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We narrowed to 263 results for: Cre expression vector

Showing: 161 - 180 of 263 results

  1. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ...can also detect expression of toxin mRNA from Clostridium difficile. As mRNA expression cannot be identified...2009). The vector retains the oriT for delivery into recipients by conjugation. The labs screened for the...upon the Yeast Toolkit by replacing the backbone vector’s high-copy number bacterial ColE1 origin with the...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...of cells containing the particle exponentially decreases over time and the number of elapsed generations...

  2. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ... 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 ...consider scaling up by either increasing the number of 10 cm2 dishes or increasing the vessel size. Opting ...

  3. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...lentiviral transfer vectors to create the CRISPR library. CRISPR libraries have been created to knock out, ...deliver a pooled population of gRNAs to Cas9 expressing cells. Pooled lentiviral CRISPR libraries (referred...population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within the genome...) to be used to generate lentivirus (B). Cas9-expressing cells or wild-type cells are treated with lentivirus...genome-wide screens? The goal of a genome-wide screening experiment is to generate and screen a population...the steps involved in a CRISPR screen? Performing a forward genetic screen using CRISPR libraries is a multi-step...sorting and screened for a particular phenotype. For example, mutant cells can be used in drug screens to identify...

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

    Type
    Blog Post
    Published
    June 2, 2020, 1:15 p.m.
    ...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:/... Figure 1: Initial steps in the CRISPR-pooled screening using sgRNAiBAR. Image from Zhu et al., 2019.  ... Peking University has developed a new CRISPR screening method that makes use of guide RNAs with internal...

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

    Type
    Blog Post
    Published
    Dec. 3, 2019, 11:38 a.m.
    ...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...more about this!). I’ve assembled a series of screenshots to highlight changes to both the design and functionality...NeuroMab Collection for details). The following screenshots show the range of different terms we can use ...

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

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...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,...

  7. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ...Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery...SpCas9) and a gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of...activation. Split inteins have also been used to express base editors. References Chew WL, Tabebordbar...HBB and CCR5 genes in human cells using compact expression cassettes. Sci Rep 5: . https://doi.org/10.1038...genome engineering: An adeno-associated viral (AAV) vector toolbox. Biotechnology Journal 9:1402–1412 . https...Editing: Strategies and In Vivo Delivery by AAV Vectors. Cell 181:136–150 . https://doi.org/10.1016/j.cell...

  8. Same Addgene, New Look - Why We Redesigned Our Homepage & Mascot

    Type
    Blog Post
    Published
    Dec. 5, 2019, 1:49 p.m.
    ...’ the homepage is still important. It needs to express value, clearly communicate exactly what a site ...wanted users to be able to find plasmids and viral vector tools more easily. In the redesign, the new search...One big feature upgrade? Blugene now has a full expression. You’ll mostly see a friendly smile, but occasionally...you come to Addgene to find the plasmid or viral vector for your next experiment, that you can find what...Addgene.  Addgene's new homepage While users are increasingly accessing websites other than through the ‘front...

  9. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    Published
    Sept. 6, 2017, 1:01 p.m.
    ...homology. Producing yeast expression cassettes Traditionally, generating expression cassettes for yeast transformation... mutations Cloning sgRNAs Production of Yeast Expression Cassettes Generating constructs with multiple...PCR to generate the mutation and linearizing your vector (preferably by PCR) for DNA assembly. Further, ...simple as mixing fragments together with linearized vector for seamless joining of multiple mutations spread...physically joining multiple fragments of DNA to create a synthetically designed DNA sequence. There are...fragment joining, but this is fine for its goal of creating a choice of standardized constructs and tools ... regions. Golden Gate Assembly also allows the creation of standardized DNA constructs, but its use of...

  10. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...same strain. MazF expression is under control of the AOX1 promoter whose expression is induced by methanol...that are used to drive high protein expression in vivo. Expression plasmids are often lost without plasmid...Kaper used the hok/sok system to create strains that can stably express antigens in sufficient quantities...as a counterselectable marker in the destination vector. Successful cloning occurs when the ccdB gene is...methanol. Once linearized, the delivery vector can be integrated into the genome to introduce the genetic...plasmid R1: Sok antisense RNA regulates hok gene expression indirectly through the overlapping mok gene."...James B. Kaper. "Use of stabilized luciferase-expressing plasmids to examine in vivo-induced promoters...

  11. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...specific cell types is where viral vectors become invaluable. Viral vectors are mechanisms by which genetic...Resources on the Addgene blog Viral Vectors 101: Chemogenetics Viral Vectors 101: Optogenetics Fluorescent...Sensors This system consists of a genetically expressed enzyme, typically on the cell surface, which can...fluorescence or voltage indicators.   Viral Vectors and Voltage Indicators As previously mentioned,...hydrolyzes a precursor dye to make it more lipophilic, increasing its affinity for the membrane, where it will ...

  12. What's New in CRISPR - March 2020

    Type
    Blog Post
    Published
    March 10, 2020, 1:15 p.m.
    ...fluorescently labeled piggyBac vectors to deliver uniform and sustained expression of multiplexed gRNAs in human...CRISPRa depended on continued expression of dCas9 and gRNAs whose expression varies depending on transgene...The lab expressed both dCas9 and the ctrA sgRNA and saw that the levels of ctrA mRNA decreased after 20...demethylation Optogenetic repressors of gene expression in yeast Megan McClean’s lab generated an optogenetic...optogenetic toolkit that controls repression of gene expression in Saccharomyces cerevisiae. The system consists...the nucleus where dCas9-Mxi1 can repress gene expression. In the dark, the NLS is inaccessible and the... in Caulobacter cells for ctrA gene expression using various dCas9. Image adapted from Guzzo et al., 2020...

