Biology subjects

Wang, X.-C.

Publications and source records attributed to Wang, X.-C..

4 recordsLinked to original sources

bioRxiv2025

Unveiling the Role of PHF20 in TBC1D4-Mediated Glucose Uptake During Toxoplasma gondii Infection

BackgroundToxoplasma gondii relies on host cell nutrients, especially glucose, to support its intracellular growth. However, the mechanisms by which it enhances host glucose metabolism remain incompletely understood. MethodsWe used fluorescence-based glucose uptake assays, flow cytometry, metabolic flux analysis, and gene silencing in ARPE-19 cells, along with transgenic mouse models, to explore how T. gondii manipulates host glucose utilization and whether this promotes parasite proliferation. ResultsT. gondii infection significantly enhanced host glucose uptake in a dose-dependent manner and promoted GLUT4 translocation to the plasma membrane. Notably, extracellular acidification rate (ECAR) assays demonstrated a marked increase in glycolytic activity following infection. Mechanistically, we identified that the PI3K/AKT signaling pathway mediates the phosphorylation and transcriptional upregulation of TBC1D4, a Rab-GAP protein essential for GLUT4 trafficking. Genetic silencing of TBC1D4 impaired both glucose uptake and parasite replication. Furthermore, we uncovered a regulatory mechanism involving AKT-dependent nuclear signaling that modulates TBC1D4 transcription. In vivo experiments using Phf20 transgenic mice confirmed increased susceptibility to T. gondii and elevated glucose metabolic responses. ConclusionsOur study reveals that T. gondii hijacks a host PI3K/AKT-TBC1D4 axis to enhance glucose uptake and glycolysis, thereby ensuring metabolic support for its proliferation. These findings highlight host glucose metabolism as a potential therapeutic target for controlling toxoplasmosis.

cell biology↗

bioRxiv2023

Geranylgeranyl pyrophosphate (GGPP) is associated with hepatic lipid accumulation and insulin resistance in MAO by prenylating Perilipin4

Metabolically Abnormal Obesity (MAO) is characterized by hepatic steatosis and type 2 diabetes (T2D), in contrast to Metabolically Healthy Obesity (MHO). In this study, we investigated the role of hepatic geranylgeranyl pyrophosphate (GGPP), a metabolite of the mevalonate (MVA) pathway, in regulating the differences in lipid metabolism between MAO and MHO. Our findings revealed that GGPP levels were significantly elevated in individuals with MAO, and deficiency of GGPP in the liver ameliorated the defects associated with MAO. Furthermore, we discovered that the prenylation of the lipid droplet-associated protein Perilipin 4 by GGPP enhances the formation of large lipid droplets, thereby exacerbating hepatic lipid accumulation and insulin resistance. Notably, the inhibitor DGBP, targeting the GGPP synthase Ggpps, effectively attenuated the traits of MAO, offering novel insights into the treatment of this condition.

pathology↗

bioRxiv2021

MS2 RNA aptamer enhances prime editing in rice

Prime editing is a universal and very promising precise genome editing technology. However, optimization of prime editor (PE) from different aspects remains vital for its use as a routine tool in plant basic research and crop molecular breeding. In this report, we tested MS2-based prime editor (MS2PE). We fused the M-MLV reverse transcriptase (RT) gene variant to the MS2 RNA binding protein gene, MCP, and allowed the MCP-RT fusion gene to co-express with the SpCas9 nickase gene, SpCas9H840A, and various engineered pegRNAs harboring MS2 RNA (MS2pegR). Compared with control PEs, MS2PEs significantly enhanced editing efficiency at four of six targets in rice protoplasts, and achieved 1.2[~]10.1-fold increase in editing efficiency at five of six targets in transgenic rice lines. Furthermore, we tested total 22 different MS2pegR scaffolds, 3 RT variants or genes, 2 MCP variants, and various combinations of the Cas9 nickase, RT, and MCP modules. Our results demonstrated an alternative strategy for enhancing prime editing.

molecular biology↗

bioRxiv2017

High-frequency off-target mutagenesis induced by CRISPR/Cas9 in Arabidopsis and its prevention by improving specificity of the tools

Specificity of CRISPR/Cas9 tools has been a major concern along with the reports of their successful applications. We report unexpected observations of high frequency off-target mutagenesis induced by CRISPR/Cas9 in T1 Arabidopsis mutants although the sgRNA was predicted to have a high specificity score. We also present evidence that the off-target effects were further exacerbated in the T2 progeny. To prevent the off-target effects, we tested and optimized two strategies in Arabidopsis, including introduction of a mCherry cassette for a simple and reliable isolation of Cas9-free mutants and the use of highly specific mutant SpCas9 variants. Optimization of the mCherry vectors and subsequent validation found that fusion of tRNA with the mutant rather than the original sgRNA scaffold significantly improves editing efficiency. We then examined the editing efficiency of eight high-specificity SpCas9 variants in combination with the improved tRNA-sgRNA fusion strategy. Our results suggest that highly specific SpCas9 variants require a higher level of expression than their wild-type counterpart to maintain high editing efficiency. Additionally, we demonstrate that T-DNA can be inserted into the cleavage sites of CRISPR/Cas9 targets with high frequency. Altogether, our results suggest that in plants, continuous attention should be paid to off-target effects induced by CRISPR/Cas9 in current and subsequent generations, and that the tools optimized in this report will be useful in improving genome editing efficiency and specificity in plants and other organisms.

molecular biology↗