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We narrowed to 369 results for: Cre expression vector

Showing: 241 - 260 of 369 results

  1. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...in therapeutics, as it can increase gene expression at certain loci to create changes in phenotypes. For...activate, we would recommend using a cDNA overexpression vector rather than going through all the troubleshooting...: 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 . ...

  2. 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...

  3. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...loxP sites such that their expression is turned on or off in cells expressing Cre. The result of a system...15 years efforts have been made to create transgenic mice expressing fluorescent protein reporter constructs...allows the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell...immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes...above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after...useful tool for marking and tracking cells is the the Cre/lox recombination system. A researcher can flank ...respectively. The permanent labelling of cells using cre-lox is advantageous when compared to photoactivable...

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

    Type
    Blog Post
    ...avoid accidentally creating a gene drive is also the easiest: don't use a DNA vector that encodes both ...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...Cas9 should be aware of the potential hazards of creating “gene drives” capable of spreading through wild...does making transgenic organisms with Cas9 risk creating a gene drive? The simplest way to make a gene ...appropriate flanking homology into a germline cell can create an RNA-guided gene drive. In principle, this event...

  5. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to ... closed RNAs seem to be conserved in mammals, expressed in neurons, and are especially abundant in neuronal...resulted in a cell migration defect. They have created a circRNA website so scientists can view public...developments. Using the Tet-on system, his lab has created a tetracycline cas9 inducible knock-out system....

  6. 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...harnesses the power of bacterial recombineering to create insertions, deletions, and substitutions - at the...

  7. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...stability. Additionally, the choice of vector can impact the expression level of an antibody (Ayyar et al....such as parasites or microbes, or to proteins expressed on the surface of cells that have been infected...the protein out of solution, or isolate cells expressing this protein. Read on to learn more about antibodies...context of the immune system, each isotype is expressed at different times of the immune response and ...qualitative and quantitative measurement of protein expression in cell lysates, whole cells, or tissue samples...frequently used to compare relative levels of protein expression between cell types or treatment conditions. They....    Figure 4: Western blot showing GFP expression in cells that have been transfected with a plasmid...

  8. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...consist of a group of genes/proteins that, when expressed, normally encourage cell survival, growth and ...Clone Collection 2.0. The collection is a set of vectors for use with the Gateway Cloning Platform (Life... is that it enables the simple construction of vectors to study the function of any of these genes through...as 30% of all cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations...he or she can use to study it (as seen in the screenshot below). Currently, the page includes interactive... Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer Library Can Be Used...

  9. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...the phage display screens are subcloned (using FX cloning) into the expression vector pSb-Init, which attaches...their small size, nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target...sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins...to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure 1: ...threonines at positions that can be randomized for creating the libraries (Zimmermann et al., 2018). Find ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...the candidate sybodies in their coat protein to screen for binding to the target protein. The protocol...

  10. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...flow cytometry, and mRNA expression.   If your protein of interest is expressed at similar levels in ...useful to produce a sample with mixed expression, with the expression measurable through an epitope tag such... sets, you can increase confidence that the labeling observed represents the expression level of the target...epitopes. Detection of overexpressed (tagged or untagged) protein When expression plasmids are available...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...caused by overexpression. Additionally, if the cell line used has endogenous levels of expression, this can...preferred. The overexpression approach is therefore best suited to cell lines without any expression of the target...

  11. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...that the percentage of dystrophin-expressing muscle cells increases over time, and all three groups confirm...myoblasts; when injected into DMD mice, these cells expressed functional dystrophin. Editing genes in vivo is..., the CRISPR components would only need to be expressed for a short period of time, which would prevent...confirm dystrophin expression via Western blot, at <10% of the wild-type level. Muscle histology is improved...stem cells 2. Remove any immunogenicity of the AAV vector Once delivery has been optimized, it’s important... reduce off-target editing. Short-term CRISPR expression would be especially desirable, as it would reduce... genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy. Science...

  12. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...marvel of DNA compaction, an enhancer can increase the expression of a gene that is millions of base pairs...genome math expression for NANOG enhancers, we’re ready for step 2: design a CRISPR screen to disrupt ...CRISPR screen. We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an...Putting it all together, we get the following expression for step 1: (H3K4me1 + H3K27ac) – H3K4me3 The...stem cells, and conclude with a targeted CRISPR screen that can disrupt these enhancers. A real-world ...that have NANOG binding sites 2. Design a CRISPR screen to target and disrupt the NANOG enhancers 3. CRISPR...you’re interested in designing your own CRISPR screens for enhancers, check out the Appendix. Conclusion...

