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We narrowed to 199 results for: guide rna expression plasmid

Showing: 161 - 180 of 199 results

  1. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ... great collection of ready-to-use rAbs and rAb plasmids!). Since recombinant antibodies conveniently allow... allow for unlimited production, reliable expression, and easy distribution as DNA (Trimmer, 2020), you...antibodies can be produced through a single or dual-plasmid transfection, depending on whether the heavy and...and light chain genes are contained to one plasmid or if they’ve been separated onto two. You’ll first...first transfect your cells with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect ...on the Addgene blog Introduction to Antibodies Plasmid-based Recombinant Antibodies Polyclonal Antibodies...Antibody Purification Protocol Addgene's Antibody Guide References Trimmer JS. Recombinant Antibodies in...

  2. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...It can be very time consuming to design 5 guide RNAs (gRNAs) targeting each of the 1000 genes you’d like...Differentially expressed genes  As the costs for global gene expression analyses like RNAseq are getting ...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...generation sequencing to identify which gRNAs are differentially expressed in the separate groups and, as a ...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...

  3. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...transgene expression. Fluorescent proteins are widely used for tracing and their expression is often made... the mammalian brain (Wickersham et al., 2007; plasmids at Addgene). AAV vectors: A platform for delivering...proteins. For a brief overview see Addgene’s AAV guide. AAV particles are relatively small (20 nm) in size...Cre-dependent vectors are used in transgenic animals that express Cre in only specific cell types, for example only...labelling the subset of infected cells that also express Cre. Another advantage of AAV vectors is that there...their target tissues (adapted from Addgene's AAV guide)  Commonly used serotypes include AAV1 – 9, with...et al. 2014 traced neurons with an AAV9 vector expressing a Cre-dependent fluorescent marker. Injection...

  4. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    Published
    April 2, 2019, 1:44 p.m.
    ... 1998). Interested in plasmids that contain the groucho gene? Check out plasmids deposited by the Shivdasani... several plasmids that contain SHH, you can find them all here! The most popular SHH plasmid however was...spaghetti. We have two plasmids in our inventory from the lab of Eric Wieschaus that express sqh.   cookie monster... to see for yourself. We have one plasmid in our repository, plasmid 39624, that contains cmr.   tinman...website for plasmids that contain tinman or the mammalian homolog Nkx2.5. We also have one plasmid, Addgene...Addgene blog Read our Quick Guide to Working with Drosophila Read our Plasmids 101 blog series Resources...Resources on Addgene.org Find plasmids for your research Deposit plasmids at Addgene  ...

  5. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...the ability to distribute plasmids with the piggyBac™ transposon. These plasmids, when combined with a source... minimize expression levels of your gene of interest, as needed. Controlling the expression levels of ...transfecting with PBx mRNA can boost efficiency over transfecting with plasmids in some cell lines, most...useful resource. While Addgene cannot distribute plasmids with the piggyBac™ transposase itself, please ...genome fully intact, while, with a standard DNA plasmid transfection, a double-stranded break may occur...may be particularly useful when the gene being expressed has threshold effects, such as with signaling ... can also mitigate the deleterious effects of expression of certain genes, such as those that have negative...

  6. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...Endogenous biotin ligases Expression system Stable cell line expressing 1) FLAG-biotin-tagged dCas9,...When these three components are co-expressed, the gRNA targets the dCas9 to the loci of interest and dCas9...lines will have varying levels of dCas9 and gRNA expression. Additionally, CAPTURE 3C-based sequencing ...requires the creation of a stable cell line that co-expresses three components: dCas9 with an added biotin ...that is recognized by biotin ligases normally expressed in eukaryotic cells. Using this tag reduces the...CAPTURE 2.0 to be used with primary cells. Find plasmids for CAPTURE 2.0! When compared to the original...conformation data is compared with changes in gene expression, chromatin accessibility, and epigenetic modifications...

