Genetically engineered mouse models (GEMMs) are crucial for investigating disease mechanisms, developing therapeutic strategies, and advancing fundamental biological research. While CRISPR gene editing has greatly facilitated the creation of these models, existing techniques still present technical challenges and efficiency limitations. Here, we establish a CRISPR-VLP-induced targeted mutagenesis (CRISPR-VIM) strategy, enabling precise genome editing by co-culturing zygotes with virus-like particle (VLP)-delivered gene editing ribonucleoproteins (RNPs) without requiring physical manipulation or causing cellular damage. We generate Plin1- and Tyr-knockout mice through VLP-based SpCas9 or adenine base editor (ABE)/sgRNA RNPs and characterize their phenotype and germline transmission. Additionally, we demonstrate cytosine base editor (CBE)/sgRNA-based C-to-T substitution or SpCas9/sgRNA-based knock-in using VLPs. This method further simplifies and accelerates GEMM generation without specialized techniques or equipment. Consequently, the CRISPR-VIM method can facilitate mouse modeling and be applied in various research fields.

During T-cell-mediated inflammatory responses, T cells are activated upon recognizing specific antigens presented by antigen-presenting cells. This recognition initiates signaling through the TCR and CD28, leading to their activation and subsequent clonal expansion. Within the signaling cascades triggered by TCR and CD28 engagement, the CD28-PI3K pathway serves as a central regulator of metabolic reprogramming in T cells, supporting the biosynthetic needs essential for their effective proliferation. In this study, we found that the regulation of PANK4 plays a role in TCR/CD28-mediated CD4⁺ T-cell proliferation by regulating de novo lipid synthesis. The CD28 signaling pathway negatively regulates PANK4 through direct binding with PDK1, thereby controlling de novo lipid synthesis for CD4⁺ T-cell proliferation. Interestingly, we found that Pank4-deficient CD4⁺ T cells enhance coenzyme A synthesis and glutaminolysis, whereby glutamine contributes carbon for fatty acid synthesis and provides nitrogen for coenzyme A biosynthesis. The regulatory role of PANK4 in CD4⁺ T-cell proliferation was confirmed in models of experimental colitis and influenza A virus infection, where Pank4-deficient CD4⁺ T cells exhibited greater expansion than their wild-type counterparts when co-transferred. Our findings suggest that PANK4 regulation of de novo lipid synthesis is crucial for TCR/CD28-induced CD4⁺ T-cell proliferation and represents a potential target for modulating general CD4⁺ T-cell responses.

Genetically engineered mouse models (GEMMs) are crucial for investigating disease mechanisms, developing therapeutic strategies, and advancing fundamental biological research. While CRISPR gene editing has greatly facilitated the creation of these models, existing techniques still present technical challenges and efficiency limitations. Here, we establish a CRISPR-VLP-induced targeted mutagenesis (CRISPR-VIM) strategy, enabling precise genome editing by co-culturing zygotes with virus-like particle (VLP)-delivered gene editing ribonucleoproteins (RNPs) without requiring physical manipulation or causing cellular damage. We generate Plin1- and Tyr-knockout mice through VLP-based SpCas9 or adenine base editor (ABE)/sgRNA RNPs and characterize their phenotype and germline transmission. Additionally, we demonstrate cytosine base editor (CBE)/sgRNA-based C-to-T substitution or SpCas9/sgRNA-based knock-in using VLPs. This method further simplifies and accelerates GEMM generation without specialized techniques or equipment. Consequently, the CRISPR-VIM method can facilitate mouse modeling and be applied in various research fields.

