentrating viral stocks by ultracentrifugation, the SARS-CoV-2 viral vector is able to infect two human cell lines expressing receptors ACE2 and TMPRSS2. Both luciferase activity and GFP fluorescence were detected, and transduction was remdesivir-sensitive. We also show that this vector is inhibited by three type I interferons (IFN-I) subtypes. Although improvements are needed to increase infectious titers, this vector system may prove useful for antiviral drug screening and SARS-CoV-2-related investigations.

microbiology↗

Fresh Human Retinal scRNA-seq Atlas Reveals a Novel Cone Subtype and Cellular Diversity

BackgroundCurrent human retina studies predominantly utilize post-mortem tissue, and the sample accessibility constraints make the characterization of the living human retina at single-cell resolution a challenge. Although single-nucleus RNA-seq expands the utility of frozen samples, it provides a nuclear-centric view, potentially missing key cytoplasmic information and transient biological processes. Thus, it is important to generate resources directly from living human retinal tissue to complement existing datasets. MethodsWe profiled 106,829 single cells from nine unfrozen human retina samples. Living samples were collected withinLJ10LJmin of therapeutic enucleation and four postmortem samples were collected within 6LJh. After standardized dissociation, single-cell transcriptomes were generated using 10x Genomics 3 RNA-seq and applied scVI to generate batch-corrected integrated atlas. Major cell types and subtypes were annotated through iterative Leiden clustering, canonical markers. Subsequent analyses included differential expression comparisons between cell states and regulon activity profiling to further characterize cellular identities and regulatory networks. Transcriptional dynamics were assessed using RNA velocity, and cell-cell signaling pathways were inferred with CellChat. Key findings were validated in independent samples from two additional donors (four samples) using the identical workflow. ResultsWe contribute to establishing a reference for retinal cell type proportions and cellular states. Our analysis revealed ELF1-mlCone, a distinct cluster of mlCone photoreceptors identified by distinct transcriptional features. The presence and transcriptional features of this cluster were validated in independent samples. Additionally, by comparing living and post-mortem samples, our study highlights differences in transcriptional dynamics: living tissue preserved coherent RNA velocity streams, enabling clear dynamic state transitions, while post-mortem tissue exhibited disorganized patterns. These findings suggest that using living tissue can improve the capture of active cellular states and transitions. ConclusionsOur atlas provides a single-cell reference contrasting living versus early post-mortem human retina, integrating cell type composition, transcriptional diversity, and functional insights. It may serve as a useful resource for retinal research and for understanding aspects of human retinal biology, particularly given its inclusion of living tissue and diverse pathological states.

bioinformatics↗

DeepSME: De Novo Nanopore Basecalling of Motif-insensitive DNA Methylation and Alignment-free Digital Information Decryptions at Single-Molecule Level

Hidden messages in DNA molecules by employing chemical modifications has been suggested for private data storage and transmission at high information density. However, rapidly decoding these "molecular keys" with corresponding basecallers remains challenging. We present DeepSME, a nanopore sequencing and deep-learning based framework towards single-molecule encryption, demonstrated by using 5-hydroxymethylcytosine (5hmC) substitution for individual nucleotide recognition rather than sequential interactions. This non-natural, motif-insensitive methylation disrupts ion current, resulting in a readout failure of 67.2%-100%, concealing the privacy within the DNAs. We further develop an alignment-free DeepSME basecaller as a key to reconstitute the digital information. Our three-stage training pipeline, expands k-mer size from 46 to 49, achieving over 92% precision and recall from scratch. DeepSME deciphers fully 5hmC concealed text and image within 16x coverage depth with an F1-score of 86.4%, surpassing all the state-of-the-art basecallers. Demonstrated on edge computing devices, DeepSME holds supreme potential for DNA-based private communications and broader bioengineering and medical applications.

bioinformatics↗

Composite Hedges Nanopores: A High INDEL-Correcting Codec System for Rapid and Portable DNA Data Readout