  7. 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...Check out these these plasmids Catch Up on Your CRISPR Background with Our Guide Pages Other Links Check... Lieber A (2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9 Toxicity...CRISPR   Resources on Addgene.org Find CRISPR Plasmids for Your Research Interested in light-inducible...recognition site (Harrington et al., 2017). Inhibit crRNA loadingExample: AcrIIC2 interactions with Cas9 interferes...interferes and prevents the correct assembly of the crRNA-Cas complex (Zhu et al., 2019). Inhibiting DNA cleavage...CRISPR activity in many ways such as inhibiting crRNA loading, inhibiting DNA binding, or inhibiting DNA...

  8. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...dystrophy. While mini-dystrophin expression levels were lower than expression of wild-type dystrophin in normal...Drawbacks: Transgene expression varies with this approach, ranging from ~10% expression in primary human ... an AAV transfer plasmid Resources on Addgene.org: Check out the Addgene AAV guide Find ready-to-use...this AAV successfully mediated transduction and expression of the full-length transgene in vitro and in ...genomes complement each other to restore the full-expression cassette. AAV’s natural ability to reassemble...overlapping split vector has equal or higher transgene expression when compared to using a single over-sized vector...could have a big impact on the rate of transgene expression with an overlapping AAV since both of these variables...

  9. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...out Our CRISPR Guide Pages Find CRISPR Plasmids for Your Research Find Validated gRNAs for Your Research...for future preclinical and clinical work. Find Plasmids for Disease Research on Our Special Collections...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...et al. show that the percentage of dystrophin-expressing muscle cells increases over time, and all three...three groups confirm dystrophin expression via Western blot, at <10% of the wild-type level. Muscle histology... reduce off-target editing. Short-term CRISPR expression would be especially desirable, as it would reduce...

  10. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ...in analysis Plasmid-based proteins: plasmid sequence not accurate Check plasmid sequence for frameshifts...important for highly expressed proteins, including “housekeeping” or ubiquitously expressed proteins used for...protein expression is preferable, and more accurate, than normalizing to ubiquitously expressed proteins...lightly marked on the membrane (1) using the gel as a guide. The gel is then removed from the membrane (2) and... for normalization. Many times the expression levels of these proteins are out of the measurable range...dilutions are in the dynamic linear range (i.e., expression decreases as protein concentration decreases)... expert Review protocol videos for a visual guide to good technique High background Antibody...

  11. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...Unpublished or pre-publication plasmids": Unpublished plasmids or plasmids that will not be published are...and entering a plasmid name,plasmid type, and plasmid description/experimental purpose (right). On the...with your plasmids, and adding all the plasmids at once increases the likelihood that the plasmids are given...easy for you to share plasmids. To achieve this goal, we will archive any plasmids you've deposited with...post as a step-by-step guide to the online deposit process. To deposit plasmids online, you will need ...Spreadsheet". Start your plasmid deposit There are two options to begin your plasmid deposit. Figure 2: Choose... Choose to submit plasmid(s) online or by using a spreadsheet. The plasmid information can be entered ...

  12. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ...often employed to establish cell lines that stably express a gene of interest. While the general approach ...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...compete with host cells for nutrients and can alter expression of receptors, ion channels, and growth factors...after transfecting with the transfer and packaging plasmids. A benefit of the multiple harvest strategy is...on a variety of factors such as the transgene expressed, the viral storage buffers used, and other experimental...will be able to overcome differences in receptor expression. It's all in the mix Even before the lentiviral...infection significantly alters microarray gene expression profiles.” Biotechniques 35.4 (2003). 812-4. ...

  13. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    Published
    April 18, 2024, 1:15 p.m.
    ... (A) Cumulative CRISPR plasmids shipped by year. (B) Cumulative CRISPR plasmids deposited with Addgene...,000 CRISPR plasmids in our repository, and we have shared almost 300,000 CRISPR plasmids and 12,000 CRISPR...these CRISPR plasmids (and libraries)? We wanted to know, so we classified CRISPR plasmids containing a...CRISPR plasmids requested each year by year of deposit (Figure 3), we saw that once CRISPR plasmids become... CRISPR plasmids seem to never truly go out of style. Instead, an almost even mix of plasmids (by age)...Total CRISPR plasmids deposited by country; square size is proportional to number of plasmids deposited ...Total CRISPR plasmids requested by country; square size is proportional to the number of plasmids requested...

