Gene engineered mesenchymal stem cells: greater transgene expression and efficacy with minicircle vs. plasmid DNA vectors in a mouse model of acute lung injury

Gene engineered mesenchymal stem cells: greater transgene expression and efficacy with minicircle vs. plasmid DNA vectors in a mouse model of acute lung injury

 Acute lung damage (ALI) and in its extreme kind, acute respiratory misery syndrome (ARDS), ends in elevated pulmonary vascular irritation and permeability and is a serious reason for mortality in lots of critically sick sufferers. Though cell-based therapies have proven promise in experimental ALI, methods are wanted to reinforce the efficiency of mesenchymal stem cells (MSCs) to develop more practical remedies. Genetic modification of MSCs has been demonstrated to considerably enhance the therapeutic advantages of those cells; nonetheless, the optimum vector for gene switch is just not clear. Given the acute nature of ARDS, transient transfection is fascinating to keep away from off-target results of long-term transgene expression, in addition to the potential antagonistic penalties of genomic integration.

 Right here, we explored whether or not a minicircle DNA (MC) vector containing human angiopoietin 1 (MC-ANGPT1) can present a more practical platform for gene-enhanced MSC remedy of ALI/ARDS. At 24 h after transfection, nuclear-targeted electroporation utilizing an MC-ANGPT1 vector resulted in a 3.7-fold better enhance in human ANGPT1 protein in MSC conditioned media in comparison with the usage of a plasmid ANGPT1  vector. Within the lipopolysaccharide (LPS)-induced ALI mannequin, administration of pANGPT1 transfected MSCs considerably decreased bronchoalveolar lavage (BAL) neutrophil counts by 57%, whereas MC-ANGPT1 transfected MSCs decreased it by 71% (p < 0.001) by Holm-Sidak’s a number of comparability take a look at.

Furthermore, in comparison with pANGPT1, the MC-ANGPT1 transfected MSCs considerably decreased pulmonary irritation, as noticed in decreased ranges of proinflammatory cytokines, akin to tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-2 (MIP-2). pANGPT1-transfected MSCs considerably decreased BAL albumin ranges by 71%, whereas MC-ANGPT1-transfected MSCs decreased it by 85%. Total, utilizing a minicircle vector, we demonstrated an environment friendly and sustained expression of the ANGPT1 transgene in MSCs and enhanced the therapeutic impact on the ALI mannequin in comparison with plasmid.

Plasmid Vectors for in Vivo Choice-Free Use with the Probiotic E. coli Nissle 1917

Escherichia coli Nissle 1917 (EcN) is a probiotic bacterium, generally employed to deal with sure gastrointestinal issues. It’s quick rising as an essential goal for the event of therapeutic engineered micro organism, benefiting from the wealth of information of E. coli biology and ease of manipulation. Bacterial artificial biology initiatives generally make the most of engineered plasmid vectors, that are easy to engineer and might reliably obtain excessive ranges of protein expression. Nevertheless, plasmids usually require antibiotics for upkeep, and the administration of an antibiotic is commonly incompatible with in vivo experimentation or remedy.

EcN natively accommodates plasmids pMUT1 and pMUT2, which don’t have any recognized operate however are secure inside the micro organism. Right here, we describe the event of the pMUT plasmids into a strong platform for engineering EcN for in vivo experimentation, alongside a CRISPR-Cas9 system to take away the native plasmids. We systematically engineered each pMUT plasmids to include choice markers, fluorescent markers, temperature delicate expression, and curli secretion programs to export a customizable purposeful materials into the extracellular area.

We then reveal that the engineered plasmids had been maintained in micro organism because the engineered micro organism go by the mouse GI tract with out choice, and that the secretion system stays purposeful, exporting functionalized curli proteins into the intestine. Our plasmid system presents a platform for the fast improvement of therapeutic EcN micro organism.

Gene engineered mesenchymal stem cells: greater transgene expression and efficacy with minicircle vs. plasmid DNA vectors in a mouse model of acute lung injury

Cloning in Plasmid Vectors: Blunt-Finish Cloning

This protocol describes procedures for cloning blunt-ended DNA fragments into linearized plasmid vectors. To acquire the utmost variety of “appropriate” ligation merchandise when cloning blunt-ended goal fragments, the 2 elements of DNA within the ligation response have to be current at an acceptable ratio. These outcomes assist the potential advantages of MC-ANGPT1 gene enhancement of MSC remedy to deal with ARDS.

