Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 2. Inhibition of Rosea1 activity in antirrhinum fl ower buds using transient RNA interference (RNAi). Petals of buds cul- tured in vitro are shown 12–17 days after the biolistic introduction of the plasmid for Rosea1 RNAi inhibition. The adaxial ( left ) and abaxial ( right ) epidermis of the same region of one petal are shown. The same pattern of inhibition on both surfaces demonstrates that the silencing signal was transmitted from the bombarded abaxial epidermis to the adaxial epidermis.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Inhibition, Activity Assay, In Vitro, Plasmid Preparation

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 5. The transfection ef fi ciency of biolistics bombardment of plasmid gWIZ- luc coated gold particles at different particle size and helium pressure using Human Embryonic Kidney (HEK) 293 cells (4 × 10 5 cells/well, 12-well plate) at 48 h after bombardment transfection. Gold particles : 0.6, 1.0, and 1.6 μ m, Pressure: 50, 75, 100, 150, 200 psi, Plasmid DNA : gWIZ- luc , Dose : 0.5 mg gold particles/1 μ g DNA/shot (cartridge). Transfection ef fi ciency compared with negative control (untransfected cells) and positive control (Lipofectamine TM 2000 as transfection reagent).

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Transfection, Plasmid Preparation, Negative Control, Positive Control

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 3. A myocyte transfected with an expression vector encoding mCherry. Freshly isolated rat ventricular myocytes were transfected with pcDNA3-mCherry coated 0.6 m m gold beads at a pressure of 90 psi. After bombardment, the myocytes were cultured for 24 h in Supplemented M199 media then visualized using an Olympus Fluoview 1000 laser scanning confocal microscope. The transfected myocyte, visible at upper left in each frame, glows brightly in the red ( grey in the greyscale image). Also visible is a nearby untransfected myocyte and normal detritus resulting from the biolistic transfec- tion. Left , bright fi eld image. Middle , fl uorescence image. Right , combined bright fi eld and fl uorescence image.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Transfection, Expressing, Plasmid Preparation, Isolation, Cell Culture, Microscopy

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 1. Low pressure gene gun. ( a ) Schematic drawing depicting the features of the gene gun. Plasmid is loaded into the upper hole. When pressing the trigger, the helium fl ow travels from the inside of the spray nozzle to the outside and the pressure difference results in supersonic fl ow to carry particles-coated plasmids or the plasmid solution to target cells. ( b ) Shaved abdominal skin of mice. ( c ) Photograph showing the upper hole of the low pressure gene gun. ( d ) Loading of plas- mid DNA into the upper hole of the low pressure gene gun. ( e ) Biolistic DNA delivery onto a mouse. One person holds the mouse and another person holds the gene gun which was loaded with plasmid DNA. ( f ) Gene gun bombardment on the shaved abdomen.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Plasmid Preparation

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 2. Evaluation of ef fi ciency of plasmid DNA delivery with the low pressure gene gun. ( a ) Empty plasmid vector (pGL3- Basic), 10 m g luciferase plasmid without particles and 1 m g luciferase plasmid coated on gold particles, respectively, was bombarded to C57BL/6 mice. After 48 h, luciferase activity was detected by in vivo image system. ( b ) Quanti fi cation of luciferase activity. 48 h after biolistic transfection, the skin from the plasmid-bombarded mice was homogenized. The luciferase activity of the skin lysate was determined.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Plasmid Preparation, Luciferase, Activity Assay, In Vivo, Transfection

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 2. Development of different Th cell subsets in draining lymph nodes following biolistic transfection with pFascin- b Gal or pCMV- b Gal. BALB/c mice ( n = 4) were immunized by fi ve gene gun-mediated immunizations with pFascin- b Gal and pCMV- b Gal, respectively, in weekly intervals. Fifty-six days after the fi rst immunization spleen and draining ingui- nal/axillary lymph nodes cells were prepared and cultured (5 × 10 6 /well) in quadruplicates with or without 25 m g/mL b Gal as antigen. After 72 h culture supernatants were pooled and the amount of IFN- g ( open bars ) and IL-5 ( closed bars ) was determined.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Transfection, Cell Culture

