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3.
Aging (Albany NY) ; 162024 Sep 16.
Article in English | MEDLINE | ID: mdl-39287993

ABSTRACT

Alzheimer's Syndrome (AD) is a neurodegenerative disease that is prevalent in middle-aged and elderly people. As the disease progresses, patients gradually lose the ability to take care of themselves, which brings a heavy burden to the family. There is a link between leukocyte telomere length (LTL) and cognitive ability. To search for possible pathogenic mechanisms and potential therapeutic agents, we demonstrated a causal link between LTL and AD using Mendelian randomization analysis (MR). The expression of the target gene NBR2 and the downstream mRNA GJA1 and GJA1-related genes, pathway enrichment, and association with immune cells were further explored. Using the gene cluster-drug target interaction network, we obtained potential therapeutic drugs. Our study provides evidence for a causal link between AD and LTL, suggesting medicines that may treat and alleviate AD symptoms.

4.
J Hazard Mater ; 480: 135968, 2024 Sep 26.
Article in English | MEDLINE | ID: mdl-39342845

ABSTRACT

Sugar beet (Beta vulgaris L.) shows potential as an energy crop for cadmium (Cd) phytoremediation. To elucidate its in vivo response strategy to Cd exposure, seedlings were treated with 1, 3, and 5 mmol/L CdCl2 (Cd-1, Cd-3, and Cd-5) for 6 h, using 0 mmol/L CdCl2 (Cd-0) as the control. The results showed that Cd-3 promoted a unique "hormesis" effect, leading to superior growth performance, increased levels of chlorophyll, soluble protein, and SOD activity, and reduced MDA content in sugar beet, compared to Cd-1, Cd-5, and even Cd-0. GO and KEGG enrichments and PPI networks of transcriptomic analysis revealed that the differentially expressed genes (DEGs) were primarily involved in lipid metabolism, cellular protein catabolism, and photosynthesis. Notably, the MAPK signaling pathway was significantly enriched only under Cd-3, with the up-regulation of ABA-related core gene BvPYL9 and an increase in ABA content after 6 h of Cd exposure. Furthermore, overexpression of BvPYL9 in Arabidopsis thaliana (OE-1 and OE-2) resulted in enhanced growth (fresh weight, dry weight, and root length), as well as higher ABA and soluble protein contents under different Cd treatments. Cd-induced transcriptional responses of BvPYL9 were also evident in OE-1 and OE-2, especially at 10 µmol/L, indicated by qRT-PCR. These findings suggest that ABA-mediated MAPK signaling pathway is activated in response to Cd toxicity, with BvPYL9 being a key factor in the cascade effects for the Cd-induced hormesis in sugar beet.

5.
Org Lett ; 2024 Sep 27.
Article in English | MEDLINE | ID: mdl-39331679

ABSTRACT

Asymmetric aza-Heck cyclization and coupling reactions offer efficient access to enantioenriched N-heterocycles, yet the current studies focus primarily on sequential C-N and C-C bond formation. Herein, we report an enantioselective reductive aza-Heck cyclization followed by a C-S coupling sequence, ultimately yielding sulfide-containing enantioenriched pyrrolines. The reaction is conducted under mild conditions and tolerates broad functionalities including alkynes, phenols, anilines, amides, nitriles, and bromides.