If the molar ratio of plasmid vector to focus on DNA is just too excessive, then the ligation response might generate an undesirable variety of round empty plasmids, each monomeric and polymeric; if too low, the ligation response might generate an extra of linear and round homopolymers and heteropolymers of various sizes, orientations, and compositions. Because of this, the orientation of the overseas DNA and the variety of inserts in every recombinant clone should at all times be validated by restriction endonuclease mapping or another means.

Plasmid copy quantity variation of a modular vector set in Shewanella oneidensis MR-1

To advance artificial biology approaches that make the most of S. oneidensis as host for biotechnology functions, now we have investigated the variation in plasmid copy variety of a modular vector set ensuing from distinct origins of replication beneath totally different circumstances. The replicons yielded a ≈9X-fold vary for plasmid copy quantity variation in S. oneidensis (whereas the identical origins yielded a ≈3X-fold vary in Escherichia coli). This offers a sizeable vary to regulate gene expression ranges in S. oneidensis for artificial biology functions. As well as, plasmid harboring the pBBR1 origin resulted in secure copy numbers in S. oneidensis beneath totally different circumstances (mid-logarithmic, stationary, multi-plasmid).

Human Transcription factor p65 protein (NFKB3)

1-CSB-RP039874h
  • EUR 380.00
  • EUR 214.00
  • EUR 1309.00
  • EUR 560.00
  • EUR 873.00
  • EUR 262.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
Description: Recombinant Human Transcription factor p65 protein(NFKB3),partial expressed in E.coli

Human Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx570125-96tests 96 tests
EUR 668

Mouse Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx570127-96tests 96 tests
EUR 668

Sheep Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx364265-96tests 96 tests
EUR 926

Pig Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx361108-96tests 96 tests
EUR 825

Rabbit Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx362582-96tests 96 tests
EUR 825

Chicken Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx357192-96tests 96 tests
EUR 825

Monkey Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx359107-96tests 96 tests
EUR 825

Human Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx252815-96tests 96 tests
EUR 668

Mouse Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx254289-96tests 96 tests
EUR 668

Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit

20-abx156040
  • EUR 7237.00
  • EUR 3855.00
  • EUR 895.00
  • 10 × 96 tests
  • 5 × 96 tests
  • 96 tests

Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx255851-96tests 96 tests
EUR 668

Human Transcription factor p65 / NFKB3 (RELA) ELISA Kit

20-abx152945
  • EUR 6642.00
  • EUR 3542.00
  • EUR 825.00
  • 10 × 96 tests
  • 5 × 96 tests
  • 96 tests

Guinea pig Transcription factor p65 / NFKB3 (RELA) ELISA Kit

abx357262-96tests 96 tests
EUR 825

Anti-Anti-SEPT2 Antibody antibody

STJ25475 100 µl
EUR 277

Anti-Anti-SEPT5 Antibody antibody

STJ25477 100 µl
EUR 277
Description: This gene is a member of the septin gene family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. This gene is mapped to 22q11, the region frequently deleted in DiGeorge and velocardiofacial syndromes. A translocation involving the MLL gene and this gene has also been reported in patients with acute myeloid leukemia. Alternative splicing results in multiple transcript variants. The presence of a non-consensus polyA signal (AACAAT) in this gene also results in read-through transcription into the downstream neighboring gene (GP1BB; platelet glycoprotein Ib), whereby larger, non-coding transcripts are produced.

Anti-Anti-SEPT8 Antibody antibody

STJ25479 100 µl
EUR 277
Description: This gene is a member of the septin family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse, and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. Multiple alternatively spliced transcript variants encoding different isoforms have been found for this gene.

Anti-Anti-SEPT2 Antibody antibody

STJ28365 100 µl
EUR 277

Anti-Anti-SEPT7 Antibody antibody

STJ28963 100 µl
EUR 277
Description: This gene encodes a protein that is highly similar to the CDC10 protein of Saccharomyces cerevisiae. The protein also shares similarity with Diff 6 of Drosophila and with H5 of mouse. Each of these similar proteins, including the yeast CDC10, contains a GTP-binding motif. The yeast CDC10 protein is a structural component of the 10 nm filament which lies inside the cytoplasmic membrane and is essential for cytokinesis. This human protein functions in gliomagenesis and in the suppression of glioma cell growth, and it is required for the association of centromere-associated protein E with the kinetochore. Alternative splicing results in multiple transcript variants. Several related pseudogenes have been identified on chromosomes 5, 7, 9, 10, 11, 14, 17 and 19.

