cationic lipid  (Thermo Fisher)


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    Name:
    Lipofectamine Transfection Reagent
    Description:
    Lipofectamine Transfection Reagent is one of our first generation cationic lipid transfection reagents formulated for the transfection of DNA into eukaryotic cells Lipofectamine Transfection Reagent is a trusted value reagent with many years of cited use in thousands of publications and with many cell lines With Lipofectamine Transfection Reagent you ll get • A reagent proven to work in high throughput applications• A reliable reagent for establishing stable cell lines• A reagent that works well with PLUS Reagent for higher protein expressionA proven technologyOur Lipofectamine brand reagents have been recognized as the most cited transfection reagent family with tens of thousands of citations since launch in 1993 Lipofectamine Transfection Reagent is no exception as a trusted reagent for a wide range of cell lines When used in combination with PLUS Reagent Lipofectamine Transfection Reagent has been shown to work well with cells such as BHK 21 NIH 3T3 COS 1 fibroblasts keratinocytes HT 29 MRC 5 and SK BR3 A value alternative for plasmid DNAWith such a legacy comes a commitment to providing the best reagents for all scientists and labs regardless of their funding capabilities Lipofectamine Transfection Reagent is a value alternative for transfection of plasmid DNA into eukaryotic cells with a simplified protocol For optimal results in a wider range of plasmid DNA transfection conditions we recommend Lipofectamine LTX Reagent due to its optimal balance of potency low cytotoxicity Lipofectamine 2000 Transfection Reagent is recommended if you are transfecting a broad range of cell lines or want to transfect RNA
    Catalog Number:
    18324010
    Price:
    None
    Category:
    Cell Culture Transfection Reagents
    Applications:
    Cell Culture|Plasmid Transfection|Stem Cell & Primary Cell Transfections|Transfection
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    Structured Review

    Thermo Fisher cationic lipid
    Lipofectamine Transfection Reagent is one of our first generation cationic lipid transfection reagents formulated for the transfection of DNA into eukaryotic cells Lipofectamine Transfection Reagent is a trusted value reagent with many years of cited use in thousands of publications and with many cell lines With Lipofectamine Transfection Reagent you ll get • A reagent proven to work in high throughput applications• A reliable reagent for establishing stable cell lines• A reagent that works well with PLUS Reagent for higher protein expressionA proven technologyOur Lipofectamine brand reagents have been recognized as the most cited transfection reagent family with tens of thousands of citations since launch in 1993 Lipofectamine Transfection Reagent is no exception as a trusted reagent for a wide range of cell lines When used in combination with PLUS Reagent Lipofectamine Transfection Reagent has been shown to work well with cells such as BHK 21 NIH 3T3 COS 1 fibroblasts keratinocytes HT 29 MRC 5 and SK BR3 A value alternative for plasmid DNAWith such a legacy comes a commitment to providing the best reagents for all scientists and labs regardless of their funding capabilities Lipofectamine Transfection Reagent is a value alternative for transfection of plasmid DNA into eukaryotic cells with a simplified protocol For optimal results in a wider range of plasmid DNA transfection conditions we recommend Lipofectamine LTX Reagent due to its optimal balance of potency low cytotoxicity Lipofectamine 2000 Transfection Reagent is recommended if you are transfecting a broad range of cell lines or want to transfect RNA
    https://www.bioz.com/result/cationic lipid/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cationic lipid - by Bioz Stars, 2021-03
    86/100 stars

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    Transfection:

    Article Title: Effect of silencing lncRNATUG1 on rapamycin-induced inhibition of endothelial cell proliferation and migration
    Article Snippet: siRNA specifically targeting TUG1 (siTUG1: siTUG1-1, 5′-GCUUGGCUUCUAUUCUGAAUCCUUU-3′; siTUG1-2, 5′-CAGCUGUUACCAUUCAACUUCUUAA-3′) and negative control siRNA (siNC, 5′-UUCUCCGAACGUGUCACGUTT-3′) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China). .. Once HUVECs reached 50–70% confluence, they were seeded in 6-well plates and transfected with 100 nM siTUG1 and siNC using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. .. At 24 h post-transfection, DMEM and 100 ng/ml rapamycin (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) were added.

