• Plasmid transfection of mammalian cells is the dominant platform used to produce adeno-associated virus (AAV) vectors for clinical and research applications. (cytovax.com)
  • Therapeutic Application of Extracellular Vesicles-Capsulated Adeno-Associated Virus Vector via nSMase2/Smpd3 , Satellite, and Immune Cells in Duchenne Muscular Dystrophy. (cytovax.com)
  • Our proficiency with adeno-associated virus (AAV), lentivirus, adenovirus, and other vectors helps streamline the development and manufacturing of cell and gene therapies. (sigmaaldrich.com)
  • Adeno-associated virus (AAV) and Lentivirus (LV) are the most common viral vectors used in gene therapy bioprocessing. (repligen.com)
  • Several virus types have been studied for use as viral vectors in gene therapy, including adenovirus, lentivirus, retrovirus, and adeno-associated virus (AAV). (biopharminternational.com)
  • The Effectene Transfection Reagent is intended for molecular biology applications. (qiagen.com)
  • Effectene Transfection Reagent is a nonliposomal lipid reagent for DNA transfection into a broad range of cell types. (qiagen.com)
  • Due to its low cytotoxicity, Effectene Transfection Reagent is highly suitable for transfection of primary cells and many sensitive cell lines. (qiagen.com)
  • Transfection using Effectene Transfection Reagent requires low amounts of DNA and minimal handling. (qiagen.com)
  • In addition, removal of transfection complexes is not required, making this reagent highly suitable for high-throughput screening. (qiagen.com)
  • Lipofectamine 3000 Transfection Reagent delivers a high efficiency for hard-to-transfect cell lines. (thermofisher.com)
  • When performing transfections in hard-to-transfect cell lines, Lipofectamine 3000 reagent can be an ideal choice. (thermofisher.com)
  • Lipofectamine 3000 reagent leverages our most advanced lipid nanoparticle technology to enable exceptional transfection performance and reproducible results. (thermofisher.com)
  • Each reagent was used to transfect HEK 293, HeLa, LNCaP, HepG2, and A549 cell lines in a 96-well format, and GFP expression was analyzed 48 hours post-transfection. (thermofisher.com)
  • Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines. (thermofisher.com)
  • This superior transfection performance allows you to reduce the reagent dose, thus helping lower toxicity risk when transfecting your cell line. (thermofisher.com)
  • Each reagent was used to transfect HeLa cells in a 96-well format at the indicated doses with an emerald green fluorescent protein (GFP)-expressing vector. (thermofisher.com)
  • Lipofectamine 3000 reagent is a specially formulated gentle transfection reagent that is optimized for plasmid delivery into CHO cells with high efficiency and low cell toxicity. (thermofisher.com)
  • The reagent allows for transfection to be performed both in presence or absence of serum. (thermofisher.com)
  • The easy protocol, exceptional plasmid DNA delivery, and the ability to effectively generate stable cell lines makes Lipofectamine 3000 reagent, the perfect reagent for CHO cell transfection experiments. (thermofisher.com)
  • We were very happy and surprised to see Lipofectamine® 3000 reagent provide a more than 10-fold difference in transfection efficiency in our difficult-to-transfect cell line. (thermofisher.com)
  • For all practical purposes, a viral vector is a genetic element wrapped in its own transfection reagent, which makes transfection titrations possible (e.g., dosage responses) and highly effective. (biopharminternational.com)
  • In an R&D setting, this becomes preferable if the same reagent is transfected (for example, CAS9 delivery vectors) since all the work is in the generation of the viral stock, not in the infection," Carstens says. (biopharminternational.com)
  • The PureYield Plasmid Maxiprep System isolates transfection-quality plasmid DNA, providing a rapid method to purify up to 1mg of plasmid DNA from a 250ml bacterial culture. (promega.com)
  • The PureYield Plasmid Midiprep System isolates transfection-quality plasmid DNA, providing a rapid method to purify 100-200ug of plasmid DNA from a 50ml bacteria culture. (promega.com)
  • With your request submit a vector plasmid map or provide information about the backbone. (cincinnatichildrens.org)
  • Plasmid DNA (pDNA) is a critical delivery template for viral vectors. (repligen.com)
  • ORFs cloned in this vector will be expressed in mammalian cells as a tagged protein with an N-terminal DDK-tag. (origene.com)
  • Transfection of mammalian cells with an expression vector containing either TIF3 or TEF-1d cDNA resulted in overexpression of the encoded protein. (cdc.gov)
  • The development of commercially available transfection reagents for gene transfer applications has revolutionized the field of molecular biology and scientific research. (mdpi.com)
  • Transfection reagents developed and produced by Polyplus are key components in the manufacture of viral vectors. (sartorius.com)
  • Transfection efficiency was investigated via fluorescent microscope and flow cytometry. (cns.org)
  • All the pre- and post-modified PEI nanoparticles augmented the transfection efficiency of U87 cells, but the latter was better. (cns.org)
