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We narrowed to 407 results for: expression vector

Showing: 241 - 260 of 407 results

  1. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ...protein of interest into a vector encoding a fluorescent tag. Once expressed as a fluorescent protein fusion...bacterial components. For example, the Rainbow Vectors from the Mariette Barbier Lab can be used to fluorescently...Barbier, Mariette, and F. Heath Damron. "Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling...subcellular structure or protein complex. By coexpressing both the marker protein and the protein of interest...

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

    Type
    Blog Post
    ...promoter is expressed. Cre-mediated deletion of one or several of these genes permits the expression of genes...activate the expression of a different fluorescent protein. Therefore, each cell could express a unique combination...copy of the transgene expressing either YFP or CFP, and cells that do not express Cre fluorescing with ...the low expression of XFP of Brainbow by incorporating a Tet-Off system to boost XFP expression. The increased...variety of colors when a combination of XFP are expressed. Each cell remains the same color for its entire...of the downstream OFP, GFP, or RFP. Cells that express the construct but do not undergo recombination ...truncated YFP. In Thy1-Brainbow mice, some neurons expressed as many as 16 copies of the injected transgene...

  3. Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight

    Type
    Blog Post
    ... channels, such as ChR2 and NpHR, can even be expressed simultaneously and still be individually activated... like spontaneous swimming. To do so, the lab expressed the LiGluR opsin in different subsets of spinal...optogenetics blog posts Read blog posts about viral vectors Subscribe to the Addgene blog Resources on Addgene.org...

  4. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    ...several projects and this is one related to gene expression. One goal of the ENCODE project is to look at...assays, not just transcription factors. The second vector is a homology directed repair donor plasmid with...flexible. Other investigators can easily adapt these vectors for use with their favorite selection cassette....

  5. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...recombination, RFP is expressed. Cre can mediate one of two deletions to allow YFP or M-CFP expression; these deletions...editing may be the hot new way to manipulate gene expression, but other gene manipulation systems remain valuable...ways to spatially and temporally control gene expression, especially in in vivo models, and new Cre-lox...fluorescent protein in the construct, OFP (orange), is expressed when Cre is absent; stochastic Cre excision results... and inversion. The Brainbow-2.1 construct can express one of four colors (n-GFP, RFP, YFP or M-CFP.) ...remaining cassette can invert as long as Cre remains expressed; once Cre is removed from the system, the fluorescent...coding sequence is closest to the promoter will be expressed. This system differs from Brainbow-1.0 in that...

  6. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...determined by comparing cells expressing only eGFP expression to cells expressing eGFP and mCherry. ACE was...mCherry and a T2A site and expressed in cells. mCherry is constitutively expressed to identify successfully...that can be repaired by NHEJ restoring mCherry expression. Cells can then be sorted using Fluorescence-...BFP variant, a BE4am-Cas9 base editor, and sgRNA vectors for the BFP-to-GFP conversion and to target specific...integrated it into human and mouse cells that expressed either Cas9 or an optimized cytidine base editor... of a mutated start codon to initiate protein expression, it can be used for an array of detectable proteins...

  7. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...drive gRNA expression is dependent upon the cell type in question. In plant cells, gRNA expression is achieved...Cas9 expression is commonly driven by plant-derived RNA pol II promoters which regulate expression of longer...mRNAs for gene expression). Examples of commonly used RNA pol II promoters for Cas9 expression include the...system in plant cells. Like other model systems, expression of S. pyogenes Cas9 or Cas9 variants (hereafter...OsU6 or OsU3, which are commonly used to drive expression of small RNAs in their respective species. Addgene...with other model systems, multiple gRNAs can be expressed to modify several genomic loci at once (get more...the ubiquitously expressing cauliflower mosaic virus 35S promoter (CaMV 35S) or ubiquitin promoters (Belhaj...