    Article Title: MiR-1180-5p regulates apoptosis of Wilms’ tumor by targeting p73
    Article Snippet: MiR-1180 inhibitor and scramble, which were labeled with green fluorescence protein (GFP), were purchased from Genepharma (Suzhou, China). .. Cell transfection was performed using Lipofectamine RNAiMax (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. .. A colorimetric assay with tetrazolium salt was used to assess the antiproliferative effects.

    Article Title: Immunogene therapy with fusogenic nanoparticles modulates macrophage response to Staphylococcus aureus
    Article Snippet: For lipids, DMPC, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(PEG)-2000], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(PEG)-2000], and DOTAP were purchased from Avanti Polar Lipids (Alabaster, AL) and stored at − 4 °C. .. Fluorescent dyes Calcein (Sigma-Aldrich) and hydrophobic DiI (Life Technologies) were used and Lipofectamine® 2000 Transfection Reagent was obtained from Thermo Fisher Scientific. .. Custom siRNAs were purchased from Dharmacon (Lafayette, CO) and primers were purchased from IDT DNA (San Diego, CA).

    Article Title: Transformation of human liver L-O2 cells mediated by stable HBx transfection
    Article Snippet: To replicate the role of HBx gene in another cell line, we attempted to establish a stably HBx-transfected NIH/3T3 cell line. .. NIH/3T3 cells were transfected with Lipofectamine (Invitrogen, USA) as described previously . .. The plasmids pcDNA3 and pCMV-X were used in the transfection experiments.

    Article Title: Inhibition of mTORC2/RICTOR Impairs Melanoma Hepatic Metastasis
    Article Snippet: Si); Invitrogen, Waltham, MA, USA] using Lipofectamine RNAiMAX transfection reagent (Invitrogen, Waltham, MA, USA). .. Briefly, pre-plated cells were incubated with transfection mixture [Opti-MEM® (Gibco, Waltham, MA, USA), Lipofectamine 2.5% vol/vol, siRNA 50 nM] for 6 hours. .. 48 hours after transfection knock-down efficiency was confirmed by Western blotting and tumor cells were processed for further experiments.

    Article Title: The p300/YY1/miR-500a-5p/HDAC2 signalling axis regulates cell proliferation in human colorectal cancer
    Article Snippet: .. Cell transfection was performed with Lipofectamine TM 2000 (Invitrogen) as described in the manufacturer’s protocol. .. Each group of isolated tumour cells was seeded onto 96-well plates in triplicate at a density of 103 /well and incubated for 48 h. Subsequently, the cells were incubated for an additional 2 h in the respective media containing 50 μM EdU (RiboBio, Guangzhou, China).

    Article Title: Aurora Kinase A proximity interactome reveals centriolar satellites as regulators of its function during primary cilium biogenesis
    Article Snippet: All cell lines were tested for mycoplasma by MycoAlert Mycoplasma Detection Kit (Lonza). .. RPE1 cells were transfected with the plasmids using Lipofectamine Lipofectamine for DNA transfections and Lipofectamine RNAiMax for siRNA transfections according to the manufacturer’s instructions (Thermo Fisher Scientific). ..

    Article Title: MicroRNA-17-5p aggravates lipopolysaccharide-induced injury in nasal epithelial cells by targeting Smad7
    Article Snippet: Scramble, siNC, si-miR-17-5p, and miR-17-5p mimic were synthesized by GenePharma Co (Shanghai, China). .. Cell transfections were conducted using Lipofectamine 3000 reagent (Invitrogen) as per the manufacturer’s protocol. .. RNAs from the cultured cells were extracted using RNA pure Rapid Extraction Kit (Bioteke Corporation, Beijing, China) according to the manufacturer’s instructions.