  14. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    Published
    July 20, 2021, 1:15 p.m.
    ...level and timing of expression can also be affected by the tag.  Protein expression, measured using antibody...Addgene.org Visit the Fluorescent Protein Guide Find antibody plasmids for your experiments ...degree of internal complexity such as extensive membranes produces a greater SSC.  Protein expression, measured...Have you ever wanted to measure expression of your protein of interest in a single cell? Or perhaps, ...applications such as measuring target protein expression levels, assessing post-translational modifications...system (Figure 1).   Figure 1: Cells expressing a particular surface receptor are stained with...interrogated with a fluorescent light source, a cell expressing the associated fluorophore or stained with the...

  15. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    Published
    Dec. 13, 2018, 1:14 p.m.
    ...production. Whether redesigning viral packaging plasmids, investigating different viral production strategies... of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...resources of the Addgene blog Read our beginner's guide to viral vectors Find all of our viral vector blog...Rather than being released into the cell culture supernatant, the viral particles associate with the cell ...production phase, improves adenovirus production." Journal of biotechnology 157.1 (2012): 228-236. PubMed ... trafficking of adeno-associated virus/phage." Journal of Biological Chemistry (2012): jbc-M112. PubMed...

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

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ... in a plasmid with loxP sites such that their expression is turned on or off in cells expressing Cre. ...Addgene.org Check out Our Fluorescent Protein Guide Pages Find Plasmids for Subcellular Localization Find Fluorescent...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...efforts have been made to create transgenic mice expressing fluorescent protein reporter constructs in the...above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after...Introduced to immunology by three papers in the journal Science in 2002. Now, this technique and the in...

  17. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 3:42 p.m.
    ... Hub AAV Guide AAV Packaged On Request Browse all in-stock AAV preps Browse all AAV plasmids ...viral capsid, but specificity doesn’t end there: expression from rAAVs also depends on your choice of promoter..., et al., 2006). AAV2 capsids are available as plasmids in the repository and can be selected as a serotype...efficiency in multiple tissues, rapid transgene expression, and can cross blood vessel barriers. It is especially..., et al., 2006). AAV8 capsids are available as plasmids in the repository, as part of our in-stock viral...retinal cells (Issa et al., 2023). AAV9 packaging plasmids are available at Addgene. AAV9 vectors are not...the axon to the cell body and driving transgene expression (Tervo et al., 2016). This functionality makes...

  18. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...Addgene.org Fluorescent Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors Keima, another...monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 localizes to the cytosol in an inactive...yeast Rosella is targeted to the cytosol when expressed without a signal sequence and to the mitochondria...measuring the amount of radioactivity in the culture supernatant (this is indicative of protein degradation). ...

  19. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    Published
    May 4, 2023, 1 p.m.
    ...Additional resources on Addgene.org CRISPR Guide CRISPR Plasmids and Resources CRISPR History and Development...generation of multiple edits or implementation of expression systems to accurately mimic a disease phenotype...best served by this type of targeted design. Alternatively, a disease model can be generated, and drugs...

  20. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...Resources ATCC Animal Cell Culture Guide Thermofisher Guidelines for Maintaining Cultured Cells Invitrogen...Invitrogen Cell Culture Basics Guide Abcam Cell Culture Guidelines Resources on the Addgene Blog Learn ...along with Down-Regulation of HIF-1α by NDRG2 Expression Attenuates Proliferation and Invasion in Renal...Protein Localization Resources on Addgene.org Find Plasmids for Mammalian Cells ...identification of glioma-initiating cells. The International Cell Line Authentication Committee includes ...cell culture outlined in this article. Now many journals strictly require evidence of cell line provenance...
Showing: 161 - 180 of 199 results