  • One of the post-modified nanoparticles (mole ratio: PEI/VTW=1/20) brought about the highest transfection efficiency among our polymers, even close to lipofectamine2000. (cns.org)
  • Vectors based on VTW peptide and PEI can improve gene transfection efficiency of U87 glioblastoma cells notably and specially. (cns.org)
  • However, cotransfection of multiple plasmids can cause low targeting efficiency in cells that are inefficient in DNA transfection. (nature.com)
  • In the present study the cellular tropism and transduction efficiency of seven AAV vector serotypes (AAV1, 2, 3, 4, 5, 6, 8) and an integration-deficient lentiviral vector were assessed for their ability to transduce corticospinal neurons (CSNs) following intracortical injection. (nih.gov)
  • The AFM results at the single polyplex level reveal likely connections between nanostructure and transfection efficiency of bioreducible polyplexes. (aiche.org)
  • You can achieve over 70% transfection efficiency in a cost-effective way. (thermofisher.com)
  • It delivers exceptional transfection efficiency with improved cell viability for the widest range of hard-to-transfect and common cell types including HEK 293, HeLa, LNCaP, HepG2, and A549 cell lines (Figure 1) . (thermofisher.com)
  • Far better transfection efficiency than Lipofectamine® 2000 when we compared them head-to-head. (thermofisher.com)
  • Spectacular increase in transfection efficiency and dramatic decrease in cell toxicity. (thermofisher.com)
  • Compared to adenovirus and AAV, retrovirus-based vectors show relatively low transduction efficiency in-vivo , but their advantage lies in their ability to integrate stably and confer long-term transgene expression in dividing tissues. (biopharminternational.com)
  • The biggest challenges in the clinical trials so far have been the limited gene transfer efficiency of the currently used gene vectors, the short duration of the transgene expression and the side-effects in viral-mediated gene transfer. (helsinki.fi)
  • The main purpose of this study was to investigate the effect of serum to the gene transfer efficiency of a nonviral vector polyethyleneimine PEI22K and the combination of PEI22K and cationic liposome Dosper in vitro in the SMC-cells. (helsinki.fi)
  • The potential synergistic increase in the transfection efficiency of PEI22K/Dosper combination was also studied. (helsinki.fi)
  • The secondary goal in this study was to develop an in vitro model which could be used to predict the gene transfer efficiency of gene vectors in vivo. (helsinki.fi)
  • The combination of PEI22K and Dosper resulted in a synergistic increase in the transfection efficiency in serum-free transfection. (helsinki.fi)
  • 1-10% serum concentrations did not significantly affect PEI22K`s transfection efficiency, but dramatically decreased the efficiency of PEI22K/Dosper combination. (helsinki.fi)
  • Products offered include viral vectors produced by transient transfection method or in stable producer cells, vector purification, and non-GMP quality testing. (cincinnatichildrens.org)
  • The Center is staffed by a team of experts in vaccine and viral vector production, purification and analytics. (repligen.com)
  • In addition, the plasmids for the second-step reaction retained the structure of a CRISPR/Cas9 expression vector harboring single gRNA and Cas9 expression cassettes and can therefore be used separately for single gene targeting. (nature.com)
  • In this study, we constructed plasmids to create all-in-one vectors expressing two to seven gRNAs with Cas9 nuclease or nickase ( Figure 1 ). (nature.com)
  • The VVC provides the standard packaging plasmids, the investigator provides the viral vector to be used for the production run. (cincinnatichildrens.org)
  • Transient and stable vector transfection: Pitfalls, off-target effects, artifacts. (nih.gov)
  • Using conformation-specific antibody staining and flow cytometry we report the surprising result that despite obtaining high transfection efficiencies and nominal vector yields in our model system, only 5-10% of cells appear to produce measurable levels of assembled AAV capsids. (cytovax.com)
  • Furthermore, we suggest that the flow cytometry assay used here to quantify productive cells may be a useful metric for future optimization of transfection-based AAV vector manufacturing platforms. (cytovax.com)
  • Recombinant adeno-associated viral (AAV) vectors and integration-deficient lentiviral vectors are promising therapeutic delivery systems for gene therapy to the central nervous system (CNS). (nih.gov)
  • Furthermore, it was shown that neither AAV nor lentiviral vectors generate a significant microglial response. (nih.gov)
  • Here we describe how microRNA-regulated lentiviral vectors can be used to visualize specific cell populations by exploiting endogenous microRNA expression patterns. (lu.se)
  • We provide detailed protocols for the generation of lentiviral vectors, the transduction of hES-cells, and their differentia- tion into neurons. (lu.se)