    Incubation:

    Article Title: Inhibition of mTORC2/RICTOR Impairs Melanoma Hepatic Metastasis
    Article Snippet: Si); Invitrogen, Waltham, MA, USA] using Lipofectamine RNAiMAX transfection reagent (Invitrogen, Waltham, MA, USA). .. Briefly, pre-plated cells were incubated with transfection mixture [Opti-MEM® (Gibco, Waltham, MA, USA), Lipofectamine 2.5% vol/vol, siRNA 50 nM] for 6 hours. .. 48 hours after transfection knock-down efficiency was confirmed by Western blotting and tumor cells were processed for further experiments.

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    Thermo Fisher cationic lipid based transfection system
    FKCU/LTR reporter cells, transfected with the pHook-3/tas plasmid and stained by the β -galassay. <t>Transfection</t> efficiency approximated as 25–30%, based on the development of blue-coloured cells. (Magnification ×200).
    Cationic Lipid Based Transfection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cationic lipid based transfection system/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cationic lipid based transfection system - by Bioz Stars, 2021-03
    97/100 stars
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    FKCU/LTR reporter cells, transfected with the pHook-3/tas plasmid and stained by the β -galassay. Transfection efficiency approximated as 25–30%, based on the development of blue-coloured cells. (Magnification ×200).

    Journal: Journal of Feline Medicine and Surgery

    Article Title: FIP: a novel approach to vaccination

    doi: 10.1016/j.jfms.2003.08.010

    Figure Lengend Snippet: FKCU/LTR reporter cells, transfected with the pHook-3/tas plasmid and stained by the β -galassay. Transfection efficiency approximated as 25–30%, based on the development of blue-coloured cells. (Magnification ×200).

    Article Snippet: A cationic lipid-based transfection system (Lipofectamine, Invitrogen) was used to carry plasmid DNA into FKCU/LTR/zeo/β -gal reporter cells (a kind gift from Prof. A. Rethwilm, University of Wurzburg, Germany).

    Techniques: Transfection, Plasmid Preparation, Staining

    Viral dsRNA complexed with cationic lipids activates mesangial cells to produce high amounts of Il-6 and type I IFN via a Trif-independent pathway. Primary mesangial cells were isolated from wild-type C56BL/6 mice and Trif -mutant mice as described in Materials and Methods . Cells were stimulated with increasing doses of poly I:C (pI:C) RNA alone or complexed with the cationic lipid (CL) Lipofectamine. Supernatants were harvested after 24 hours and analyzed by ELISA for the following mediators: Il-6 ( A ), IFN-α ( B ), and IFN-β ( C ). Data are means ± SEM from three experiments each analyzed in duplicate and presented in a logarithmic scale. ∗ P

    Journal: The American Journal of Pathology

    Article Title: Viral RNA Induces Type I Interferon-Dependent Cytokine Release and Cell Death in Mesangial Cells via Melanoma-Differentiation-Associated Gene-5

    doi: 10.2353/ajpath.2009.080585

    Figure Lengend Snippet: Viral dsRNA complexed with cationic lipids activates mesangial cells to produce high amounts of Il-6 and type I IFN via a Trif-independent pathway. Primary mesangial cells were isolated from wild-type C56BL/6 mice and Trif -mutant mice as described in Materials and Methods . Cells were stimulated with increasing doses of poly I:C (pI:C) RNA alone or complexed with the cationic lipid (CL) Lipofectamine. Supernatants were harvested after 24 hours and analyzed by ELISA for the following mediators: Il-6 ( A ), IFN-α ( B ), and IFN-β ( C ). Data are means ± SEM from three experiments each analyzed in duplicate and presented in a logarithmic scale. ∗ P

    Article Snippet: Primary mesangial cells were stimulated with endotoxin-free poly I:C RNA (InvivoGen, Toulouse, France), poly I:C RNA transfected with the cationic lipid Lipofectamine 2000 (Invitrogen, Carlsbad, CA) or ultrapure LPS (InvivoGen) for 24 hours in RPMI 1640 containing 5% FCS in the presence or absence of murine IFN-α (AbD Serotec, Oxford, UK), IFN-β (PBL, Piscataway, NJ), or IFN-γ (PeproTech, Rocky Hill, NJ).

