We narrowed to 302 results for: expression vector

...derivatives of HEK-293, the most commonly used line expresses SV40 large T antigen and is thus dubbed HEK-293T...packaging cells’ for viral production because they express the SV40 large T-antigen. This allows them to replicate...receptors and ion channels are often exogenously expressed and studied in HEKs due to the cell line’s ability...produce these proteins and its relatively low expression of native channels.  No matter what your research...factors that could skew research results (gene expression, growth rates, etc.). Thus, it’s important to...Platform to Produce Recombinant Proteins and Viral Vectors. Front Bioeng Biotechnol. (2021) (9):796991. DOI...10 Tips for Mammalian Cell Culture Adenoviral Vector Production and Troubleshooting Additional resources...

...because they can be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs... plant, and insect cells, they are most often expressed from a plasmid in bacteria (Ahmed et al., 2012...2012). One challenge of bacterial expression is the proper formation of the disulfide bond between the VL...requires an oxidizing environment to form. When expressed in the non-oxidizing cytoplasm of bacteria, scFvs...more of these approaches are often used for scFv expression in bacteria:  targeting to the oxidizing periplasm...to allow for proper disulfide bond formation.  expression in strains of bacteria that are redox mutants...mutants so they have a more oxidizing cytoplasm. expression in the presence of molecular chaperones that help...

...recognized by the Sep-tRNA. 3. An expression plasmid encoding the ORF to be expressed Finally, the researcher needs...ORF of interest is cloned into one of these expression vectors, the codon for the amino acid to be replaced...non-phosphorylated serine at the TAG codon of the same expression vector used for phosphoserine incorporation (6). ...bacterial host strain for protein production, and an expression plasmid encoding the open reading frame (ORF)...encoding their open reading frame of interest. Two expression systems have been verified for use by the Rinehart...TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can ...using standard protein purification strategies; expression should be optimized for parameters such as induction...

...codon optimized specifically for expression in yeast. These tagging vectors are based on previously described...interest. These fluorescently tagged genes are expressed under native conditions and allow scientists to...described pFA6a-link vectors and include a Kan, SpHIS5, or CaURA3 selection marker. Lee et al.(1) assessed...from these yeast-optimized fluorophore tagging vectors for your single or multi-color imaging experiments...artifacts sometimes associated with plasmid-based overexpression. Tim Formosa, at the University of Utah, has..., including full and partial gene deletion, overexpression (by promoter substitution), and tagging at ...

...guide to viral vectors Find all of our viral vector blog posts Download our viral vectors 101 eBook Additional...pressure on the production of retroviral vectors: Enhancement in vector stability." Biotechnology and bioengineering...been actively investigating ways to improve viral vector production. Whether redesigning viral packaging...membrane When packaging adeno-associated viral vectors (AAV) not all serotypes are equal; while some serotypes... of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...we believe that this approach can be adapted by vector cores and laboratories alike as an easy and inexpensive...Efficient serotype-dependent release of functional vector into the culture medium during adeno-associated...

... while we think of the AAV vector genomes as containing just the expression cassette that we want, it ... our viral vectors related blog posts Read our blog post on AAV titers Browse viral vectors protocols ... platform to accommodate adeno-associated viral vector (AAV) samples and created a simplified process,...methods for AAV quality control Historically, AAV vector QC has been sparse including only titration of ... of quality control is sufficient but for viral vector core facilities producing a large number of viral...demonstrated that NGS is an effective tool to confirm AAV vector genome identity. These studies used SMRT sequencing... simplified approach, we sequenced over 100 AAV vector preparations and showed that VGS can quickly and...

...easy process. The cloning capacity of lentiviral vectors (~8.5 kb of insert) and the reduced titer associated...providing lentiviral preps of select lentiviral vectors from the repository for you. We perform test infections...infections with all of our viral preps and host our expression data (western blots or fluorescent microscopy...nickase for genome editing. You can also enhance expression of your gene of interest with a variety of Cas9...therefore, must be used to infect a cell line already expressing Cas9. As John Doench (Broad Institute and member...Page Check Out Our CRISPR Guide Pages Find Viral Vectors ...

...pools are available in two vector formats: all-in-one (lentiCRISPRv2, expressing both Cas9 and an individual...individual gRNA from this single vector) and guide-only (lentiGuide expressing only an gRNA, for use with cells...plasmids will also be useful for imaging or gene expression assays necessitating the screening of individual...

...modular vector assembly process designed for a quick, plug-and-play approach to cloning CRISPR vectors (McGee...for each module, generate the vector file, and quickly clone their vector of interest. And it truly is ...destination vectors for various delivery methods, along with compatible inserts. All destination vectors contain...heavy lifting out of CRISPR vector design, making it easier to get the vectors that are just right for your...Fragmid destination vectors currently available. Table 1: Fragmid destination vectors Organism Delivery...designing a CRISPR vector the Fragmid way!    Figure 1: Selecting your vector type on the Fragmid...plasmid and generating the vector file on the Fragmid website.   If the vector is to your liking, your...

...2016 were CRISPR plasmids), but so are lentiviral vectors, many of which are used to deliver Cas9 and other....G from Didier Trono’s lab! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000... 6,000 times (over 800 orders in 2017) and expresses the VSV-G envelope protein under the control of the... our Viral Production Process Find other Viral Vectors ...