  13. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...deliver in low-capacity vectors, such as adeno-associated viral (AAV) vectors. In 2017, Zetsche et al....multiplexing approach using a single crRNA array to express up to 4 crRNAs. Zetsche et al. also suggest that...PAM site, Cpf1 cleaves in a staggered fashion, creating a 5 nucleotide 5’ overhang 18-23 bases away from...mediated editing, as Kleinstiver et al. saw no decrease in Cpf1 activity with 4-6 base deletions at the...

  14. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...al., 2024). In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study...than SpyCas9, allowing them to be packaged in AAV vectors, potentially with multiplexed targets.   ...showed that AAV-delivered CHARM could silence the expression of prion protein in mouse brains for at least...gene product; it simply turns the target protein expression off. Plus, DNA methylation is a long-lasting ...-silencing CHARM variants that stop their own expression after silencing their target — potentially improving...by targeting marker proteins that are uniquely expressed in a given cell type. For example, glial fibrillary...GFAP) is a glia-specific intermediate filament expressed in astrocytes throughout the brain, making it ...

  15. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ... plasmids expressing Cre recombinase and filter the results by expression system and vector type. Find...or control gene expression or skip ahead to browse highlighted plasmids expressing Cre , Flp , Dre , other...Summary of Cre, Dre, and Flp Recognition Sites. Controlling Recombinase Expression By expressing the enzyme...interest will prevent gene expression in the absence of Cre. In the presence of Cre, the stop codon is excised...lox-STOP-lox shRNA constructs, Cre expression promotes shRNA expression. Gene Switch: These constructs...Genes A and B. When Cre is absent, only Gene A is translated correctly. Cre expression excises Gene A and...double-crossover event. Toxicity Cre-lox recombination is a very useful tool, but Cre expression can be toxic in some...

  16. Brain Initiative Collection

    Type
    Collection
    ...CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA AAV vector for Cre expression in forebrain GABAergic interneurons...-AAV5 pAAV-CAG-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...AAV8 pAAV-Ef1a-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...AAV9 pAAV-Ef1a-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...combination with Cre-dependent transgenic mouse lines or co-infection with Cre dependent viral vectors. Jonathan...AAV-mediated expression of voltage sensor under the Syn promoter, Cre-dependent expression; soma localization...

  17. Zhang Lab CRISPR Page

    Type
    Collection
    ...2015). The dual vector system uses S. pyogenes Cas9 (SpCas9), using one vector to express SpCas9, and another...delivering Cre and sgRNA to a Cre-dependent mouse or sgRNA to a constitutively Cas9-expressing mouse. Described...luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone with luciferase-2A-Cre recombinase Vectors for brain...mouse toolbox protocol 637.8 KB CRISPR-Cas9 Cre expression vectors for cancer modeling Using Cas9 mice, Platt...pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR This plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase...target site sequence (20bp). CRISPR-Cas9 Cre expression vectors for genome editing in the brain Using Cas9...

  18. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ... lentiviral packaging systems, and luciferase expression. On individual material pages, we added curation...curation fields for purpose, promoters, expression types, and more, all of which help a researcher understand... Inducible Plasmids Empty Backbones Bacterial Expression Systems AAV Plasmids Blogs Origin of ... CRISPRTwenty Years of Sharing: Addgene's Viral Vector Service...ve learned that increased accessibility is achieved through interconnected incremental changes. It was...When Addgene opened in 2004, we focused on creating a repository that made it easy to share plasmids,...identified and offered a number of solutions that increased accessibility, such as using couriers that refill...

  19. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...for quantitative imaging, Ming Hammond’s group created Spinach2 using site-directed mutagenesis of Spinach...for in vivo studies. Dimerization of Broccoli increases its fluorescence by about 1.8-fold, and may be...for greater contact with its RNA aptamer. This increased interaction between Corn and its small molecule... to TO1-biotin. Two of these variants exhibit increased brightness compared to Mango, and all are photostable...

  20. Kazuhiro Oka Lentiviral Vectors

    Type
    Collection
    ...Collections Gene Vector Core Viral Vectors Gene Vector Core Viral Vectors Lentiviral Transfer Vectors Adenoviral... transfer vectors and generating new viral vectors. Download the Lentivirus Transfer Vector Production...Lentiviral Vector ID Purpose pCDH-EF1-DIO-copGFP 72253 Expresses copGFP under EF-1 promoter when Cre is expressed...by another vector pCDH-EF1-DIO-tdTomato 72254 Expresses tdTomato under EF-1 promoter when Cre is expressed...promoter when FLP is expressed by another vector pCDH-EF1-Fon-tdTomato 72261 Expresses tdTomato from the ... promoter when FLP is expressed by another vector pCDH-EF1-FLPe 72262 Expresses FLPe from the EF-1 promoter...lentiviral and adenoviral vectors read our Lentiviral Vector Guide and Adenoviral Vector Guide Find more plasmids...
Showing: 241 - 260 of 369 results