6.
Insights Imaging ; 15(1): 226, 2024 Sep 19.
Article in English | MEDLINE | ID: mdl-39320574

ABSTRACT

OBJECTIVE: To establish a practical risk stratification system (RSS) based on ultrasonography (US) and clinical characteristics for predicting soft tissue masses (STMs) malignancy. METHODS: This retrospective multicenter study included patients with STMs who underwent US and pathological examinations between April 2018 and April 2023. Chi-square tests and multivariable logistic regression analyses were performed to assess the association of US and clinical characteristics with the malignancy of STMs in the training set. The RSS was constructed based on the scores of risk factors and validated externally. RESULTS: The training and validation sets included 1027 STMs (mean age, 50.90 ± 16.64, 442 benign and 585 malignant) and 120 STMs (mean age, 51.93 ± 17.90, 69 benign and 51 malignant), respectively. The RSS was constructed based on three clinical characteristics (age, duration, and history of malignancy) and six US characteristics (size, shape, margin, echogenicity, bone invasion, and vascularity). STMs were assigned to six categories in the RSS, including no abnormal findings, benign, probably benign (fitted probabilities [FP] for malignancy: 0.001-0.008), low suspicion (FP: 0.008-0.365), moderate suspicion (FP: 0.189-0.911), and high suspicion (FP: 0.798-0.999) for malignancy. The RSS displayed good diagnostic performance in the training and validation sets with area under the receiver operating characteristic curve (AUC) values of 0.883 and 0.849, respectively. CONCLUSION: The practical RSS based on US and clinical characteristics could be useful for predicting STM malignancy, thereby providing the benefit of timely treatment strategy management to STM patients. CRITICAL RELEVANCE STATEMENT: With the help of the RSS, better communication between radiologists and clinicians can be realized, thus facilitating tumor management. KEY POINTS: There is no recognized grading system for STM management. A stratification system based on US and clinical features was built. The system realized great communication between radiologists and clinicians in tumor management.

7.
Clin Cosmet Investig Dermatol ; 17: 1999-2007, 2024.
Article in English | MEDLINE | ID: mdl-39247134

ABSTRACT

Rosacea is a common dermatosis with multiple pathogeneses, among which, rosacea fulminans may serve as a rare but severe subtype. This inflammatory disease usually presents as abrupt multiple erythema, pustules, and nodules localized on the face. Pregnancy and related changes of hormone levels may play a key role in the development and progression of the disease, although the exact mechanisms are unknown. In particular, treatment options, which includes systemic glucocorticosteroids, isotretinoin, and partial oral antibiotics, may be limited in pregnancy. Owing to the limited number of reported cases, standard diagnosis, treatment, and management guidelines remain unclear. Here, we report a case of rosacea fulminans happening in pregnancy treated successfully with oral erythromycin and short-term glucocorticosteroids, and share our review of the characteristics of RF cases during pregnancy.

8.
Angew Chem Int Ed Engl ; : e202412459, 2024 Sep 11.
Article in English | MEDLINE | ID: mdl-39261278

ABSTRACT

Inherently chiral calixarenes have garnered significant attention due to their distinctive properties, yet the development of efficient catalytic asymmetric synthesis methods remains a critical challenge. Herein, we report the asymmetric synthesis of calix[4]arenes featuring inherent or both inherent and axial chirality via a cobalt-catalyzed C-H activation/annulation strategy in high yield with excellent enantio- and diastereoselectivity (up to > 99% ee and > 20:1 dr). Electrooxidation was also suitable for this transformation to obviate the sacrificial metal oxidants, underscoring the environmentally friendly potential of this approach. A key octahedral cobaltacycle intermediate was synthesized and characterized, providing valuable insights into the mode of enantio- and diastereocontrol of this protocol. Noteworthy photoluminescence quantum yields of up to 0.94 were measured, underscoring the potential of these compounds in the domain of organic fluorescent materials.

