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We narrowed to 407 results for: expression vector

Showing: 161 - 180 of 407 results

  1. A Primer on Optogenetics: Introduction and Opsin Delivery

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    Blog Post
    ...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 ...

  2. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    ...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...

  3. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    ...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  ...

  4. Open Resources and Plasmid Tools For Studying C. elegans

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    Blog Post
    ... 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...

  5. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    ...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...

  6. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...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...

  7. Working with Nuclear Receptors

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    Blog Post
    ... a vast number expression and reporter plasmids and the free exchange of these vectors between laboratories... ligands remain unknown. Together, NRs govern expression of genes involved in a broad range of reproductive...efficient and timely activation or repression of gene expression (see www.nursa.org for a curated list of coregulators...ligand, cell type, and promoter, organizing the expression of functional groups of genes involved in the...Our Q-PCR datasets, for example, document the expression patterns of nuclear receptors across multiple...researchers with a one-stop shop for plasmids and vectors, and making them available to all researchers for..., B. W. (2002). Combinatorial control of gene expression by nuclear receptors and coregulators. Cell 108...

  8. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...feature of this system is a lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient knockout...model in vivo. In this screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8...robustness of the system and utility of the pRDA_526 vector. Moreover, the screen identified putative novel...

  9. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    ... cDNAs into a single consensus lentiviral expression vector and attached a DNA barcode to each to make...cDNAs) representing core nodes that, when over-expressed, constitutively activated or inactivated the pathway...other researchers, and cloned them into lentiviral vectors to generate a kit which is now available at Addgene...

  10. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...bacteria that have taken up this vector, but it won’t tell you whether the vector contains the gRNA insert or...X-gal allows you to identify cells that took up the vector and distinguish between plasmids that contain the...𝛽-galactosidase. Complementing this mutation by expressing this deleted portion, for example on a plasmid...functional 𝛽-galactosidase. Blue-white screens rely on vectors where the gene of interest is inserted into the..., you will see two bands: one representing your vector backbone and the other representing the insert....doesn’t contain the insert, you will just see the vector backbone band. Or, you can choose an enzyme that...specific to your insert or by using primers that are vector specific but amplify a potential insert. If you...

  11. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...producing infectious virus (similar to lentiviral vectors) because they do not encode all of the necessary...plasmids allows researchers to partially control expression levels of each gene by altering the amount transfected...RNA polymerase gene or using cells that stably express T7 (e.g. BSR T7/5 cells) (1). We hope that minigenomes...on the Addgene Blog Read Other Posts on Viral Vectors Check out Our Scientist's Guide to Ebola Contribute... viral vector...

  12. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...can also detect expression of toxin mRNA from Clostridium difficile. As mRNA expression cannot be identified...upon the Yeast Toolkit by replacing the backbone vector’s high-copy number bacterial ColE1 origin with the...origin with broad host-range (Meyer, 2009). The vector retains the oriT for delivery into recipients by...resistance markers, promoters, and inducible gene expression systems in bee gut microbiome species such as... or a bacterial microcompartment protein. The expression of this fusion is under control of the arabinose-inducible...Cells are first induced with arabinose to begin expressing the SAP-RFP fusion. Then, the cells are washed...

  13. Scientists Map the SARS-CoV-2-Human Interaction Network

    Type
    Blog Post
    ...one are in a pLVX vector background, so you can package them into lentiviral vectors if you need to. Additionally...selection. Expression is driven by a fairly strong EF1alpha promoter. We have verified expression of correctly-sized...2 virus-human interaction network in which we expressed 26 of the 29 SARS-CoV-2 viral proteins in human...Western Blot for 25 constructs, and confirmed expression via mass spec for two of the remaining viral ...

  14. Overcoming the Challenges of Lentiviral Production

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    Blog Post
    ... While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges... a usable titer, or fine-tuning selection and expression in your target cell line, researchers often find...transcriptional activation system to drive high levels of expression of certain viral components; according the manufacturer...determination of the packaging limit of lentiviral vectors. Hum Gene Ther. 2001 Oct 10;12(15):1893-905. PubMed...Tips for titering your lentivirus Visit the viral vector featured topic page Resources on Addgene.org ...Guide Find lentivirus plasmids Browse all viral vectors ...