  • She has over 20 years' experience in scalable biologics and gene therapy manufacturing of therapeutic products, viral vectors and proteins for gene therapy. (repligen.com)
  • They are easy to culture and are commonly used in transfection and large-scale production of therapeutic proteins. (thermofisher.com)
  • Some vectors, such as adenoviruses, have a fairly broad tropism, and the target range is often modified by engineering part of the envelope proteins. (biopharminternational.com)
  • The expression cassettes of the gRNAs are tandemly ligated into a single vector using the Golden Gate cloning method. (nature.com)
  • The AAVS1-Targeting Knock-in HR Donor Vector 2.0 AAVS1-SA-puro-EF1α-MCS contains a multiple cloning site (MCS) downstream of the EF1α promoter, enabling constitutive expression of your gene-of-interest from the AAVS1 locus. (reportergene.com)
  • The expression of the target gene is regulated only in the cells with double transfection by the two vectors. (news-medical.net)
  • Here, we prepared a novel nanoparticle made by polymers which may solve the problem of targeting transfection into glioblastoma cells. (cns.org)
  • We further demonstrated successful examples of multiple targeting including chromosomal deletions in human cells using the all-in-one CRISPR/Cas9 vectors constructed with our novel system. (nature.com)
  • This finding implies that considerable increases in vector titer could be realized through increasing the proportion of productive cells. (cytovax.com)
  • En face co-immunostaining of the mouse aortic arch revealed a low level of PDCD4 in endothelial cells undergoing pulsatile shear stress. (plos.org)
  • In both cases, cells may be treated either with a viral or non-viral vector carrying the corrected copy of the gene. (biopharminternational.com)
  • Carstens points out that the greatest limitation of viral vector systems is their tropism, that is, the type of cells that a virus will naturally infect. (biopharminternational.com)
  • 3) VTW-modified nanoparticles is one kind of promising vector which has a potential to improve therapeutic effect of modern strategies. (cns.org)
  • Thus, we review application of AAV and EV-AAV vectors for DMD as novel therapeutic strategy. (cytovax.com)
  • Our viral vector experts work with our clients to identify and evaluate improvements to current processes in order to streamline therapeutic production, address risk, and accelerate cell and gene therapies to the clinic. (sigmaaldrich.com)
  • However, it is difficult to capture the complexity of transcript isoform variance using these libraries, and large cDNA sequences are often difficult to clone into size-limited viral expression vectors. (cdc.gov)
  • To make the most of the multiplexity of CRISPR/Cas9, we established a system for constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 nuclease/nickase expression cassette. (nature.com)
  • Here, we report an easy and efficient construction system for all-in-one CRISPR/Cas9 vectors expressing Cas9 protein and up to seven gRNAs. (nature.com)
  • In Cas9 nuclease-mediated genome editing, we constructed an all-in-one vector expressing Cas9 nuclease and seven gRNAs and targeted seven genomic loci. (nature.com)
  • To establish an all-in-one vector system, we modified the pX330 vector, originally developed by the Feng Zhang laboratory 4 , 8 , containing a single gRNA expression cassette and a Cas9 nuclease expression cassette. (nature.com)
  • Even though gene knock-outs can result from DSBs caused by Cas9 alone, SBI recommends the use of HR targeting vectors (also called HR donor vectors) for more efficient and precise mutation. (reportergene.com)
  • The core allows investigators to focus on their experiment rather than to spend time on trying to optimize the production of viral vectors themselves. (cincinnatichildrens.org)
  • Discover how we leverage our over 25 years of experience, knowledge and expertise in viral vector development and manufacturing to provide a de-risked, reproducible and streamlined path to clinical and commercial manufacturing through our viral vector platforms. (sigmaaldrich.com)
  • Join this webinar to learn how our Accelerator℠ process development services team helps our customers develop reproducible and scalable processes for viral vector manufacturing to meet clinical and commercial market demand in very short timelines. (pall.com)
  • Adding serum into transfection medium produced fewer influence to modified PEI nanoparticles compared with lipofectamine2000. (cns.org)
  • This is partly due to the interaction between serum components and nonviral vectors. (helsinki.fi)
  • Oligonucleotides corresponding to each target sequence are annealed and inserted into BpiI-digested pX330A or pX330S vectors (STEP 1). (nature.com)
  • Nonviral gene transfer agents have so far been less efficient in vivo than the viral vectors. (helsinki.fi)
  • How to correlate physiochemical attributes with transfection and toxicity data remains a major scientific challenge. (aiche.org)
  • The application of recombinant DNA technology to fields such as drug discovery and development has led to an increased need for high-throughput transfection. (qiagen.com)
