ad cmv luciferase adenovirus infection  (Vector Biolabs)

Journal: Bio-protocol

Article Title: Temporally and Spatially Controlled Age-Related Prostate Cancer Model in Mice

doi: 10.21769/BioProtoc.5144

Figure Lengend Snippet: (A) Anesthetize the mouse. (B) Cover the mouse under a semi-transparent food wrap with the surgical field exposed (outlined with dashed lines). (C) Create an abdominal incision using a scalpel. (D) Expose the genitourinary (GU) bloc. The cartoon and the magnified photo of the mouse's GU bloc demonstrate the anatomy of seminal vesicles, anterior prostate tissue, and the urinary bladder. L, left. R, right. AP, anterior prostate. SV, seminal vesicles. (E) Inject Ad-Cre-Luc viral particles into APs on both sides (this panel only shows injection into the right AP). Ad-Cre-Luc, adenovirus viral particle driving the co-expression of Cre recombinase and luciferase. (F) Close the incision. (G) Let the mouse recover from anesthesia. (H) Anesthetize the mouse. (I) Peritoneal injection of D-luciferin. (J) Detect the infection of Ad-Cre-Luc using luciferase-based imaging. (K) Let the mouse recover from anesthesia.

Article Snippet: Luciferase adenovirus (Ad-CMV-Luc) (VECTOR BIOLABS, catalog number: 1000); this adenovirus vector delivers luciferase.

Techniques: Injection, Expressing, Luciferase, Infection, Imaging

Journal: Bio-protocol

Article Title: Temporally and Spatially Controlled Age-Related Prostate Cancer Model in Mice

doi: 10.21769/BioProtoc.5144

Figure Lengend Snippet: Representative image of a mouse with bilateral anterior prostate (AP) infection of Adenovirus that drives the expression of the reporter gene, luciferase. The image was acquired five days after the surgery. The color-coded scale bars on the right side indicate signal intensities of the luciferase-catalyzed reaction in the image on the left.

Article Snippet: Luciferase adenovirus (Ad-CMV-Luc) (VECTOR BIOLABS, catalog number: 1000); this adenovirus vector delivers luciferase.

Techniques: Infection, Expressing, Luciferase

Journal: Bio-protocol

Article Title: Temporally and Spatially Controlled Age-Related Prostate Cancer Model in Mice

doi: 10.21769/BioProtoc.5144

Figure Lengend Snippet: This figure is adapted from the authors' published article, A Novel Controlled PTEN-Knockout Mouse Model for Prostate Cancer Study (Figure 2, Panels A–C). Frontiers in Molecular Biosciences [8]. (A) IHC staining to detect Cre recombinase in Pten LoxP/LoxP (L/L) mice's anterior prostate (AP) from different age groups that are infected with different adenovirus vectors. Ad-CMV-Luc, adenovirus vector that expresses luciferase using a cytomegalovirus (CMV) promoter. Green arrows indicate the basal cells. Green dash lines outline the stroma. Green triangles point toward stroma cells. Pten , phosphatase and tensin homolog deleted on chromosome 10. Ad-Cre-Luc, adenovirus vector that co-expresses luciferase and Cre recombinase (also using the CMV promoter). (B) IHC staining to detect Pten and downstream components of the PI3K/mTOR pathways in Pten L/L mice's AP that were infected with Ad-CMV-Luc and Ad-Cre-Luc viral vectors. Red arrows indicate Pten-, P-Akt-, P-S6- , or P-4E-BP1- positive cells. P-S6 , phospho-S6 ribosomal protein. P-4E-BP1 , phospho-eukaryotic translation initiation factor 4E-binding protein 1. (C) Serial IHC staining to detect Pten and downstream components of the PI3K/mTOR pathways in Pten L/L mice's AP at 0, 4, 8, and 16 weeks from the surgical delivery of the viral vectors. Red arrows indicate Pten -negative cells with activated P-S6 and P-4E-BP1 .

Article Snippet: Luciferase adenovirus (Ad-CMV-Luc) (VECTOR BIOLABS, catalog number: 1000); this adenovirus vector delivers luciferase.