9.
Front Nutr ; 11: 1401477, 2024.
Article in English | MEDLINE | ID: mdl-39267860

ABSTRACT

Background: The available data exhibit inconsistent findings regarding the association between obesity and sleep problems among older adults. The objective of this study was to assess the potential association between obesity and sleep disorders in the older population. Methods: The data utilised in this cross-sectional investigation was obtained from the National Health and Nutritional Examination Survey (NHANES) conducted between 2005 and 2018. The study employed a multivariate logistic regression model and conducted subgroup analysis to evaluate the association between obesity and sleep disturbance. Results: The investigation consisted of 2,570 older people who provided complete information, out of which 324 individuals satisfied the criteria for sleep disturbance. The findings from the multivariable adjusted logistic regression model indicate that individuals in the overweight and normal weight groups exhibited decreased odds of experiencing sleep disorder, as evidenced by the adjusted odds ratios (AOR) of 0.46 (95% confidence interval [CI] = 0.34-0.61) and 0.33 (95% CI = 0.22-0.47), respectively. These results were statistically significant (p < 0.001) when compared to individuals in the obese group. The investigation of age and gender subgroups demonstrated similar associations between various BMI categories and sleep disorders in the older population. Conclusion: In summary, there exists a correlation between obesity and sleep disorders in the senior population. A significant association was observed between BMI and the likelihood of experiencing sleep disorders, indicating a dose-response relationship. Individuals with a higher BMI demonstrated a heightened likelihood of experiencing sleep disorders compared to those with a lower BMI.

10.
Small ; : e2406206, 2024 Sep 13.
Article in English | MEDLINE | ID: mdl-39268781

ABSTRACT

Heart failure (HF) represents a cardiovascular disease that significantly threatens global well-being and quality of life. Electroactive nanomaterials, characterized by their distinctive physical and chemical properties, emerge as promising candidates for HF prevention and management. This review comprehensively examines electroactive nanomaterials and their applications in HF intervention. It presents the definition, classification, and intrinsic characteristics of conductive, piezoelectric, and triboelectric nanomaterials, emphasizing their mechanical robustness, electrical conductivity, and piezoelectric coefficients. The review elucidates their applications and mechanisms: 1) early detection and diagnosis, employing nanomaterial-based sensors for real-time cardiac health monitoring; 2) cardiac tissue repair and regeneration, providing mechanical, chemical, and electrical stimuli for tissue restoration; 3) localized administration of bioactive biomolecules, genes, or pharmacotherapeutic agents, using nanomaterials as advanced drug delivery systems; and 4) electrical stimulation therapies, leveraging their properties for innovative pacemaker and neurostimulation technologies. Challenges in clinical translation, such as biocompatibility, stability, and scalability, are discussed, along with future prospects and potential innovations, including multifunctional and stimuli-responsive nanomaterials for precise HF therapies. This review encapsulates current research and future directions concerning the use of electroactive nanomaterials in HF prevention and management, highlighting their potential to innovating in cardiovascular medicine.

11.
Pak J Med Sci ; 40(8): 1791-1796, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39281243

ABSTRACT

Objective: To compare the clinical effects of applying a 3D-printed ACT titanium trabecular intervertebral fusion cage and a Polyetheretherketone (PEEK) cage in posterior lumbar interbody fusion (PLIF). Methods: This was a clinical comparative study. Forty patients with degenerative lumbar diseases admitted at The Second People's Hospital of Dalian from January 2020 to December 2021 were selected and divided into an observation group (3D cage) and a control group (PEEK cage) using the random number table method, with each group of 20 cases. The visual analogue scale (VAS) scores, Japanese Orthopaedic Association (JOA) scores, Cobb angles at fusion segments, intervertebral height and intervertebral fusion situations of the patients between the groups were compared. Results: No significant differences were found in their operation time, intraoperative blood losses and operation related complications(p>0.05). In terms of postoperative VAS and JOA scores in both groups, they are all significantly improved compared with those before the operation, and their differences are also statistically significant(p<0.05). However, no statistical significance exists in inter-group differences(p>0.05). Postoperative Cobb angles and intervertebral height of patients in both groups are considerably bettered compared with those before the operation. Their differences show statistical significance(p<0.05), while inter-group differences are proved to be not statistically significant(p>0.05). Conclusions: Applying a 3D-printed ACT titanium trabecular intervertebral fusion cage or PEEK cage in PLIF has the potential to improve clinical symptoms of patients with degenerative lumbar diseases, and restore the Cobb angle and intervertebral height. 3D-printed ACT titanium trabecular intervertebral fusion cage can accelerate intervertebral fusion without increasing operation related complications.

