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We narrowed to 407 results for: expression vector

Showing: 181 - 200 of 407 results

  1. X-CHIME: Context Dependent Germline Knockout in Immune Cells

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    Blog Post
    ...transduction of Cas9-expressing (Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a ...utilizes custom mouse strains and lentiviral expression vectors (Table 1). The four systems Name Function...

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

    Type
    Blog Post
    ...’ 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...

  3. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ...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...packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...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...

  4. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...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...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...hok/sok system to create strains that can stably express antigens in sufficient quantities to generate an...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...

  5. Pushing the Limits of DNA Assembly

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    Blog Post
    ...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...

  6. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ...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,...

  7. AAV Q&A with Tim Miles

    Type
    Blog Post
    ...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 ...

  8. MXS Chaining

    Type
    Blog Post
    ... 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. ...subcellular structures in live cells. Four individual expression constructs were created, each containing a unique...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...

  9. Hot Plasmids: Winter 2025

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    Blog Post
    ...cell-derived neurons expressing ASAP5-Kv. C) Cultured rat hippocampal neuron expressing pan-membrane ASAP5...Each plasmid expresses all the components of the biosensor in optimal proportions from a vector backbone ...Figure 2), the authors developed a cost-effective expression and labeling approach using these constructs ...protein of interest can be inserted. B) Protein expression and labeling procedure. Adapted from Smith et..., and CLAP-tagged proteins for mammalian cell expression. These plasmids will be useful not only for future...recently developed a collection of BRET-based ONE vector G-protein Optical (ONE-GO) biosensors (Janicot ...

  10. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...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...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...of CRISPRi in Caulobacter cells for ctrA gene expression using various dCas9. Image adapted from Guzzo...lab developed a CRISPRi system to downregulate expression of specific Caulobacter genes using a catalytically...

  12. Five Popular Model Organisms

    Type
    Blog Post
    ...spatiotemporal expression of a gene of interest. This type of control over gene expression is possible in...created a set of modular vectors that can be used to optimally control gene expression in specific cells of... 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...

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

    Type
    Blog Post
    ...switch to be expressed. In this case, the gene encodes the LacZ𝛼 protein. Once expressed, LacZ𝛼 complexes... RNAi is commonly achieved by feeding E. coli expressing dsRNA to C. elegans. These dsRNAs target specific...Recently the Vellai lab has created an improved RNAi vector that leads to significantly stronger phenotypes...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...

  14. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...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...

  15. Let There Be LITE Plasmids

    Type
    Blog Post
    ...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 ...stimulation. Under those optogenetic controls, gene expression levels reliably increased in vitro by a factor...system could be by applying it to altering gene expression via epigenetics with the addition of histone ...

  16. Plasmids 101: Expressing shRNAs for RNA Interference

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    Blog Post
    ... 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...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...silencing pathways. Created on BioRender.com.   Expressing shRNAs using plasmids The design of shRNA plasmids... comes to the backbone, shRNAs are most often expressed constitutively, either in lentiviral or retroviral...pol II promoters allows for more flexibility in expression levels and tissue specificity. Using your shRNA...

  17. Single-cell tracking of lineage and identity with CellTag

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    Blog Post
    ...), now allows scientists to identify the gene expression patterns unique to the few cells that successfully...Unfortunately scRNA-seq only captures the gene expression of cells at a single time point, making it difficult...difficult to investigate how early expression patterns influence reprogramming outcomes. CellTag from ...single-cell level, thus linking ancestral gene expression patterns with reprogramming outcomes (Biddy et...CellTags were detected over a 10-week period. Their expression decreased over time but they were not silenced...MEFs were reprogrammed into iEPs by retroviral expression of transcription factors Foxa1 and Hnf4α, a process...lineage trees and comparing them with the gene expression changes of more than 100,000 cells undergoing...

  18. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as...as a proxy for the endogenous gene – its expression levels (high/low) and patterns (tissue/timing) can...that the gene which GFP was monitoring was being expressed in the same pattern. These systems suggest the...function of a gene. For example, they eliminate the expression of the gene (by CRISPR knockout, shRNA depletion...Tissue-specific genes – the genes of interest may not be expressed in the tissue/cell line you are working with. ...the gene as a result but will still allow for expression pattern monitoring. Enhancer traps Like promoter...trapping of endogenous enhancers to stimulate the expression of the reporter. The system is made up of a reporter...

  19. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight...overnight reaction. STAgR is highly customizable as vector backbones, and gRNA structure, numbers, and promoters...Read the paper in Science Find the anti-CRISPR expression plasmids CRISPR-based method for cell lineage...

  20. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...cells to express Cas9. In these particular experiments, we generated A549 cells stably expressing Cas9 via...wildly variable Cas9 expression (Figure 1). This is a problem because Cas9 expression affects growth rate...variable Cas9-expression was across the different cells (Figure 3).   Figure 2: Cas9 expression in monoclonal...Checking Cas9 expression Overview: By Western blotting (Figure 2, pool), I knew the cells expressed Cas9, but...thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low (even in...against GFP expression, but I didn’t. 2. In the original publication they see GFP expression in about 20%... via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then...
Showing: 181 - 200 of 407 results