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 1. Qualitative differences in the IgG pro fi le following biolistic transfection with pFascin- b Gal or pCMV- b Gal. BALB/c mice ( n = 4) were immunized by three gene gun-mediated immunizations with pFascin- b Gal and pCMV- b Gal, respectively, in weekly intervals. Thirty- fi ve days after the fi rst immunization sera were collected and levels of b Gal-speci fi c IgG1 ( open bars ) and IgG2a ( closed bars ) were determined.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Transfection

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 3. Recruitment of IFN- g -producing CD8 + effector T cells following biolistic transfec- tion with pFascin- b Gal or pCMV- b Gal. BALB/c mice ( n = 4) were immunized by three gene gun-mediated immunizations with pFascin- b Gal and pCMV- b Gal, respectively, in weekly intervals. Twenty-eight days after the fi rst immunization numbers of IFN- g - producing CD8 + effector T cells among spleen cells were determined following incuba- tion of splenocytes without ( open bars ) or with ( closed bars ) 0.1 m g/mL b Gal-speci fi c peptide TPHPARIGL for 22 h.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques:

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 3. T . cruzi infectious challenge of immunized mice. ( a ) Balb/c mice are more susceptible than C57Bl/6 mice to challenge with TCT (i.p.). Plots show the survival of mice inoculated with the indicated dose of parasites. ( b ) Parasitemia after T . cruzi challenge of biolistic DNA immunized (CRP and TcPRAC) vs. vector control mice. ( c ) Amastigote parasite nest ( arrow ) in heart tissue of T . cruzi challenged mouse (H&E stain, 800× magni fi cation).

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Plasmid Preparation, Control, Staining

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 1. Inhibition of IgE production by prophylactic gene gun-mediated vaccination. BALB/c mice ( n = 4) were vaccinated every week by a total of fi ve biolistic transfections with pCMV- β Gal or a non-encoding control vector or were left unvaccinated. Subsequently mice were sensitized by six intraperitoneal injections of 1 μ g β Gal adsorbed to aluminum hydroxide as adjuvant at intervals of 2 weeks. Four days after the sixth application of β Gal, levels of β Gal-speci fi c IgE in sera were determined by ELISA. Data represent mean β Gal- speci fi c IgE titer ± SD. ** P < 0.01.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Inhibition, Transfection, Control, Plasmid Preparation, Adjuvant, Enzyme-linked Immunosorbent Assay

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 2. Reduction of pulmonary eosinophilic in fi ltration by prophylactic gene gun-mediated vaccination. BALB/c mice ( n = 4) were vaccinated every week by a total of three biolistic transfections with pCMV- β Gal or a non-encoding control vector. Subsequently vaccinated mice as well as untreated control mice were sensitized at a 10 day interval by two intra- peritoneal injections of 1 μ g β Gal adsorbed to aluminum hydroxide as adjuvant. One week later all mice were challenged with antigen on three consecutive days by intranasal appli- cation of 50 μ g β Gal. Twenty-four hour after the last challenge BAL was performed and BAL cells were differentiated using Diff-Quick staining. M monocyte/macrophage; L lym- phocyte; E eosinophil; N neutrophil.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Transfection, Control, Plasmid Preparation, Adjuvant, Diff-Quik, Staining

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 3. Measurement of airway reactivity after prophylactic gene gun-mediated vaccina- tion. BALB/c mice ( n = 4) were vaccinated every week by a total of three biolistic transfec- tions with pCMV- β Gal or a non-encoding control vector. Subsequently vaccinated mice as well as untreated control mice were sensitized at a 10 day interval by two intraperitoneal injections of 1 μ g β Gal adsorbed to aluminum hydroxide as adjuvant. One week later (all) mice were challenged with antigen or saline on three consecutive days by intranasal application of 50 μ g β Gal, a separate control group of sensitized mice was treated with PBS. Twenty-four hours after the last challenge airway reactivity was measured by nonin- vasive plethysmography in response to increasing doses of methacholine (MCh) as enhanced pause (Penh). Data represent means. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Key words: Caenorhabditis elegans , Biolistic transformation , Bio-Rad Biolistic PDS-1000/He system , Hepta adapter , unc-119 ( ed3 ) Caenorhabditis elegans is a small, free living round worm, which feeds on bacteria and fungi in soil.

Techniques: Control, Plasmid Preparation, Adjuvant, Saline