    Techniques: Isolation, Mouse Assay, Mutagenesis, Enzyme-linked Immunosorbent Assay

    Transgene integration frequencies of linear covalently closed (LCC) versus conventional circular covalently closed (CCC) pDNA into the human genome . We directed site-specific insertion of LCC or CCC DNA constructs into the human (HEK 293) genome using the commercially available Flp-In System (Invitrogen) that exploits the Flp- FRT- mediated recombination system. Efficacy of LCC and standard pDNA vector chromosomal integration were compared by Flp- FTR -mediated site-specific integration to verify and compare the safety profile of LCC versus standard pDNA vectors. Flp-In 293 HEK cells (Invitrogen) carrying the attachment site ( att ), were transfected by the Flp-In kit plasmids pcDNA5/FRT and pcDNA5/FRT/CAT that possess the Flp target site FRT and the indicator chloramphenicol acetyl tranferase ( CAT ). The super sequence was cloned into each plasmid and passed through Tel + R-cells to generate plasmids with LCC topology. Transfection was performed in the presence or absence of Flp-recombinase expression vector, pOG44, for site-specific single crossover or random integration, respectively. Transfected cells were selected with hygromycin for 3 weeks, and resistant colonies were counted as a measure of colony forming units. Mean ± SEM values are shown ( n = 3–5). The Hyg R , Zeo S , and B-Gal − cells represent the result of Flp-mediated site-specific insertion of the gene of interest. In contrast, Hyg R , Zeo R , and B-Gal + cells represent low-level illegitimate integration events. The mean was calculated from a minimum of three trials and with different plasmids. Integration frequency (IF) is expressed as the fraction of integrant cells to total transfected cells. IF of LCC was significantly lower than its CCC counterpart and control plasmid with no super sequence, ( P

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: DNA Ministrings: Highly Safe and Effective Gene Delivery Vectors

    doi: 10.1038/mtna.2014.16

    Figure Lengend Snippet: Transgene integration frequencies of linear covalently closed (LCC) versus conventional circular covalently closed (CCC) pDNA into the human genome . We directed site-specific insertion of LCC or CCC DNA constructs into the human (HEK 293) genome using the commercially available Flp-In System (Invitrogen) that exploits the Flp- FRT- mediated recombination system. Efficacy of LCC and standard pDNA vector chromosomal integration were compared by Flp- FTR -mediated site-specific integration to verify and compare the safety profile of LCC versus standard pDNA vectors. Flp-In 293 HEK cells (Invitrogen) carrying the attachment site ( att ), were transfected by the Flp-In kit plasmids pcDNA5/FRT and pcDNA5/FRT/CAT that possess the Flp target site FRT and the indicator chloramphenicol acetyl tranferase ( CAT ). The super sequence was cloned into each plasmid and passed through Tel + R-cells to generate plasmids with LCC topology. Transfection was performed in the presence or absence of Flp-recombinase expression vector, pOG44, for site-specific single crossover or random integration, respectively. Transfected cells were selected with hygromycin for 3 weeks, and resistant colonies were counted as a measure of colony forming units. Mean ± SEM values are shown ( n = 3–5). The Hyg R , Zeo S , and B-Gal − cells represent the result of Flp-mediated site-specific insertion of the gene of interest. In contrast, Hyg R , Zeo R , and B-Gal + cells represent low-level illegitimate integration events. The mean was calculated from a minimum of three trials and with different plasmids. Integration frequency (IF) is expressed as the fraction of integrant cells to total transfected cells. IF of LCC was significantly lower than its CCC counterpart and control plasmid with no super sequence, ( P

    Article Snippet: Cationic polymer transfection reagents (jetPRIME) were obtained from VWR and cationic lipid transfection reagents (Lipofectamine 2000, Lipofectamine LTX, and Plus reagents) from Invitrogen.