Anti-Anti-SEPT8 Antibody antibody

STJ117206 100 µl
EUR 277
Description: This gene is a member of the septin family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse, and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. Multiple alternatively spliced transcript variants encoding different isoforms have been found for this gene.

Anti-Anti-SEPT12 Antibody antibody

STJ117759 100 µl
EUR 277
Description: This gene encodes a guanine-nucleotide binding protein and member of the septin family of cytoskeletal GTPases. Septins play important roles in cytokinesis, exocytosis, embryonic development, and membrane dynamics. Multiple transcript variants encoding different isoforms have been found for this gene.

Anti-Anti-MARCH6 Antibody antibody

STJ118549 100 µl
EUR 277

Anti-Anti-MARCH6 Antibody antibody

STJ118550 100 µl
EUR 277

Anti-Anti-MARCH7 Antibody antibody

STJ118752 100 µl
EUR 277

Anti-Anti-SEPT3 Antibody antibody

STJ118990 100 µl
EUR 277

Anti-Anti-SEPT1 antibody antibody

STJ119580 100 µl
EUR 277
Description: This gene is a member of the septin family of GTPases. Members of this family are required for cytokinesis and the maintenance of cellular morphology. This gene encodes a protein that can form homo- and heterooligomeric filaments, and may contribute to the formation of neurofibrillary tangles in Alzheimer's disease. Alternatively spliced transcript variants have been found but the full-length nature of these variants has not been determined. [provided by RefSeq, Dec 2012]

Anti-Anti-SEPT7 Antibody antibody

STJ116214 100 µl
EUR 277
Description: This gene encodes a protein that is highly similar to the CDC10 protein of Saccharomyces cerevisiae. The protein also shares similarity with Diff 6 of Drosophila and with H5 of mouse. Each of these similar proteins, including the yeast CDC10, contains a GTP-binding motif. The yeast CDC10 protein is a structural component of the 10 nm filament which lies inside the cytoplasmic membrane and is essential for cytokinesis. This human protein functions in gliomagenesis and in the suppression of glioma cell growth, and it is required for the association of centromere-associated protein E with the kinetochore. Alternative splicing results in multiple transcript variants. Several related pseudogenes have been identified on chromosomes 5, 7, 9, 10, 11, 14, 17 and 19.

Anti-Anti-SEPT6 antibody antibody

STJ11100949 100 µl
EUR 277
Description: This gene is a member of the septin family of GTPases. Members of this family are required for cytokinesis. One version of pediatric acute myeloid leukemia is the result of a reciprocal translocation between chromosomes 11 and X, with the breakpoint associated with the genes encoding the mixed-lineage leukemia and septin 2 proteins. This gene encodes four transcript variants encoding three distinct isoforms. An additional transcript variant has been identified, but its biological validity has not been determined.

Anti-Anti-SEPT9 Antibody antibody

STJ111369 100 µl
EUR 277
Description: This gene is a member of the septin family involved in cytokinesis and cell cycle control. This gene is a candidate for the ovarian tumor suppressor gene. Mutations in this gene cause hereditary neuralgic amyotrophy, also known as neuritis with brachial predilection. A chromosomal translocation involving this gene on chromosome 17 and the MLL gene on chromosome 11 results in acute myelomonocytic leukemia. Multiple alternatively spliced transcript variants encoding different isoforms have been described.

Anti-Anti-SEPT11 Antibody antibody

STJ111530 100 µl
EUR 277

Anti-Anti-SEPT4 Antibody antibody

STJ112276 100 µl
EUR 277
Description: This gene is a member of the septin family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse, and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. This gene is highly expressed in brain and heart. Alternatively spliced transcript variants encoding different isoforms have been described for this gene. One of the isoforms (known as ARTS) is distinct; it is localized to the mitochondria, and has a role in apoptosis and cancer.

