Comparison of In Vitro Performances of Nanorod and Nanofiber Polyplexes Prepared from Plasmid DNA and Poly(L-lysine) Terminally Bearing Multi-Arm PEG

2020 ◽  
pp. 13-21
Author(s):  
Ryuta Aono ◽  
Kenta Nomura ◽  
Eiji Yuba ◽  
Atsushi Harada
Keyword(s):  
Plasmid Dna

Evaluation of biocompatibility and efficiency of plasmid DNA delivery by gene-activated hydrogels in vitro

2019 ◽  
Vol XIV (2) ◽  
Author(s):  
I.Y. Bozo ◽  
A.A. Titova ◽  
M.N. Zhuravleva ◽  
A.I. Bilyalov ◽  
M.O. Mavlikeev ◽  
...  
Keyword(s):  
Plasmid Dna ◽  
Dna Delivery

Multifunctional self-assembled cationic peptide nanostructures efficiently carry plasmid DNA in vitro and exhibit antimicrobial activity with minimal toxicity

2014 ◽  
Vol 2 (30) ◽  
pp. 4848-4861 ◽  
Author(s):  
Santosh Yadav ◽  
Manohar Mahato ◽  
Rajiv Pathak ◽  
Diksha Jha ◽  
Bipul Kumar ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Plasmid Dna ◽  
Cationic Peptide ◽  
Minimal Toxicity ◽  
Amphiphilic Peptide ◽  
Self Assembled ◽  
Multifunctional Properties

An amphiphilic peptide–aminoglycoside (Pep–Neo) conjugate has been synthesized, self-assembled into nanostructures and evaluated for its multifunctional properties.

scholarly journals Plasmid DNA initiates replication of yellow fever vaccine in vitro and elicits virus-specific immune response in mice

Virology ◽  
2014 ◽  
Vol 468-470 ◽  
pp. 28-35 ◽  
Author(s):  
Irina Tretyakova ◽  
Brian Nickols ◽  
Rachmat Hidajat ◽  
Jenny Jokinen ◽  
Igor S. Lukashevich ◽  
...  
Keyword(s):  
Immune Response ◽  
Yellow Fever ◽  
Plasmid Dna ◽  
Specific Immune Response ◽  
Yellow Fever Vaccine

scholarly journals Intravenous Delivery of piggyBac Transposons as a Useful Tool for Liver-Specific Gene-Switching

2018 ◽  
Vol 19 (11) ◽  
pp. 3452 ◽  
Author(s):  
Shingo Nakamura ◽  
Masayuki Ishihara ◽  
Satoshi Watanabe ◽  
Naoko Ando ◽  
Masato Ohtsuka ◽  
...  
Keyword(s):  
Plasmid Dna ◽  
Chain Gene ◽  
Specific Gene ◽  
Chromosomal Integration ◽  
Hepatic Disorder ◽  
Murine Liver ◽  
A Chain ◽  
Gene Switching

Hydrodynamics-based gene delivery (HGD) is an efficient method for transfecting plasmid DNA into hepatocytes in vivo. However, the resulting gene expression is transient, and occurs in a non-tissue specific manner. The piggyBac (PB) transposon system allows chromosomal integration of a transgene in vitro. This study aimed to achieve long-term in vivo expression of a transgene by performing hepatocyte-specific chromosomal integration of the transgene using PB and HGD. Using this approach, we generated a novel mouse model for a hepatic disorder. A distinct signal from the reporter plasmid DNA was discernible in the murine liver approximately two months after the administration of PB transposons carrying a reporter gene. Then, to induce the hepatic disorder, we first administered mice with a PB transposon carrying a CETD unit (loxP-flanked stop cassette, diphtheria toxin-A chain gene, and poly(A) sites), and then with a plasmid expressing the Cre recombinase under the control of a liver-specific promoter. We showed that this system can be used for in situ manipulation and analysis of hepatocyte function in vivo in non-transgenic (Tg) animals.

scholarly journals Two Antibody-Guided Lactic-co-Glycolic Acid-Polyethylenimine (LGA-PEI) Nanoparticle Delivery Systems for Therapeutic Nucleic Acids

2021 ◽  
Vol 14 (9) ◽  
pp. 841
Author(s):  
Jian-Ming Lü ◽  
Zhengdong Liang ◽  
Dongliang Liu ◽  
Bin Zhan ◽  
Qizhi Yao ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Plasmid Dna ◽  
Glycolic Acid ◽  
Growth Factor Receptor ◽  
Green Fluorescence Protein ◽  
Active Targeting ◽  
Single Chain ◽  
Targeting Delivery

We previously reported a new polymer, lactic-co-glycolic acid-polyethylenimine (LGA-PEI), as an improved nanoparticle (NP) delivery for therapeutic nucleic acids (TNAs). Here, we further developed two antibody (Ab)-conjugated LGA-PEI NP technologies for active-targeting delivery of TNAs. LGA-PEI was covalently conjugated with a single-chain variable fragment antibody (scFv) against mesothelin (MSLN), a biomarker for pancreatic cancer (PC), or a special Ab fragment crystallizable region-binding peptide (FcBP), which binds to any full Ab (IgG). TNAs used in the current study included tumor suppressor microRNA mimics (miR-198 and miR-520h) and non-coding RNA X-inactive specific transcript (XIST) fragments; green fluorescence protein gene (GFP plasmid DNA) was also used as an example of plasmid DNA. MSLN scFv-LGA-PEI NPs with TNAs significantly improved their binding and internalization in PC cells with high expression of MSLN in vitro and in vivo. Anti-epidermal growth factor receptor (EGFR) monoclonal Ab (Cetuximab) binding to FcBP-LGA-PEI showed active-targeting delivery of TNAs to EGFR-expressing PC cells.

