adenoviral cre  (Vector Biolabs)

Journal: Biochemistry and Biophysics Reports

Article Title: LIN28-mediated gene regulatory loops synchronize transitions throughout organogenesis

doi: 10.1016/j.bbrep.2025.102226

Figure Lengend Snippet: Transcriptional regulation of LIN28A and LIN28B by developmental transcription factors. A) Schematic of transcription factors that orchestrate germ layer formation and early embryonic lineage specification and differentiation. B) Top: Schematic of morphogen-induced lung/endoderm formation from iPSCs through initiation of lung progenitors in vitro. Bottom: Gene expression analysis of single cell RNA-sequencing of roughly 10,000 cells (close 1200 for each day) throughout morphogen-directed lung endoderm formation from iPSCs. Data from manuscript (Ori et al., 2023). C) Fold change in gene expression of LIN28A and LIN28B upon infection of embryonic lung fibroblast WI-38, with adenoviral constructs: GFP, FOXA2, NKX2-1, SOX2 and SOX9 for 48–72 h before lysing in Trizol. Dots on the graph representative of n = 2 independent experiments in quadruplicate. D) Schematic of development of endoderm differentiation. Transient transfection of txrn factors into HEK293 with construct stably expressing dual luciferase construct; firefly and Renilla luciferase. Validation of a subset of factors that regulated human LIN28A and LIN28B promoter/enhancer sequences during screening experiments in panels B) and C) during E) pluripotency and F) endoderm/lung development. G) progenitor specification. Graphs in E), F) and G) are representative of n = 2 independent validation experiments out of 3. P values for C) were determined using 2-way ANOVA of normal distribution. P values of significance: In the order of left to right for panel C: LIN28A – FOXA2- ∗0.024, NKX2-1- ∗0.035, SOX9- ∗0.021; LIN28B – NKX2-1- ∗0.021, ∗SOX9- 0.021 P values for graphs E), F) and G) were determined using 1-way ANOVA of normal distribution in E) and 1-way ANOVA of lognormal distribution for F) and G). P values of significance: In the order of left to right for each panel: E)∗ 0.0375, ∗∗ 0.0021; ∗ 0.0184, ∗0.0321 F)∗ 0.0434, ∗0.0325; ∗0.0191, ∗0.0477 G)∗∗0.0097,∗ 0.0112; ∗∗0.0021, ∗∗0.0078; ∗∗ 0.0036, ∗∗0.0079; ∗∗0.0072, ∗0.0160. Schematics A), B), D) were made using BioRender.

Article Snippet: Lin28a/b were knocked out post-neurosphere formation via adenoviral infection with Ad-Cre-GFP adenovirus (Vector Biolabs, 1700) or Ad-GFP adenovirus (Vector Biolabs, 1060).

Techniques: In Vitro, Gene Expression, RNA Sequencing, Infection, Construct, Transfection, Stable Transfection, Expressing, Luciferase, Biomarker Discovery

Journal: The Journal of physiology

Article Title: Diet-induced obesity in mice increases carotid body chemosensitivity via the leptin-TRPM7 pathway

doi: 10.1113/JP288722

Figure Lengend Snippet: Confocal images showing Ad-Cre-GFP expression, suggesting successful transfection of the Ad-Cre-GFP viruses into the CB of diet-induced obese (DIO) Trpm7 flox mice ( N = 2). B , summary data showing relative expression ratio (RER) of Trmp7 mRNA in the carotid bodies of lean Trmp7 flox mice treated with control Ad-GFP ( N = 6) and Ad-Cre-GFP viruses ( N = 4). There was a significant decrease in the Trpm7 mRNA expression in the Ad-Cre-GFP group ( P = 0.002; Mann–Witney U test).

Article Snippet: Adenoviral vectors Ad-Cre-GFP containing Cre-recombinase tagged with green fluorescent protein (GFP, 3.3 × 10 12 VP/ml) and control Ad-GFP (3.3 × 10 12 VP/ml) were from Vector Biolabs (Malvern, PA, USA).

Techniques: Expressing, Transfection, Control

Journal: The Journal of physiology

Article Title: Diet-induced obesity in mice increases carotid body chemosensitivity via the leptin-TRPM7 pathway

doi: 10.1113/JP288722

Figure Lengend Snippet: A and B are representative illustrations of ex vivo CSN responses from diet-induced obese Trpm7 flox mice transfected into the CB with Ad-Cre-GFP and control Ad-GFP viruses, respectively. CSN responses were normalized to activity in hyperoxia (Hypx, PO 2 = 200 Torr, represented by the horizontal dashed line). Genetic knockdown of Trpm7 receptors completely abolished the stimulatory effect of leptin on CSN activity ( A ), whereas the leptin’s effect was intact when the Trpm7 channels were functional. B , the P -values for the effect of Trpm7 knockdown are shown in the figures C and D . C , summary data of basal CSN activity: normoxia (Nx) + Ad-Cre-GFP vs . Nx + leptin + Ad-Cre-GFP ; P = 0.148 ( N = 5), Nx + Ad-GFP vs . Nx + leptin + Ad-GFP ; P = 0.001 ( N = 5). D , the hypoxic chemoreflex represented as ΔCSN activity (Hx–Nx): Ad-Cre-GFP vs. Ad-Cre-GFP + leptin; P = 1.000, Ad-GFP vs. Ad-GFP + leptin; P < 0.001, Ad-Cre-GFP + leptin vs. Ad-Cre-GFP + leptin + FTY720; P = 0.456; Ad-GFP + leptin vs. Ad-GFP + leptin + FTY720; P = 0.001 ( N = 5). There was a statistically significant interaction between the Ad-GFP vs. Ad-Cre-GFP treatment groups and leptin treatment at hypoxic condition; P = 0.006. Data were compared using two-way repeated-measures ANOVA: no leptin vs . leptin vs . leptin + FTY720 within subjects, Ad-Cre-GFP vs . control Ad-GFP viruses between subjects. Sphericity was assessed using Mauchly’s test in both C and D and found not to be violated. All values are expressed as box-and-whisker plots (median, 25%–75% centiles and minimum and maximum values).

Article Snippet: Adenoviral vectors Ad-Cre-GFP containing Cre-recombinase tagged with green fluorescent protein (GFP, 3.3 × 10 12 VP/ml) and control Ad-GFP (3.3 × 10 12 VP/ml) were from Vector Biolabs (Malvern, PA, USA).

Techniques: Knockdown, Activity Assay, Ex Vivo, Transfection, Control, Functional Assay, Whisker Assay