    Techniques: Countercurrent Chromatography, Construct, Plasmid Preparation, Transfection, Sequencing, Clone Assay, Expressing

    Enhanced minivectors confer superior transfection efficiency in epithelial and cancer cells . Enhanced pNN9 was processed into minicircle and ministring vectors by passing through Cre + and Tel/TelN + R-cells, respectively. DNA vectors were mixed by cationic polymer and cationic lipid carriers and transfected into cancer and epithelial cells. Cells were collected 48 hours post-transfection and analyzed by fluorescence-activated cell sorting for green fluorescent protein expression and cytotoxicity of cationic lipid/polymer carriers. Parent pNN9 is a 5.6 kb DNA molecule carrying two super sequence sites flanking the minimal eGFP cistron. Unlike pNN9, pNN7 does not possess SV40E, limiting its nuclear translocation efficiency particularly in nondividing cells. Transfection efficiency (TE) was measured as the number of eGFP-expressing cells divided by the total number of cells. Propidium iodide was added to measure synthetic carrier toxicity. ( a ) Slowly dividing epithelial cells and ( b ) rapidly dividing cancer cells transfected by 5 μg polyplexed or lipoplexed DNA vector. In both cell lines, mini DNA vectors show significantly higher TE ( P ≤ 0.001). In epithelial HEK cells, ministring DNA showed significantly higher TE versus its minicircle counterpart ( P

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: DNA Ministrings: Highly Safe and Effective Gene Delivery Vectors

    doi: 10.1038/mtna.2014.16

    Figure Lengend Snippet: Enhanced minivectors confer superior transfection efficiency in epithelial and cancer cells . Enhanced pNN9 was processed into minicircle and ministring vectors by passing through Cre + and Tel/TelN + R-cells, respectively. DNA vectors were mixed by cationic polymer and cationic lipid carriers and transfected into cancer and epithelial cells. Cells were collected 48 hours post-transfection and analyzed by fluorescence-activated cell sorting for green fluorescent protein expression and cytotoxicity of cationic lipid/polymer carriers. Parent pNN9 is a 5.6 kb DNA molecule carrying two super sequence sites flanking the minimal eGFP cistron. Unlike pNN9, pNN7 does not possess SV40E, limiting its nuclear translocation efficiency particularly in nondividing cells. Transfection efficiency (TE) was measured as the number of eGFP-expressing cells divided by the total number of cells. Propidium iodide was added to measure synthetic carrier toxicity. ( a ) Slowly dividing epithelial cells and ( b ) rapidly dividing cancer cells transfected by 5 μg polyplexed or lipoplexed DNA vector. In both cell lines, mini DNA vectors show significantly higher TE ( P ≤ 0.001). In epithelial HEK cells, ministring DNA showed significantly higher TE versus its minicircle counterpart ( P

    Article Snippet: Cationic polymer transfection reagents (jetPRIME) were obtained from VWR and cationic lipid transfection reagents (Lipofectamine 2000, Lipofectamine LTX, and Plus reagents) from Invitrogen.

    Techniques: Transfection, Fluorescence, FACS, Expressing, Sequencing, Translocation Assay, Plasmid Preparation

    Effect of pDNA size and topology on intracellular kinetics . Confocal microscopic analysis of the uptake and distribution (panel a ) and eGFP expression (panel b ) of minicircle and ministring DNA in HEK 293 cells. Cells were imaged at 3 and 7 hours after dosing with the three different Cy5-labelled vectors complexed with Lipofectamine at 37 °C under two separate staining conditions: the first row of panel a 3- and 7-hour sections shows cells stained with SYTO 21 green nuclear stain and CellMask Orange cell membrane stain and the second row shows cells stained with LysoTracker green and CellMask Orange. The four pictures in the second rows represent signals in the three separate channels for LysoTracker green, CellMask orange, Cy5 and all combined (clockwise). Panel b shows eGFP expression for the three different vectors at 8 and 12 hours after incubation after removing the transfection complexes (transfection time was 6 hours).