12.
Nat Commun ; 15(1): 7135, 2024 Aug 20.
Article in English | MEDLINE | ID: mdl-39164290

ABSTRACT

The field of nickel catalysis has witnessed remarkable growth in recent years. However, the use of nickel catalysts in enantioselective C-H activation remains a daunting challenge because of their variable oxidation states, intricate coordination chemistry, and unpredictable reactivity patterns. Herein, we report an enantioselective C-H activation reaction catalyzed by commercially available and air-stable nickel(II) catalyst. Readily available and simple (S)-BINOL is used as a chiral ligand. This operationally simple protocol enables the synthesis of planar chiral metallocenes in high yields with excellent enantioselectivity through desymmetrization and kinetic resolution. Air-stable planar chiral nickelacycle intermediates are first synthesized via enantioselective C-H nickelation and shown to be possible intermediates of the reaction. Deuterium-labeling studies, alongside the characterization and transformation of chiral nickel(II) species, suggest that C-H cleavage is the enantio-determining step. Moreover, the large-scale synthesis and diverse synthetic transformations underscore the practicality of this protocol.

13.
World J Gastrointest Oncol ; 16(8): 3624-3634, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-39171164

ABSTRACT

BACKGROUND: Helicobacter pylori (H. pylori) infection can cause extensive apoptosis of gastric epithelial cells, serving as a critical catalyst in the progression from chronic gastritis, gastrointestinal metaplasia, and atypical gastric hyperplasia to gastric carcinoma. Prompt eradication of H. pylori is paramount for ameliorating the pathophysiological conditions associated with chronic inflammation of the gastric mucosa and the primary prevention of gastric cancer. Acacetin, which has multifaceted pharmacological activities such as anti-cancer, anti-inflammatory, and antioxidative properties, has been extensively investigated across various domains. Nevertheless, the impact and underlying mechanisms of action of acacetin on H. pylori-infected gastric mucosal epithelial cells remain unclear. AIM: To explore the defensive effects of acacetin on apoptosis in H. pylori-infected GES-1 cells and to investigate the underlying mechanisms. METHODS: GES-1 cells were treated with H. pylori and acacetin in vitro. Cell viability was assessed using the CCK-8 assay, cell mortality rate via lactate dehydrogenase assay, alterations in cell migration and healing capacities through the wound healing assay, rates of apoptosis via flow cytometry and TUNEL staining, and expression levels of apoptosis-associated proteins through western blot analysis. RESULTS: H. pylori infection led to decreased GES-1 cell viability, increased cell mortality, suppressed cell migration, increased rate of apoptosis, increased expressions of Bax and cle-caspase3, and decreased Bcl-2 expression. Conversely, acacetin treatment enhanced cell viability, mitigated apoptosis induced by H. pylori infection, and modulated the expression of apoptosis-regulatory proteins by upregulating Bcl-2 and downregulating Bax and cleaved caspase-3. CONCLUSION: Acacetin significantly improved GES-1 cell viability and inhibited apoptosis in H. pylori-infected GES-1 cells, thereby exerting a protective effect on gastric mucosal epithelial cells.