  15. AAV Titers: Where Do They Come From And What Do They Mean?

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    Blog Post
    ... viral vector preparations. Addgene has used this method to generally validate our viral vector preparation...of an Addgene AAV viral vector preparation after negative staining. Empty vector particles can be identified...darker than full vector particles. This image shows that the vast majority of the vectors consist of full...many ways to quantify the composition of a viral vector solution, and generally these methods measure different...infectious titer. The infectious titer of a viral vector stock can vary based on transduction target and...typically quantified by electron microscopy of a viral vector solution. Since this technique is intensive and...over 95% of viral capsids in representative viral vector preparations are genome-containing (Figure 1).  ...

  16. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    ...the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames and identified...plasmids expressed 26 of the 29 SARS-CoV-2 proteins which all but one are cloned into pLVX vectors (for lentiviral...provide synthesized SARS-CoV-2 expression plasmids available for many expression systems: bacterial, yeast,...C-terminal deletion in the spike protein to improve expression and pseudotyping efficiency. This plasmid was...enzymes. Plasmids in this toolkit include ones for expressing MMLV reverse transcriptase and Taq polymerase...-CoV infection and deposited plasmids for the expression of cathepsin S, L, and B. This became a great...human cells. Addgene’s mammalian target plasmids express these and other targets such as TMPRSS2, FURIN,...

  17. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...cells with sgRNAiBAR library containing lentiviral vectors the iBARs can be detected by next generation sequencing...the stop cassette is removed, the effector is expressed. By truncating one of the recombination sites ...calcineurin-dependent response elements (CDREs) and drives gene expression. Because calcineurin is required for virulence...4X CDRE repeat. In addition to showing that this vector is suitable for flow cytometry and fluorescence... single molecules in real-time. Find plasmids expressing tTALE-FP Shin et al., Sci. Rep. 2019. https:/...

  18. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...Addgene Resources on Addgene.org Find Yeast Expression Vectors Check Out Our Molecular Biology Reference...yeast species into the pYES2.1/V5-His yeast overexpression vector. This plasmid contains origins of replication... or S. cerevisiae. Students then determine if expression of the plasmid-encoded MET gene allows the transformed...cannot grow without uracil supplementation (6). Expression of MET/Met genes from the pYES2.1 plasmid is ...proteins, which can be used to confirm protein expression by western blotting in other experiments (Figure...Students then use replica plating to determine if expression of MET/Met genes from the plasmids allows the...Since the GAL1 promoter controls MET/Met gene expression, complementation should only occur when galactose...

  19. Hot Plasmids - October 2020

    Type
    Blog Post
    ...from other regions of the cell. By combining vectors expressing different fluorescent proteins targeting ...display (FingRs), Xue Han’s lab has developed AAV vectors to selectively visualize excitatory and inhibitory...excitatory and inhibitory synapses, respectively. Once expressed in the neuron, the FingRs bind to the endogenous.... The team engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular...proliferation in acute myeloid leukemia and saw that expression of H2B-FT consistently predicted the cell cycle...different cell types. In one example, H2B-FT was expressed in murine B cells (BaF3), which depend on IL-3...screening Applications (CHyMErA) Knockout Libraries express hybrid Cas9-Cas12a gRNAs under a single U6 promoter...

  20. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    ...categories Expression: Select the model system for your research (e.g. Mammalian) Vector Type: What type...Addgene is currently distributing 94 plasmids expressing validated antibodies for neuroscience research...example I wanted to find a lentiviral plasmid to express ubiquitin under control of a CMV promoter. Starting...but I can’t browse through 300+ plasmids. Under ‘Vector Types’ I selected ‘Lentiviral’ which eliminates...Identify a specific marker protein Use: Similar to “Vector Type” you can filter by experimental purpose (e.g...
Showing: 161 - 180 of 407 results