  13. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ...structures in live cells. Four individual expression constructs were created, each containing a unique fluorescent... construct. This strategy can be used to build vectors with diverse functions, and thus, is used in many...localization, as well as cellular processes like gene expression and cell migration (Sladitschek and Neveu, 2015...enhancers, polyadenylation signals, inducible gene-expression sequences, and more. By combining these components...cell cycle dynamics, titrate inducible transgene expression, or a variety of other clever applications. ...flanked with a CMV promoter (to drive high-level expression) and a polyA signal (to terminate transcription... HeLa cells show robust labeling, with strong expression and detection of each fluorophore at the respective...

  14. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ...This will be highly dependent on vector dose. The color diversity in Vector Assisted Spectral Tracing (VAST...drop in transduction coverage compared to just one vector.   Q2: Do you think it is possible to make an efficient...this scenario, you may be best served by a dual vector, split intein approach.   Q3: What’s the best method...for model organisms.    Q11: How consistent is expression is when injecting CAP-B10 or CAP-B22 in marmosets...Plasmids Adenovirus Guide Page Subscribe to Viral Vectors Blog Posts ...flexibility of AAV genome design in remarkably creative ways. The continued development of more and smaller...

  15. Five Popular Model Organisms

    Type
    Blog Post
    Published
    April 11, 2019, 12:39 p.m.
    ...lab who created a set of modular vectors that can be used to optimally control gene expression in specific...spatiotemporal expression of a gene of interest. This type of control over gene expression is possible in... al., 2009). By injecting mice with retroviral vectors that harbored different mutations found in leukemia...that uses heat induction to control the temporal expression of different transgenes, the Stainer group determined...micro-injecting DNA into the worm, such as animals that express fluorescent markers (Mello et al., 1991). Genes...interference (RNAi) by feeding the worm bacteria expressing specific plasmids (L4440) that contain your gene...lifetime. Thus any genetic modification, such as expression of fluorescent protein or disruption of a gene...

  16. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    Published
    Dec. 6, 2018, 1:34 p.m.
    ...switch to be expressed. In this case, the gene encodes the LacZ𝛼 protein. Once expressed, LacZ𝛼 complexes... RNAi. Recently the Vellai lab has created an improved RNAi vector that leads to significantly stronger... RNAi is commonly achieved by feeding E. coli expressing dsRNA to C. elegans. These dsRNAs target specific...for mutant genetic backgrounds. The standard RNAi vector, L4440, places a target gene between two inverted...mutant animals that are genetically altered to overexpress endogenous RNAi pathways. These mutations however...non-specific nucleotides into the dsRNA leading to increased RNAi efficiency. Find the T444T plasmid here... used yeast display-based directed evolution to create APEX2. They have now applied the same directed ...

  17. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...codon site of interest can be cloned into an expression vector with Gibson assembly. While this is a comprehensive...relies on the generation of a plasmid library that expresses all desired variants of a protein. Applying a ...selective pressure winnows the pool down to plasmids expressing variants with optimal function. High-throughput...is generated by transforming a backbone phagemid vector into an E. coli strain that is deficient in dUTPase...2 instances where it does. 1) When the backbone vector has multiple BbvCI sites. If these sites have opposite...polymerase, 2) increased PCR extension time, 3) increased concentration of MgCl2, 4) increased concentration...PCR reaction is created by making some or all of the following modifications: 1) increased concentration...

  18. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...those optogenetic controls, gene expression levels reliably increased in vitro by a factor of 10 to 20...functions, Konermann needed tools to manipulate the expression of specific genes in the brain in reversible ...Zhang showed they could package LITEs in viral vectors and genetically target them to specific cell populations...demonstrated use of the LITEs to control gene expression in mouse neurons and in the brains of living ...system could be by applying it to altering gene expression via epigenetics with the addition of histone ...different effectors. I’d encourage people to get creative with them for their own uses.” Konermann said ...found here.) So tell us, readers, how will you get creative with LITE? Reference: Silvana Konermann et al...

  19. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    Published
    March 5, 2019, 1:08 p.m.
    ...promoter is expressed. Cre-mediated deletion of one or several of these genes permits the expression of genes...copy of the transgene expressing either YFP or CFP, and cells that do not express Cre fluorescing with the...activate the expression of a different fluorescent protein. Therefore, each cell could express a unique combination...fluorescent protein genes, Cre recombination is required before fluorescence can occur. Cre recombinase can be...developing fish via Cre protein microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier...any cell type when crossed with mice expressing the appropriate Cre allele. The Confetti mouse contains...so that XFP are only expressed following a recombination event. Tissue-specific Cre recombination in this...

  20. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ... are transiently expressed using either AAV backbones or species-specific expression plasmids. You can...cloning strategies for constructing shRNA expression vectors. BMC Biotechnology, 6(1). https://doi.org...processing and silencing pathways. Created on BioRender.com.   Expressing shRNAs using plasmids The design...sequence. Created on BioRender.com. When it comes to the backbone, shRNAs are most often expressed constitutively...option for altering the expression of specific mRNAs. shRNAs are a quick way to create stable knockdown cells...be applied to. When it comes to altering gene expression, CRISPR technologies have gradually taken over...processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the endogenous miRNA...
Showing: 161 - 180 of 263 results