Techniques: Knock-Out, Immunohistochemistry, Infection, Plasmid Preparation, Luciferase, Binding Assay

Journal: bioRxiv

Article Title: APOBEC3 activity promotes the survival and evolution of drug-tolerant persister cells during acquired resistance to EGFR inhibitors in lung cancer

doi: 10.1101/2023.07.02.547443

Figure Lengend Snippet: A. qRT-PCR analysis showing A3A and A3B expression in PC9 cells following treatment with gefitinib or osimertinib over the course of 14 days. Error bars represent SEM of three technical replicates. B. Schematic of Cre-inducible APOBEC3B expression. C. qRT-PCR analysis showing A3B expression in PC9 cells following infection with Cre recombinase. Error bars represent SEM of three technical replicates. * indicates p<0.05. BT474 and SKBR3 cells are shown as controls. D. Western blot showing protein expression of HA-tagged A3B in PC9 cells following infection with Cre recombinase. E. In vitro deaminase activity assay in PC9 cells following infection with Cre recombinase. BT474 and SKBR3 cells are shown as controls. % deamination was calculated as described in Methods. F. Quantification of % deamination in two replicates of control PC9 cells (-Cre) and five replicates of A3B-expressing PC9 (+Cre). % deamination was calculated as described in Methods. Unpaired t-test was performed to determine statistical significance. ** indicates p<0.005. G. Growth curves for PC9 cells expressing A3B (+Cre) or control cells (-Cre). Error bars represent SEM of two biological replicates. Two-way ANOVA was performed to determine statistical significance. ns = not significant

Article Snippet: After 24 hr, AdenoCre recombinase (Vector Biolabs) was added to the media at an MOI of 1000.

Techniques: Quantitative RT-PCR, Expressing, Infection, Western Blot, In Vitro, Activity Assay, Control

Journal: bioRxiv

Article Title: APOBEC3 activity promotes the survival and evolution of drug-tolerant persister cells during acquired resistance to EGFR inhibitors in lung cancer

doi: 10.1101/2023.07.02.547443

Figure Lengend Snippet: A. qRT-PCR analysis showing A3A and A3B expression in PC9 cells following treatment with gefitinib or osimertinib over the course of 14 days. Error bars represent SEM of three technical replicates. B. Schematic of Cre-inducible APOBEC3B expression. C. qRT-PCR analysis showing A3B expression in PC9 cells following infection with Cre recombinase. Error bars represent SEM of three technical replicates. * indicates p<0.05. BT474 and SKBR3 cells are shown as controls. D. Western blot showing protein expression of HA-tagged A3B in PC9 cells following infection with Cre recombinase. E. In vitro deaminase activity assay in PC9 cells following infection with Cre recombinase. BT474 and SKBR3 cells are shown as controls. % deamination was calculated as described in Methods. F. Quantification of % deamination in two replicates of control PC9 cells (-Cre) and five replicates of A3B-expressing PC9 (+Cre). % deamination was calculated as described in Methods. Unpaired t-test was performed to determine statistical significance. ** indicates p<0.005. G. Growth curves for PC9 cells expressing A3B (+Cre) or control cells (-Cre). Error bars represent SEM of two biological replicates. Two-way ANOVA was performed to determine statistical significance. ns = not significant

Article Snippet: 24 hr later, adenovirus expressing Cre recombinase (Vector Biolabs) was added to the media at an MOI of 1000.

Techniques: Quantitative RT-PCR, Expressing, Infection, Western Blot, In Vitro, Activity Assay, Control

Journal: Bioengineering

Article Title: Development of Adenovirus Containing Liposomes Produced by Extrusion vs. Homogenization: A Comparison for Scale-Up Purposes

doi: 10.3390/bioengineering9110620

Figure Lengend Snippet: Comparative in vivo biodistribution of Ad liposomes manufactured by extrusion and homogenization techniques: ( A ) The most representative IVIS images of mice IT injected with unencapsulated AdLuc (n = 5), Ex Df + AdLuc (n = 5), and HMG Df + AdLuc (n = 5) at 1.4 × 10 8 PFU. Both dorsal and ventral sides of each mouse were imaged after 5 days. Red circles highlight injected tumors. ( B ) Average radiance of tumors for Ex Df + AdLuc (n = 5) (red box), HMG Df + AdLuc (n = 5) (blue box), and unencapsulated AdLuc (n = 5) (yellow box) injected mice revealed enhanced transduction of tumors with both Ex Df + AdLuc and HMG Df + AdLuc. Performance of Ex Df + AdLuc, and HMG Df + AdLuc was similar ( p value: ns = not significant). Radiance < 10,000 p/s/cm 2 /sr (shaded region) is considered background radiance ( p value: ns = not significant, # = 0.1204 ## = 0.1345). ( C ) For each mouse the ratio of total tumor signal to liver signal demonstrated that Ex Df + AdLuc (red box) and HMG Df + AdLuc (blue box) reduced off-tumor transduction by approximately 4-folds compared to the unencapsulated AdLuc (yellow box) ( p value: ns = not significant, # = 0.0533 ## = 0.0703).

Article Snippet: Ad-Luciferase (AdLuc) was purchased from Vector BioLabs (Catalog # 1000).

Techniques: In Vivo, Liposomes, Homogenization, Injection, Transduction