14.
Clin Respir J ; 18(8): e13814, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39129202

ABSTRACT

BACKGROUND: The primary cause of cancer-related fatalities globally is lung cancer. Although the chemotherapy drug cisplatin (DDP) has brought certain benefits to patients, the rapid development of drug resistance has greatly hindered treatment success. METHODS: We used the lung squamous cell carcinoma (LUSC) mRNA data set to explore the differentially expressed gene (RND1) in LUSC and detected RND1 expression in LUSC cells and DDP-resistant cells by qRT-PCR. Meanwhile, we performed abnormal expression treatment on RND1 and conducted CCK8, colony formation, and flow cytometry to evaluate the impact of RND1 expression on cell proliferation, apoptosis, and DDP resistance. In addition, we analyzed metabolism pathways involving RND1 using GSEA. We also used online tools such as hTFtarget and JASPAR to screen for the upstream transcription factor FOXA2 of RND1 and verified their relationship through CHIP and dual luciferase experiments. Finally, we validated the role of FOXA2-RND1 in DDP resistance in LUSC through the above experiments. RESULTS: RND1 was downregulated in LUSC, and overexpression of RND1 repressed proliferation and DDP resistance of LUSC cells and facilitated cell apoptosis. RND1 modulated the arachidonic acid (AA) metabolism pathway, and FOXA2 positively manipulated RND1 expression. By activating FOXA2, stabilizing RND1, and regulating AA levels, the sensitivity of LUSC cells to DDP could be enhanced. CONCLUSION: Our study suggested that FOXA2 positively modulated the RND1-AA pathway, which repressed the resistance of LUSC cells to DDP.


Subject(s)
Antineoplastic Agents , Arachidonic Acid , Carcinoma, Squamous Cell , Cell Proliferation , Cisplatin , Drug Resistance, Neoplasm , Gene Expression Regulation, Neoplastic , Lung Neoplasms , Humans , Cisplatin/pharmacology , Cisplatin/therapeutic use , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Drug Resistance, Neoplasm/genetics , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/pathology , Cell Proliferation/drug effects , Cell Line, Tumor , Arachidonic Acid/metabolism , Antineoplastic Agents/pharmacology , Apoptosis/drug effects
15.
Int J Mol Sci ; 25(16)2024 Aug 20.
Article in English | MEDLINE | ID: mdl-39201712

ABSTRACT

Nitrogen (N) is essential for sugar beet (Beta vulgaris L.), a highly N-demanding sugar crop. This study investigated the morphological, subcellular, and microRNA-regulated responses of sugar beet roots to low N (LN) stress (0.5 mmol/L N) to better understand the N perception, uptake, and utilization in this species. The results showed that LN led to decreased dry weight of roots, N accumulation, and N dry matter production efficiency, along with damage to cell walls and membranes and a reduction in organelle numbers (particularly mitochondria). Meanwhile, there was an increase in root length (7.2%) and branch numbers (29.2%) and a decrease in root surface area (6.14%) and root volume (6.23%) in sugar beet after 7 d of LN exposure compared to the control (5 mmol/L N). Transcriptomics analysis was confirmed by qRT-PCR for 6 randomly selected microRNAs, and we identified 22 differentially expressed microRNAs (DEMs) in beet root under LN treatment. They were primarily enriched in functions related to binding (1125), ion binding (641), intracellular (437) and intracellular parts (428), and organelles (350) and associated with starch and sucrose metabolism, tyrosine metabolism, pyrimidine metabolism, amino sugar and nucleotide sugar metabolism, and isoquinoline alkaloid biosynthesis, as indicated by the GO and KEGG analyses. Among them, the upregulated miR156a, with conserved sequences, was identified as a key DEM that potentially targets and regulates squamosa promoter-binding-like proteins (SPLs, 104889216 and 104897537) through the microRNA-mRNA network. Overexpression of miR156a (MIR) promoted root growth in transgenic Arabidopsis, increasing the length, surface area, and volume. In contrast, silencing miR156a (STTM) had the opposite effect. Notably, the fresh root weight decreased by 45.6% in STTM lines, while it increased by 27.4% in MIR lines, compared to the wild type (WT). It can be inferred that microRNAs, especially miR156, play crucial roles in sugar beet root's development and acclimation to LN conditions. They likely facilitate active responses to N deficiency through network regulation, enabling beet roots to take up nutrients from the environment and sustain their vital life processes.