scholarly journals Osteogenic differentiation as a result of BMP-2 plasmid DNA based gene therapy in vitro and in vivo

2011 ◽  
Vol 21 ◽  
pp. 230-242 ◽  
Author(s):  
F Wegman ◽  
◽  
A Bijenhof ◽  
L Schuijff ◽  
FC Öner ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Osteogenic Differentiation ◽  
Plasmid Dna

An attempt at sperm-mediated gene transfer in mice and chickens

1991 ◽  
Vol 71 (2) ◽  
pp. 287-291 ◽  
Author(s):  
J. S. Gavora ◽  
B. Benkel ◽  
H. Sasada ◽  
W. J. Cantwell ◽  
P. Fiser ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Gene Transfer ◽  
Transgenic Mice ◽  
Key Words ◽  
Plasmid Dna ◽  
Dna Transfer ◽  
Laboratory Mice ◽  
Vitro Fertilization ◽  
Bacterial Plasmid

Experiments were carried out to transform laboratory mice and domestic chickens by use of sperm incubated with bacterial plasmid DNA. Following demonstration of "uptake" of such DNA by sperm of both species, attempts were made to replicate a previously published procedure (Lavitrano et al. 1989, Cell 57: 717–723) for producing transgenic mice through in vitro fertilization (IVF). Also, female mice and hens were inseminated (AI) with sperm which had been incubated in a DNA solution. Such incubation did not influence the fertility or hatchability of the hens' eggs. However, no transformed progeny were detected among 45 mice produced by IVF or among 69 mice and 470 chickens produced by AI. Key words: Sperm-mediated DNA transfer, mice, chickens

scholarly journals Cre/LoxP-HBV plasmids generating recombinant covalently closed circular DNA genome upon transfection

2020 ◽  
Author(s):  
Robert L. Kruse ◽  
Xavier Legras ◽  
Mercedes Barzi
Keyword(s):  
Plasmid Dna ◽  
Plasmid Stability ◽  
Intron Insertion ◽  
Circular Dna ◽  
Immunodeficient Mice ◽  
New Therapies ◽  
Covalently Closed Circular Dna ◽  
Dna Silencing

AbstractNew therapies against hepatitis B virus (HBV) require the elimination of covalently closed circular DNA (cccDNA), the episomal HBV genome. HBV plasmids containing an overlength 1.3-mer genome and bacterial backbone (pHBV1.3) are used in many different models, but do not replicate the unique features of cccDNA. Since the stable cccDNA pool is a barrier to HBV eradication in patients, we developed a recombinant circular HBV genome (rcccDNA) to mimic the cccDNA using Cre/LoxP technology. We validated four LoxP insertion sites into the HBV genome using hydrodynamic tail vein injection into murine liver, demonstrating high levels of HBV surface antigen (HBsAg) and HBV DNA expression with rcccDNA formation. HBsAg expression from rcccDNA was >30,000 ng/mL over 78 days, while HBsAg-expression from pHBV1.3 plasmid DNA declined from 2,753 ng/mL to 131 ng/mL over that time in immunodeficient mice (P<0.001), reflective of plasmid DNA silencing. We then cloned Cre-recombinase in cis on the LoxP-HBV plasmids, achieving plasmid stability in bacteria with intron insertion into Cre and demonstrating rcccDNA formation after transfection in vitro and in vivo. These cis-Cre/LoxP-HBV plasmids were then used to create HBx-mutant and GFP reporter plasmids to further probe cccDNA biology and antiviral strategies against cccDNA. Overall, we believe these auto-generating rcccDNA plasmids will be of great value to model cccDNA for testing new therapies against HBV infection.

Genetic transformation of Rhodopseudomonas sphaeroides by plasmid DNA

1982 ◽  
Vol 152 (1) ◽  
pp. 89-97
Author(s):  
C S Fornari ◽  
S Kaplan
Keyword(s):  
Plasmid Dna ◽  
Recombinant Dna ◽  
Effective Means ◽  
Viable Cell ◽  
Broad Host Range ◽  
Dna Uptake ◽  
Rhodopseudomonas Sphaeroides ◽  
Open Circular ◽  
Polyethylene Glycol 6000

A broad-host-range cloning vector, pUI81, was constructed in vitro from plasmids RSF1010 and pSL25 (a pBR322 derivative) and used to assay for transformation in Rhodopseudomonas sphaeroides. Washing cells with 500 mM Tris was an effective means of inducing competence for DNA uptake. Transformation frequencies as high as 10(-5) (transformants per viable cell) have been achieved by incubating Tris-treated cells with plasmid DNA, 100 mM CaCl2, and 20% polyethylene glycol 6000. Maximum frequencies were obtained when recipient cells were spread onto selective media after a 6.5-h outgrowth period in antibiotic-free medium. The structure (open circular versus closed, covalent circular), size, and concentration of plasmid DNA all significantly affected the transformation frequency. Four different plasmids, all small and suitable as cloning vectors, have been introduced by transformation into several different R. sphaeroides strains. Recombinant DNA carried on small, nonconjugative plasmids with broad host ranges can now be directly transferred to R. sphaeroides by this method.

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