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We narrowed to 294 results for: expression vector

Showing: 181 - 200 of 294 results

  1. 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 ...

  2. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ...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 ...

  3. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ... 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...

  4. 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...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...

  6. Five Popular Model Organisms

    Type
    Blog Post
    Published
    April 11, 2019, 12:39 p.m.
    ...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...

  7. 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 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...

  8. 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...

  9. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...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 ...

  10. 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...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...

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

    Type
    Blog Post
    Published
    May 9, 2019, 1:15 p.m.
    ...), 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...

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

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...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...

  14. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...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...

  15. Antibodies 101: Affinity Tags

    Type
    Blog Post
    Published
    Sept. 19, 2023, 1 p.m.
    ... single protein is expressed, with the intent of purification, it will be expressed along with all other...other cellular proteins within the expression system (bacteria, yeast, mammalian, etc.). To individually...about that protein that no other protein in the expression system has. This is where affinity tags come ...peptides which do not naturally exist in most expression systems that can be added to the N or C terminus...on: The protein you are trying to purify The expression system If you want the tag to remain on your ...MBP tag, although large, can actually increase expression and solubility of the tagged protein (Fox and...Ready to start tagging? We are here to help with vectors, protocols, antibodies, and more!   Resources ...

  16. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ... information on viral vectors, including transduction download our Viral Vectors 101 eBook. Bacterial ...bacterial or eukaryotic cells via a virus or viral vector. One example are bacteriophages that attach to ...scientists commonly use phagemids, a DNA cloning vector that contains both bacteriophage and plasmid properties...transient cell lines, where a gene of interest is expressed but not integrated into the genome and stable ...

  17. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    Published
    May 28, 2024, 1:45 p.m.
    ...transgenic protein for the HEK 293T cells to express: CD45. Upon expression, CD45 is incorporated into the plasma...CD45, but is separately expressed. (The reason for having a separately expressed transduction reporter ...manage to express CD45. Thus, if transduction efficiency is much higher than CD45 expression, this could... example, point at issues with the double-expression vector.) In our flow experiment, we'll want to identify...CD45+ cells (CD45 expression) and assess our transduction efficiency (eGFP expression).     The first ...detected through a cell surface label. The viral vector also encodes for eGFP, which serves as a transduction...rest of our panel. Lastly, we want to detect the expression of our protein of interest: CD45. I am using ...

  18. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...genome editing, activating gene expression, repressing gene expression, and much more. But there’s one... T, Lieber A (2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9... means they can be delivered in vivo using AAV vectors. Having a way to shut off CRISPR-Cas systems prevents...

  19. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...screens Differentially expressed genes  As the costs for global gene expression analyses like RNAseq are...test which of the differentially expressed genes found in a gene expression dataset are causal for a phenotype...differentially expressed genes that are known drug targets, or all differentially expressed genes that code...sequencing to identify which gRNAs are differentially expressed in the separate groups and, as a consequence, ...CRISPR screens designed to target differentially expressed genes can be the next stage in making sense of...interactions to a large extent involve receptors expressed on the cell surface. Thus, a screen can be used...The gRNA oligos are subsequently cloned into your vector of choice using the adapter sequences that you ...

  20. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    Published
    Nov. 8, 2018, 3:47 p.m.
    ...genes is from Proteobacteria to mammalian expression vectors. And more broadly, their data revealed that...a distantly related organism, the gene may not express well because of these different codon usage biases...closely resembles that of the host organism, it may express protein more efficiently. To classify gene sequences...rare codons, sometimes intentionally modulating expression in ways we don’t quite understand yet,” Kunjapur...synthetic. They also found out that the most common expression system within the Addgene repository is mammalian...
Showing: 181 - 200 of 294 results