  • By vaccinating hamsters with a recombinant parainfluenza virus vector, Buchholz et al. (cdc.gov)
  • The constructed vectors harboring single gRNA expression cassettes are then assembled into an all-in-one vector harboring multiple gRNA cassettes using the Golden Gate assembly method (STEP 2). (nature.com)
  • The TAV vector harboring 100 nucleotides of the phytoene desaturase ( PDS ) gene caused photobleaching phenotypes and a reduction in CsPDS expression in C. seticuspe . (cytovax.com)
  • Forward primer to sequence targets cloned in expression vectors. (origene.com)
  • When you want to take advantage of the robust and reliable expression that's possible from the AAVS1 Safe Harbor Site, SBI offers a range of second generation (2.0) AAVS1 Safe Harbor Site Targeting HR Donor Vectors that are designed minimize off-target integration. (reportergene.com)
  • We generated IRS1 expression vectors harboring p.S668T, p.D674H and double mutants. (scielo.br)
  • The collaborative research team led by Professor Tadashi ISA, Project Assistant Professor Masaharu KINOSHITA from The National Institute for Physiological Sciences, The National Institutes of Natural Sciences and Fukushima Medical University and Kyoto University, developed 'the double viral vector transfection technique' which can deliver genes to a specific neural circuit by combining two new kinds of gene transfer vectors. (news-medical.net)
  • Viral vector systems are gutted viruses where the packaging signals (e.g., the genetic signals that target the wrapping into the viral particles) are fused to the genes of interest (the payload), and the remaining necessary elements are placed on separate genetic elements, often three or more, says Carsten Carstens, senior scientist, R&D, Agilent Technologies. (biopharminternational.com)
  • Repligen end-to-end solutions help overcome key challenges in pDNA and viral vector manufacturing, with hands-on process and implementation consultation from global Field Applications Specialists and recognized gene therapy, oncolytic and vaccines industry experts. (repligen.com)
  • The Viral Vector Core offers production of research-grade vectors, generation of stable producer lines, and non-GMP quality control testing including vector titer by functional assay FACS or PCR, endotoxin, mycoplasma, and USP sterility testing. (cincinnatichildrens.org)
  • All four steps in the transfection process were optimized and our most advanced lipid nanoparticle technologies were combined. (thermofisher.com)
  • The ORF cloned in this vector can be expressed as a tagged protein with a c-terminal GFP tag. (origene.com)
  • The scientists constructed a CRISPR-Cas 9 vector that targeted the DJ-1. (isaaa.org)
  • Depending on the application, the ideal vector either has an extremely broad tropism or extremely narrow if the objective is gene therapy of a very specific cell type (in this case this is a safety precaution)," Carstens elucidates. (biopharminternational.com)
  • The process of changing the tropism of the vectors is referred to as pseudotyping. (biopharminternational.com)
  • Our wide range of technologies and extensive knowledge of bioprocessing platforms make us uniquely qualified to offer end-to-end support for viral vector processes. (pall.com)
  • Millipore ® CTDMO Services delivers expertise and flexible solutions for viral vector development and manufacturing to advance cell and gene therapies from preclinical through commercial production. (sigmaaldrich.com)
  • Our viral vector experts are dedicated to helping your advance your viral gene therapy from pre-clinical through commercial production, backed by a superior regulatory track record. (sigmaaldrich.com)
  • The core works closely with the GMP-compliant Vector Production Facility and is able to transfer technology for further development and scale-up and ensure a seamless transition from preclinical studies to early-phase human gene therapy clinical trials. (cincinnatichildrens.org)
  • The clever design of our second generation AAVS1-Targeting HR Donor Vectors limits off-target integration for highly-specific targeting of the AAVS1 site. (reportergene.com)
  • each of the vectors has its own advantages, limitations, and range of applications," Vicaire says. (biopharminternational.com)
  • Agilent vectors, for example, are suitable and compatible with generation-four packaging systems (i.e., the latest version of packaging systems). (biopharminternational.com)
  • Synthetic delivery vectors based on self-assembly of DNA and polycations (polyplexes) continue to gain strength as viable alternatives to viral vectors. (aiche.org)
  • Viral vectors show promise as a delivery mechanism for gene therapy, but which virus types are commercially viable? (biopharminternational.com)
  • The TTAA-specific transposon PiggyBac™ is a mobile genetic element that efficiently transposes between vectors and chromosomes via a 'cut and paste' mechanism. (bioscience.co.uk)
  • They can draw on experience from multiple gene therapy viral vector projects to collaborate with you. (pall.com)
  • All OriGene's TrueORF clone can be shuttled into this vector via PrecisionShuttling, a simple cut-and-ligation process. (origene.com)