DNA, as the origin for the genetic information flow, has also been a compelling alternative to non-volatile information storage medium. Reading digital information from this highly dense but lightweighted medium nowadays relied on conventional next-generation sequencing (NGS), which involves wash and read cycles for synchronization and the indel (insertion and deletion) errors rarely occur. However, these time-consuming cycles hinder the future of real-time data retrieval. Nanopore sequencing holds the promise to overcome the efficiency problem, but high indel error rates lead to the requirement of large amount of high-quality data for accurate readout using emerging NGS-based codec systems. Here we introduce Composite Hedges Nanopores (CHN), a nanopore-based codec scheme tailored for real-time data retrieval, capable of handling indel rates up to 15.9% and substitution rates up to 7.8%. The overall information density can be doubled from 0.59 to 1.17 by utilizing a degenerated eight-letter alphabet, where one composite strand will be projected into eight normal strands. We demonstrate that sequencing times of 20 and 120 minutes were sufficient for processing representative text and image files (7 and 115 composite strands), respectively. The time-diminishing deviations are mainly originated from the extremely uneven abundance among the composite strands (cross-group variation) as well as the huge inequality among the normal strands (in-group variation). Moreover, to achieve complete data recovery, it is estimated that text and image data require 4x and 8x physical redundancy (coverage) of composite strands, respectively. Our CHN codec system excels on both molecular design and equalized dictionary usage, laying a solid foundation for nucleic acid-based data retrieval and encoding approaching to real-time, applicable in both cloud and edge computing systems.

bioinformatics↗

Epigenetic delineation of the earliest cardiac lineage segregation by single-cell multi-omics

The mammalian heart is formed from multiple mesoderm-derived cell lineages. However, it remains largely unknown when and how the specification of mesoderm towards cardiac lineages is determined. Here, we systematically depict the transcriptional trajectories toward cardiomyocyte in early mouse embryo, and characterize the epigenetic landscapes underlying the early mesodermal lineage specification by single-cell multi-omics analyses. The analyses also reveal distinct core regulatory networks (CRN) in controlling specification of mesodermal lineages. We further demonstrate the essential role HAND1 and FOXF1 in driving the earliest cardiac progenitors specification. These key transcription factors occupy at distinct enhancers, but function synergistically and hierarchically to regulate the expression of cardiac-specific genes. In addition, HAND1 is required for exiting from the nascent mesoderm program, while FOXF1 is essential for driving cardiac differentiation during juxta-cardiac field (JCF) specification. Our findings establish transcriptional and epigenetic determinants specifying the early cardiac lineage, providing insights for the investigation of congenital heart defects.

molecular biology↗

Non-additive dosage-dependent effects of TaGS3 gene editing on grain size and weight in wheat

The grain size in cereals is one of the main component traits contributing to yield. Previous studies showed that loss-of-function (LOF) mutations in GS3, encoding G{gamma} subunit of the multimeric G protein complex, increase grain size and weight in rice. While association between allelic variation in GS3 homologs of wheat and grain weight/size was detected previusly, the effects of LOF alleles on these traits remain unknown. We used genome editing to create the TaGS3 mutant lines with the LOF homeo-allele dosage variation. Contrary to results obtained for rice, editing of all three TaGS3 copies result in significant decrease in grain length, width, grain area and weight, without affecting number of grains per spike. Compared to wild type, the highest increase in grain weight and area was observed in mutants with the intermediate dosage of the LOF alleles, indicating that suppressive effects of TaGS3 on grain size and weight in wheat are dosage-dependent and non-additive. Our results suggest that TaGS3 likely represents a functionally diverged homolog of GS3 evolved in the wheat lineage. The newly developed LOF alleles of TaGS3 expand the set of CRISPR-Cas9-induced variants of yield component genes that could be used for increasing grain weight in wheat.

genetics↗