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: DNA Ministrings: Highly Safe and Effective Gene Delivery Vectors

    doi: 10.1038/mtna.2014.16

    Figure Lengend Snippet: Effect of pDNA size and topology on intracellular kinetics . Confocal microscopic analysis of the uptake and distribution (panel a ) and eGFP expression (panel b ) of minicircle and ministring DNA in HEK 293 cells. Cells were imaged at 3 and 7 hours after dosing with the three different Cy5-labelled vectors complexed with Lipofectamine at 37 °C under two separate staining conditions: the first row of panel a 3- and 7-hour sections shows cells stained with SYTO 21 green nuclear stain and CellMask Orange cell membrane stain and the second row shows cells stained with LysoTracker green and CellMask Orange. The four pictures in the second rows represent signals in the three separate channels for LysoTracker green, CellMask orange, Cy5 and all combined (clockwise). Panel b shows eGFP expression for the three different vectors at 8 and 12 hours after incubation after removing the transfection complexes (transfection time was 6 hours).

    Article Snippet: Cationic polymer transfection reagents (jetPRIME) were obtained from VWR and cationic lipid transfection reagents (Lipofectamine 2000, Lipofectamine LTX, and Plus reagents) from Invitrogen.

    Techniques: Expressing, Staining, Incubation, Transfection

    Effect of SV40 enhancer sequence on transfection efficiency . Lipoplexed/polyplexed DNA vectors were transfected into cancer and epithelial cells. Cells were collected 48 hours post-transfection and analyzed by FACS. Transfection efficiency (TE) was determined as the number of GFP-expressing cells divided by the total number of cells. Propidium iodide was added to measure transfection reagent cytotoxicity. In both HEK 293 and OVCAR-3 cells, ( a ) lipoplexed and ( b ) polyplexed pNN9 vectors (with 4 SV40E) show significantly higher TE compared with their pNN7 (with no SV40E) counterparts (P ≤ 0.001). DTS, DNA targeting sequences. FACS, fluorescence-activated cell sorting; GFP, green fluorescent protein.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: DNA Ministrings: Highly Safe and Effective Gene Delivery Vectors

    doi: 10.1038/mtna.2014.16

    Figure Lengend Snippet: Effect of SV40 enhancer sequence on transfection efficiency . Lipoplexed/polyplexed DNA vectors were transfected into cancer and epithelial cells. Cells were collected 48 hours post-transfection and analyzed by FACS. Transfection efficiency (TE) was determined as the number of GFP-expressing cells divided by the total number of cells. Propidium iodide was added to measure transfection reagent cytotoxicity. In both HEK 293 and OVCAR-3 cells, ( a ) lipoplexed and ( b ) polyplexed pNN9 vectors (with 4 SV40E) show significantly higher TE compared with their pNN7 (with no SV40E) counterparts (P ≤ 0.001). DTS, DNA targeting sequences. FACS, fluorescence-activated cell sorting; GFP, green fluorescent protein.

    Article Snippet: Cationic polymer transfection reagents (jetPRIME) were obtained from VWR and cationic lipid transfection reagents (Lipofectamine 2000, Lipofectamine LTX, and Plus reagents) from Invitrogen.