Subject(s)
Beta vulgaris , Gene Expression Regulation, Plant , MicroRNAs , Nitrogen , Plant Roots , Beta vulgaris/genetics , Beta vulgaris/growth & development , Beta vulgaris/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Plant Roots/metabolism , Plant Roots/genetics , Plant Roots/growth & development , Nitrogen/metabolism , Nitrogen/deficiency , Acclimatization/genetics , Gene Expression Profiling
16.
ACS Appl Mater Interfaces ; 16(34): 45687-45694, 2024 Aug 28.
Article in English | MEDLINE | ID: mdl-39162076

ABSTRACT

Magnetoresistance effects are crucial for understanding the charge-spin transport as well as propelling the advancement of spintronic applications. Here, we report the coexistence of magnetic-moment-dependent (MD) and magnetic-field-driven (FD) unidirectional magnetoresistance (UMR) effects in CoFeB/InSb/CdTe heterostructures. The strong spin-orbital coupling of InSb and the matched impedance at the CoFeB/InSb interface warrant a distinct MD-UMR effect at room temperature, while the interaction between the in-plane magnetic field and the Rashba effect at the InSb/CdTe interface induces the marked FD-UMR signal that dominates the high-field region. Moreover, owning to different spin scattering mechanisms, these two types of non-reciprocal charge transports show opposite polarities with respect to the magnetic field direction, which further enables an effective phase modulation of the angular-dependent magnetoresistance. The demonstration of the tunable UMR response validates our CoFeB/InSb/CdTe system as a suitable integrated building block for multifunctional spintronic memory and sensor designs.

17.
Acta Biomater ; 186: 424-438, 2024 Sep 15.
Article in English | MEDLINE | ID: mdl-39122135

ABSTRACT

Mitophagy influences the progression and prognosis of ischemic stroke (IS). However, whether DNA methylation in the brain is associated with altered mitophagy in hypoxia-injured neurons remains unclear. Here, miR-138-5p was found to be highly expressed in exosomes secreted by astrocytes stimulated with oxygen and glucose deprivation/re-oxygenation (OGD/R), which could influence the recovery of OGD/R-injured neurons through autophagy. Mechanistically, miR-138-5p promotes the stable expression of Ras homolog enriched in brain like 1(Rhebl1) through DNA-methyltransferase-3a (DNMT3A), thereby enhancing ubiquitin-dependent mitophagy to maintain mitochondrial homeostasis. Furthermore, we employed glycosylation engineering and bioorthogonal click reactions to load mirna onto the surface of microglia and deliver them to injured region utilising the inflammatory chemotactic properties of microglia to achieve drug-targeted delivery to the central nervous system (CNS). Our findings demonstrate miR-138-5p improves mitochondrial function in neurons through the miR-138-5p/DNMT3A/Rhebl1 axis. Additionally, our engineered cell vector-targeted delivery system could be promising for treating IS. STATEMENT OF SIGNIFICANCE: In this study, we demonstrated that miR-138-5p in exosomes secreted by astrocytes under hypoxia plays a critical role in the treatment of hypoxia-injured neurons. And we find a new target of miR-138-5p, DNMT3A, which affects neuronal mitophagy and thus exerts a protective effect by regulating the methylation of Rbebl1. Furthermore, we have developed a carrier delivery system by combining miR-138-5p with the cell membrane of microglia and utilized the inflammatory chemotactic properties of microglia to deliver this system to the brain via intravenous injection. This groundbreaking study not only provides a novel therapeutic approach for ischemia-reperfusion treatment but also establishes a solid theoretical foundation for further research on targeted drug delivery for central nervous system diseases with promising clinical applications.