During T-cell-mediated inflammatory responses, T cells are activated upon recognizing specific antigens presented by antigen-presenting cells. This recognition initiates signaling through the TCR and CD28, leading to their activation and subsequent clonal expansion. Within the signaling cascades triggered by TCR and CD28 engagement, the CD28-PI3K pathway serves as a central regulator of metabolic reprogramming in T cells, supporting the biosynthetic needs essential for their effective proliferation. In this study, we found that the regulation of PANK4 plays a role in TCR/CD28-mediated CD4⁺ T-cell proliferation by regulating de novo lipid synthesis. The CD28 signaling pathway negatively regulates PANK4 through direct binding with PDK1, thereby controlling de novo lipid synthesis for CD4⁺ T-cell proliferation. Interestingly, we found that Pank4-deficient CD4⁺ T cells enhance coenzyme A synthesis and glutaminolysis, whereby glutamine contributes carbon for fatty acid synthesis and provides nitrogen for coenzyme A biosynthesis. The regulatory role of PANK4 in CD4⁺ T-cell proliferation was confirmed in models of experimental colitis and influenza A virus infection, where Pank4-deficient CD4⁺ T cells exhibited greater expansion than their wild-type counterparts when co-transferred. Our findings suggest that PANK4 regulation of de novo lipid synthesis is crucial for TCR/CD28-induced CD4⁺ T-cell proliferation and represents a potential target for modulating general CD4⁺ T-cell responses.

Abstract Heart weight (HW) is a critical parameter in cardiology and mouse research, commonly normalized to body weight (BW) or tibia length (TL) to account for size differences. Ratio-based normalization, however, assumes strict proportionality between variables, an assumption that is rarely tested and may bias group comparisons. We analysed HW, BW, and TL measurements from over 25,000 C57BL/6N wildtype mice generated by the International Mouse Phenotyping Consortium. Sex- and age-stratified analyses were combined with simulation-based modelling to evaluate empirical scaling relationships and the statistical behaviour of ratio-based normalization. Across all age and sex groups, correlations between HW, BW, and TL were negligible to weak, indicating substantial deviations from proportionality. Simulations demonstrated that ratio-based normalization can generate misleading results, including spurious or reversed group differences, when proportionality assumptions are violated. Ratios were consistent with linear and allometric models only under strictly proportional conditions, characterized by regression lines passing through the origin. Linear models with covariate adjustment and allometric scaling provide more robust and biologically meaningful frameworks for organ weight analysis. Ratio-based normalization should be avoided unless key mathematical assumptions are met.

These findings suggest that Ei24 overexpression contributes to improved glucose regulation and healthier aging across multiple organs. By interacting with IGF1R, EI24 may provide a novel mechanism for promoting metabolic and age-related health.

This study offers guidance for designing successful EE programs in laboratory animal research institutions.

Fecal sampling is a widely used, non-invasive method for assessing gut microbiomes across various organisms. However, its suitability for studying the gut microbiome of honey bee ( Apis mellifera ) queens has not been tested. In this study, we evaluated whether fecal microbiomes accurately reflect gut microbiomes in honey bee queens, offering a potential non-destructive approach for microbiome research. We successfully obtained fecal and gut samples from 21 out of 26 queens. Bacterial communities were analyzed using 16S rRNA amplicon sequencing and qPCR. Our results indicate that queen fecal microbiomes closely resemble gut microbiomes, with no significant differences in alpha diversity and only minor differences in specific bacterial taxa. Beta diversity analyses revealed that within-pair microbiomes (i.e., gut vs. feces from the same queen) were significantly more similar than between-pair comparisons. Additionally, qPCR analyses revealed a strong positive correlation between bacterial abundances in fecal and gut samples, further supporting the use of feces as a proxy for gut microbiome composition. While promising, fecal collection from queens can sometimes be challenging. In our study, we were unable to collect feces from five queens, and these individuals lacked stored fecal material upon inspection of dissected guts. Nonetheless, our findings suggest that fecal sampling can be a useful, non-invasive method for studying honey bee queen microbiomes, enabling longitudinal assessments without compromising colony stability.

SARS-CoV-2 infection indirectly leads to profound and post-acute COVID-19 alterations in the stomach at the cellular and molecular levels, resulting in adverse outcomes following co-infection with SARS-CoV-2 and Hpylori. Our results show that 2 prevalent pathogens of humans elicit a negative synergistic effect and provide evidence of the risk of severe chronic gastritis in the post-COVID-19 era.