    Techniques: Sequencing, Transfection, FACS, Expressing, Fluorescence

    Linear covalently closed (LCC) single crossover integration into human cell induces apoptotic cell death . Any surviving LCC and circular covalently closed (CCC) pDNA-mediated Hyg R , Zeo S , B-Gal − cells were isolated and expanded in six-well culture plates. To determine fate of these cells after site-specific insertion of the LCC and standard pDNA vectors, expanded clones were collected and pooled at 3, 5, and 7 weeks post-transfection. To determine whether cells that had integrated the LCC vector now possessed, as hypothesized in Figure 5b, a chromosomal disruption at the site of vector integration, 106 cells of each group were stained by Annexin V-FITC, and propidium iodide (PI) and results were analyzed by flow cytometry. Measurements were normalized against controls: (i) unstained nonintegrated cells, (ii) nonintegrated cells stained with only Annexin V, and (iii) nonintegrated cells stained with only PI. ( a ) Two-channel reading of flow cytometry results aligned with cell morphology of CCC (upper) and LCC (bottom) integrants. ( b ) Morphology of integrated cells. Graphs represent percentage of healthy, apoptotic, or necrotic cells. Mean of a minimum of three trials. LCC integrants show significantly lower viability and higher apoptotic and necrotic index compared with the CCC integrants ( P

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: DNA Ministrings: Highly Safe and Effective Gene Delivery Vectors

    doi: 10.1038/mtna.2014.16

    Figure Lengend Snippet: Linear covalently closed (LCC) single crossover integration into human cell induces apoptotic cell death . Any surviving LCC and circular covalently closed (CCC) pDNA-mediated Hyg R , Zeo S , B-Gal − cells were isolated and expanded in six-well culture plates. To determine fate of these cells after site-specific insertion of the LCC and standard pDNA vectors, expanded clones were collected and pooled at 3, 5, and 7 weeks post-transfection. To determine whether cells that had integrated the LCC vector now possessed, as hypothesized in Figure 5b, a chromosomal disruption at the site of vector integration, 106 cells of each group were stained by Annexin V-FITC, and propidium iodide (PI) and results were analyzed by flow cytometry. Measurements were normalized against controls: (i) unstained nonintegrated cells, (ii) nonintegrated cells stained with only Annexin V, and (iii) nonintegrated cells stained with only PI. ( a ) Two-channel reading of flow cytometry results aligned with cell morphology of CCC (upper) and LCC (bottom) integrants. ( b ) Morphology of integrated cells. Graphs represent percentage of healthy, apoptotic, or necrotic cells. Mean of a minimum of three trials. LCC integrants show significantly lower viability and higher apoptotic and necrotic index compared with the CCC integrants ( P

    Article Snippet: Cationic polymer transfection reagents (jetPRIME) were obtained from VWR and cationic lipid transfection reagents (Lipofectamine 2000, Lipofectamine LTX, and Plus reagents) from Invitrogen.

    Techniques: Countercurrent Chromatography, Isolation, Clone Assay, Transfection, Plasmid Preparation, Staining, Flow Cytometry, Cytometry

    SPC25 knockdown inhibits the proliferation of PC-3 cells. (A) Expression of SPC25 mRNA following transfection with the indicated shRNAs in PC-3 cells. (B) An MTT assay demonstrated that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (C) The Celigo ® system revealed that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (D) Cell number of shCtrl or shSPC25 group in each day was calculated using the Celigo ® system. P

    Journal: Oncology Letters

    Article Title: Knockdown of spindle pole body component 25 homolog inhibits cell proliferation and cycle progression in prostate cancer

    doi: 10.3892/ol.2018.8003

    Figure Lengend Snippet: SPC25 knockdown inhibits the proliferation of PC-3 cells. (A) Expression of SPC25 mRNA following transfection with the indicated shRNAs in PC-3 cells. (B) An MTT assay demonstrated that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (C) The Celigo ® system revealed that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (D) Cell number of shCtrl or shSPC25 group in each day was calculated using the Celigo ® system. P

    Article Snippet: Opti-modified Eagle's medium (Opti-MEM) which was ideal for use during cationic lipid transfections especially Lipofectamine 2000 transfection reagents was purchased from Thermo Fisher Scientific, Inc (cat. no. 31985062).

    Techniques: Expressing, Transfection, MTT Assay