Subject(s)
DNA Methyltransferase 3A , MicroRNAs , Mitophagy , Neurons , Mitophagy/drug effects , MicroRNAs/genetics , MicroRNAs/metabolism , Neurons/metabolism , Neurons/drug effects , Animals , Cell Hypoxia/drug effects , Humans , DNA (Cytosine-5-)-Methyltransferases/metabolism , Exosomes/metabolism , Mice , Astrocytes/metabolism , Astrocytes/drug effects
18.
ACS Photonics ; 11(7): 2722-2728, 2024 Jul 17.
Article in English | MEDLINE | ID: mdl-39036061

ABSTRACT

We have placed a van der Waals homostructure, formed by stacking three two-dimensional layers of WS2 separated by insulating hBN, similar to a multiple-quantum well structure, inside a microcavity, which facilitates the formation of quasiparticles known as exciton-polaritons. The polaritons are a combination of light and matter, allowing laser emission to be enhanced by nonlinear scattering, as seen in prior polariton lasers. In the experiments reported here, we have observed laser emission with an ultralow threshold. The threshold was approximately 59 nW/µm2, with a lasing efficiency of 3.82%. These findings suggest a potential for efficient laser operations using such homostructures.

19.
Mol Plant ; 17(9): 1472-1483, 2024 Sep 02.
Article in English | MEDLINE | ID: mdl-39049493

ABSTRACT

Gene upregulation through genome editing is important for plant research and breeding. Targeted insertion of short transcriptional enhancers (STEs) into gene promoters may offer a universal solution akin to transgene-mediated overexpression while avoiding the drawbacks associated with transgenesis. Here, we introduce an "in locus activation" technique in rice that leverages well-characterized STEs for refined, heritable, and multiplexed gene upregulation. To address the scarcity of potent enhancers, we developed a large-scale mining approach and discovered a suite of STEs that are capable of enhancing gene expression in rice protoplasts. The in locus integration of these STEs into eight rice genes resulted in substantial transcriptional upregulation in the edited plants, with up to 869.1-fold increases in their transcript levels. Employing a variety of STEs, we achieved delicate control of gene expression, enabling the fine-tuning of key phenotypic traits such as plant height. Our approach also enabled efficient multiplexed gene upregulation, with up to four genes activated simultaneously, significantly enhancing the nicotinamide mononucleotide metabolic pathway. Importantly, heritability studies from the T0 to T3 generations confirmed the stable and heritable nature of STE-driven gene activation. Collectively, our work demonstrates that coupled with STE mining, leveraging genome editing for in locus activation and gene upregulation holds great promise to be widely adopted in fundamental plant research and crop breeding.


Subject(s)
CRISPR-Cas Systems , Enhancer Elements, Genetic , Gene Editing , Oryza , Up-Regulation , Oryza/genetics , Up-Regulation/genetics , Enhancer Elements, Genetic/genetics , Gene Editing/methods , Gene Expression Regulation, Plant , Plants, Genetically Modified/genetics
20.
Plant Commun ; : 101040, 2024 Jul 20.
Article in English | MEDLINE | ID: mdl-39001607

ABSTRACT

Understanding the behavior of endogenous proteins is crucial for functional genomics, yet their dynamic characterization in plants presents substantial challenges. Whereas mammalian studies have leveraged in locus tagging with the luminescent HiBiT peptide and genome editing for rapid quantification of native proteins, this approach remains unexplored in plants. Here, we introduce the in locus HiBiT tagging of rice proteins and demonstrate its feasibility in plants. We found that although traditional HiBiT blotting works in rice, it failed to detect two of the three tagged proteins, a result attributable to low luminescence activity in plants. To overcome this limitation, we engaged in extensive optimization, culminating in a new luciferin substrate coupled with a refined reaction protocol that enhanced luminescence up to 6.9 fold. This innovation led to the development of TagBIT (tagging with HiBiT), a robust method for high-sensitivity protein characterization in plants. Our application of TagBIT to seven rice genes illustrates its versatility on endogenous proteins, enabling antibody-free protein blotting, real-time protein quantification via luminescence, in situ visualization using a cross-breeding strategy, and effective immunoprecipitation for analysis of protein interactions. The heritable nature of this system, confirmed across T1 to T3 generations, positions TagBIT as a powerful tool for protein study in plant biology.

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