Anti-Anti-MARCH9 Antibody antibody

STJ112609 100 µl
EUR 277

Anti-Anti-SEPT11 Antibody antibody

STJ113941 100 µl
EUR 277

Anti-Anti-SEPT11 Antibody antibody

STJ114081 100 µl
EUR 277

Anti-Anti-SEPT5 Antibody antibody

STJ114819 100 µl
EUR 277
Description: This gene is a member of the septin gene family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. This gene is mapped to 22q11, the region frequently deleted in DiGeorge and velocardiofacial syndromes. A translocation involving the MLL gene and this gene has also been reported in patients with acute myeloid leukemia. Alternative splicing results in multiple transcript variants. The presence of a non-consensus polyA signal (AACAAT) in this gene also results in read-through transcription into the downstream neighboring gene (GP1BB; platelet glycoprotein Ib), whereby larger, non-coding transcripts are produced.

Anti-Anti-MARCH8 Antibody antibody

STJ114828 100 µl
EUR 277

Anti-Anti-DDB1 Antibody

A00333 100uL
EUR 455
Description: Rabbit Polyclonal DDB1 Antibody. Validated in IP and tested in Human, Mouse.

Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) ELISA Kit

AEA465Hu-10x96wellstestplate 10x96-wells test plate
EUR 5647.8
Description: This is Competitive Enzyme-linked immunosorbent assay for Antibody Detection.detection of Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) in serum, plasma and other biological fluids.

Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) ELISA Kit

AEA465Hu-1x48wellstestplate 1x48-wells test plate
EUR 552.76
Description: This is Competitive Enzyme-linked immunosorbent assay for Antibody Detection.detection of Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) in serum, plasma and other biological fluids.

Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) ELISA Kit

AEA465Hu-1x96wellstestplate 1x96-wells test plate
EUR 746.8
Description: This is Competitive Enzyme-linked immunosorbent assay for Antibody Detection.detection of Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) in serum, plasma and other biological fluids.

Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) ELISA Kit

AEA465Hu-5x96wellstestplate 5x96-wells test plate
EUR 3060.6
Description: This is Competitive Enzyme-linked immunosorbent assay for Antibody Detection.detection of Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) in serum, plasma and other biological fluids.

Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) ELISA Kit

4-AEA465Hu
  • EUR 5698.00
  • EUR 3011.00
  • EUR 747.00
  • 10 plates of 96 wells
  • 5 plates of 96 wells
  • 1 plate of 96 wells
Description: Enzyme-linked immunosorbent assay based on the Competitive Inhibition method for detection of Human Anti-Anti-Sperm Antibody Antibody (Anti-AsAb) in samples from serum, plasma and other biological fluids with no significant corss-reactivity with analogues from other species.

ELISA kit for Human Anti-AsAb (Anti-Anti-Sperm Antibody Antibody)

ELK8071 1 plate of 96 wells
EUR 432
Description: A competitive Inhibition ELISA kit for detection of Anti-Anti-Sperm Antibody Antibody from Human in samples from blood, serum, plasma, cell culture fluid and other biological fluids.