  8. Multifunctional Neural Fibers: Modulation from Gut to Brain

    Type
    Blog Post
    .... They started by delivering an AAV construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive...potential afterwards. This measurement trended with expression of the opsin in the target neurons, validating...studies Channel rhodopsins for optogenetics Viral Vectors 101: Chemogenetics ...

  10. Developing Lab Management Software for Biology

    Type
    Blog Post
    ...collisions you may encounter: vector, sequence, insert, trait, factor, expression, etc.There are also going...terms (plasmid, 5-prime, CRISPR, growth strains, vector backbone, etc.) and you simply don't have enough...

  11. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...plasmid contains humanized Cas9 expression cassette and sgRNA expression cassette. The target sgRNA can...a) Scheme of validation for DSB mediated EGFP expression cassette reconstitution. When the target sequence...annealing) took place and reconstituted the EGFP expression cassette. (b) pCAG-EGxxFP target plasmid and ... PX330 plasmids (or similar plasmids for gRNA expression), one gRNA at a time. After 48 hours of incubation...homology directed repair, and resulting in the expression of EGFP. Using this technique, you can assess...clone the PCR products into a pBluescript cloning vector and sequence the resulting plasmid. It can take...mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA." Scientific reports...

  12. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...This tracrRNA was combined with the sgRNA expression vector allowing the screen to be conducted with only... activation domain (CRISPRa) to regulate gene expression. Some CRISPR libraries contain many sgRNAs per... sgRNA library In addition to inhibiting gene expression, dCas9 can also be used for transcriptional activation...screens can help reveal the function of lowly expressed genes or genes that are more drastically affected...protein-tagging system for signal amplification in gene expression and fluorescence imaging." Cell 159.3 (2014):...screens performed with open reading frame (ORF) overexpression libraries for resistance to MEK inhibition....inhibition. Although both Calabrese and the ORF overexpression library identified several overlapping genes, most...

  14. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ... al., 2016). Using lentiviral vectors from Addgene, the lab will express these candidate genes in TS iPSCs... It’s possible that solute channels typically expressed on other cell types can compensate for or bypass...defects. Eight genes from the X chromosome are expressed at significantly reduced levels in women with ...

  15. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...subset of RSMs that regulate gene expression) that uniquely express selectable and/or sortable reporters...programs that specify which genes should be expressed or not expressed in each state. To help researchers adapt...register”. Each chemical input to an RSM drives expression of a distinct large serine recombinase that recognizes...system), these reactions are irreversible. Once expression of a particular recombinase is triggered, its...possible downstream states that can be achieved by expression of other recombinases. Multiple orthogonal attB-attP...present a two-input, five-state GRSM register that expresses a unique combination of red, blue, and green fluorescent...state detection with flow cytometry. However, expressing reporters will consume cellular resources and...

  16. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...activate, we would recommend using a cDNA overexpression vector rather than going through all the troubleshooting...used in therapeutics, as it can increase gene expression at certain loci to create changes in phenotypes...: once the environmental trigger leaves, gene expression is no longer activated. However, there are several...up-regulated. For instance, generally, the more highly expressed a gene is under native conditions, the less activation...libraries, which consist of plasmids that over-express coding sequences from a given cell type or organism... for altering DNA sequence or regulating gene expression.     References Behan FM, Iorio F, Picco G, Gonçalves...Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging. Cell 159:635–646 . ...

  17. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...is then digested and ligated into an appropriate vector. Gibson Assembly, GeneArt Cloning, and SLIC can...of recombineering. In recombineering, bacteria express phage recombination machinery, which facilitates...products are directly transformed into bacteria expressing recombineering proteins Red/ET from a temperature-sensitive...

  19. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...gene drive is also the easiest: don't use a DNA vector that encodes both Cas9 and sgRNA. What if I want...spread exclusively through populations that already express Cas9 from an unlinked locus, while a Cas9+sgRNA...attempting to build gene drives in tropical mosquito vectors of diseases such as malaria and dengue are currently...

  20. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Cells You may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference...transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene of interest Triton X-100 Benzonase/...
Showing: 241 - 260 of 407 results