... injection of an AAV expressing a ChRger. In both experiments, neurons expressing ChRgers produced stronger... what was seen with ChRger2 expressing mice, while ChR2(H134R) expressing mice did not have this self-...Addgene.org Browse all viral vectors available at Addgene Visit our viral vectors guide Find our chemogenetics...to fire when light is shined on them, but the expression of light-gated ion channels such as channelrhodopsins...When light shines on excitatory channelrhodopsin-expressing neurons, the channelrhodopsins respond by opening...sequence and structures that determine membrane expression and plasma membrane localization of transmembrane...and ChRger3, were then characterized further by expression in cultured neurons and mouse brain slices collected...

...Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...that overexpress genes of interest; more recently we have also been using Addgene CRISPR vectors to generate... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...and PCR, depending on the level of endogenous expression. We highly recommend the use of an electronic...ampules each of 4 clones with different levels of expression of our gene of interest. Advantages of the agarose...isolate individual cell clones with different expression characteristics from a population of transfected...Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org Check Out Our Protocols...

...multiplexed gRNA expression (Wong et al., 2016), in addition to the combinatorial expression of other genetic...CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12 ...below and pool-clone them into the pAWp28 storage vector (Figure 1): Forward oligo: 5’- CACCGNNNNNNNNNNNNNNNNNNNGTTTGGGTCTTCGAGAAGACCTATTCXXXXXXXXC...barcode, respectively. Once cloned into the storage vector, the pooled, barcoded gRNA library is now ready...pAWp12 lentiviral vector should retain the diversity in your gRNA library, but this should be verified by NGS...

...with GeneWeld vectors GeneWeld from Jeffrey Essner’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins... tools for your research! This time: GeneWeld vectors to create knock-ins CasX Drug inducible CRISPR/...cargo (ex: fluorescent protein gene) on the donor vector. Therefore, targeting requires two sgRNAS: one ...site and a universal sgRNA that targets the donor vector sequence. Homology arms that flank the donor plasmid... Read the paper in bioRxiv Find the GeneWeld vector kit CasX, a small Cas protein, is active in E....active and inactive CasX for E. coli and human expression. Read the paper in Nature Find the CasX plasmids...

...steps in the coming weeks. Expression of the optogenetic constructs Expression of your desired optogenetic...promoter-directed expression alone is extremely limited. Therefore, highly expressing, generic neuronal...with an appropriate mouse line expressing a particular cre, viral expression of your opsin is no longer required...Finally, expression of the opsin in a mouse line will likely be less than that of viral expression but may...stimulation. Testing opsin expression  Regardless of the method of expression of your opsin gene, it is...extremely high expressing promoters). Conduct a time course experiment in which you monitor expression at various...essential to test the expression of your opsin. Thankfully the Deisseroth constructs coexpress a fluorescent ...

...manner. They designed the pXPR_053 lentiviral expression vector, which contains an sgRNA insert site and a...pXPR_053. They expressed this construct in bone marrow stem cells from Cas9-expressing mice, and used ... microbe travels through a host organism, the expression of some genes can change depending on the location...reporter gene (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids .... 2018. PubMed PMID: 29300010. miRNA reporter vectors shed new light on mechanisms of miRNA-mediated ...important regulators of post-transcriptional gene expression across the majority of cellular pathways. Despite...target site. One fluorescent protein (EBFP2) is expressed in an unregulated fashion and thus a marker for...

...researchers with many different ways to control gene expression. Get the full scoop from Addgenie A. Max Juchheim... rundown of some of the strains you can use to express your protein of interest. 4. Plasmids 101: Blue-White...Blue-White Screening Getting a lot of empty vector contaminants in your cloning experiments and tired of...CRISPR/Cas9 and AAV Adeno-associated viral (AAV) vectors are useful for in vivo gene delivery, but have ...Browse Our CRISPR Resources Learn More about Viral Vectors Browse the Repository  ...

... a GAL-4 based bipartite expression system that can regulate gene expression in C.elegans. In this system...CRISPR techniques and new tools for conditional expression and degradation. These workshops not only highlighted...elegans open resources The C. elegans Neuronal Gene Expression Map & Network (CeNGEN) The CeNGEN goal is to ...to establish a comprehensive gene expression atlas of every C.elegans  neuron. Scientists are currently...system a promoter controls the expression of a GAL-4 driver that can bind to activation sequences upstream...upstream of genes to stimulate expression (Wang et al., 2016). In 2018, the lab expanded on this tool by ...cells that have both halves of cGAL can drive expression of your gene of interest.  SapTrap Tool Kit for...

...four reprogramming factors can be delivered and expressed in multiple somatic cells via various methods....transdifferentiated in vivo to pancreatic β-like-cells with the expression of pancreatic and duodenal homoeobox gene1 (PDX1...transdifferentiation is usually carried out by expressing transcription factors specific to the lineage...and in vitro methods are similar except that the vectors carrying the transdifferentiation factors are directly... (iPSCs) by temperature-sensitive Sendai virus vectors. Proc Natl Acad Sci U S A, 2011. 108(34): p. 14234...., Human induced pluripotent stem cells free of vector and transgene sequences. Science, 2009. 324(5928...mouse induced pluripotent stem cells without viral vectors. Science, 2008. 322(5903): p. 949-53. PubMed PMID...

...determined protein expression levels from different promoters, and inhibited gene expression using CRISPRi....transduced at levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo ...parts, selectable markers, transformation acceptor vectors, and sequences for genomic integration by homology...use. This kit is designed for use with acceptor vectors from the MoClo Toolkit from Sylvestre Marillonnet...increases temporal and spatial precision of gene expression and reduces toxicity as compared to previous ...zebrafish model, ubi:zebrabow. In this system, cells express various fluorescent proteins controlled by Cre ...depending on the combination of fluorescent proteins expressed. CreLite-mediated recombination was found in both...