Anti- ZNF660 antibody

ABD2410 100 ug
EUR 438

Anti-STAT5B Antibody

A1753-100
EUR 479

Anti-CREB1 Antibody

A1754-100
EUR 479

Anti-COX7B Antibody

A1756-100
EUR 479

Anti-COX6B1 Antibody

A1757-100
EUR 479

Anti-COX7B2 Antibody

A1758-100
EUR 479

Anti-COX19 Antibody

A1759-100
EUR 479

Anti-AIMP2 Antibody

A1760-100
EUR 479

Anti-NFKBIE Antibody

A1762-100
EUR 479

Anti-NFKBID Antibody

A1764-100
EUR 479

Anti-APIP Antibody

A1766-100
EUR 479

Anti-BAG1 Antibody

A1768-100
EUR 479

Anti-IL10RA Antibody

A1770-100
EUR 479

Anti-IL10RB Antibody

A1771-100
EUR 479

Anti-IL12RB1 Antibody

A1772-100
EUR 479

Anti-IL13RA1 Antibody

A1773-100
EUR 479

Anti-IL13RA2 Antibody

A1774-100
EUR 479

Anti-IL17RC Antibody

A1775-100
EUR 479

Anti-IL18R1 Antibody

A1776-100
EUR 479

Anti-IL1R1 Antibody

A1777-100
EUR 479

Anti- TauD Antibody

A1804-100
EUR 370

Anti- TauD Antibody

A1804-30T
EUR 146

Anti-KRT20 Antibody

A1859-100
EUR 403

Anti-PGR Antibody

A1860-100
EUR 403

Anti-HSPA9 Antibody

A1861-100
EUR 403

Anti-ERBB4 Antibody

A1862-100
EUR 403

Anti-CA12 Antibody

A1865-100
EUR 403

Anti-TPD52 Antibody

A1867-100
EUR 403

Anti-STRADA Antibody

A1868-100
EUR 403

Anti-CD276 Antibody

A1869-100
EUR 403

Anti-ICT1 Antibody

A1872-100
EUR 403

Anti-OCIAD1 Antibody

A1873-100
EUR 403

Anti-BRINP1 Antibody

A1874-100
EUR 403

Anti-WT1 Antibody

A1875-100
EUR 403

Anti-SART3 Antibody

A1879-100
EUR 403

Anti-RRBP1 Antibody

A1880-100
EUR 403

Anti-KIAA1524 Antibody

A1881-100
EUR 403

Anti-ENO2 Antibody

A1883-100
EUR 403

Anti-GPC3 Antibody

A1884-100
EUR 403

Anti-ACP1 Antibody

A1885-100
EUR 403

Anti-Elafin Antibody

A1887-100
EUR 403

Anti-AVEN Antibody

A1891-100
EUR 403

Anti-MST1 Antibody

A1895-100
EUR 403

Anti-WSB2 Antibody

A1896-100
EUR 403

Anti-UPP1 Antibody

A1898-100
EUR 403

Anti-YY1AP1 Antibody

A1899-100
EUR 403

Anti-CDK5RAP3 Antibody

A1907-100
EUR 403

Anti-TCERG1 Antibody

A1909-100
EUR 403

Anti-CD44 Antibody

A1910-100
EUR 403

Anti-TIMM17A Antibody

A1911-100
EUR 403

Anti-CYP2W1 Antibody

A1912-100
EUR 403

Anti-NDUFAF2 Antibody

A1913-100
EUR 403

Anti-PRAME Antibody

A1914-100
EUR 403

Anti-S100P Antibody

A1915-100
EUR 403

Anti-HES3 Antibody

A1636-100
EUR 479

Anti-HES5 Antibody

A1637-100
EUR 479

Anti-HMGA2 Antibody

A1638-100
EUR 479

Anti-HuD Antibody

A1639-100
EUR 370

Anti-ITGA4 Antibody

A1640-100
EUR 479

Anti-IPMK Antibody

A1641-100
EUR 479

Anti-LHX2 Antibody

A1643-100
EUR 479

Anti-MAP2 Antibody

A1644-100
EUR 479

Anti-MSI1 Antibody

A1646-100
EUR 479

Anti-NEDD1 Antibody

A1647-100
EUR 370

Anti-NEUROD2 Antibody

A1648-100
EUR 370

Anti-NGFR Antibody

A1650-100
EUR 479

Anti-OLFM1 Antibody

A1651-100
EUR 370

Anti-NR2E1 Antibody

A1654-100
EUR 479

Anti-OLIG2 Antibody

A1656-100
EUR 479

Anti-PAX3 Antibody

A1657-100
EUR 370

Anti-PLAGL1 Antibody

A1658-100
EUR 479

Anti- S100B Antibody

A1659-100
EUR 479

Anti-SLUG Antibody

A1660-100
EUR 370

Anti-SOX1 Antibody

A1661-100
EUR 479

Anti-SOX11 Antibody

A1662-100
EUR 370

Anti-EOMES Antibody

A1665-100
EUR 370

Anti-TGIF1 Antibody

A1667-100
EUR 479

Anti-CD9 Antibody

A1668-100
EUR 370

Anti-CD168 Antibody

A1671-100
EUR 479

This will allow the belief of artificial circuits in S. oneidensis the place predictable, quantitative conduct is desired (in both single- or double-plasmid contexts). Klebsiella pneumoniae is a bacterial pathogen with rising charges of resistance to carbapenem antibiotics, however the inhabitants construction and genetic drivers of carbapenem-resistant Okay pneumoniae (CRKP) stay underexplored in creating nations. Carbapenem-resistant Okay pneumoniae had been not too long ago launched into Peru however have grown quickly in prevalence, enabling research of this pathogen because it expands into an unaffected setting.

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