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We narrowed to 276 results for: guide rna expression plasmid

Showing: 201 - 220 of 276 results

  1. Antibodies 101: Producing Recombinant Antibodies

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    Blog Post
    ... 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
    ...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

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    Blog Post
    ...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

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    Blog Post
    ... 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. Lentiviral Prep Service

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    Collection
    ...ways. Cas9 proteins can be guided to specific genomic sites by an RNA guide (gRNA) that harbors targeting... a list of gRNA plasmids targeting specific genes. For more information on CRISPR plasmids and function...are made from plasmids that are currently in our repository. You can obtain the plasmid from us or receive...biological processes. For more information, see our Guide to Plasmid Pooled Libraries . ID Name Description Genes... of available viruses based on plasmids in our repository that express fluorescent proteins. ID Name Insert...function, see our CRISPR plasmids resource page . Cas9 for Editing Genomic Sequences To perform genome edits...52962 lentiCas9-Blast Sp Cas9 Cut Blasticidin Expresses human codon-optimized S. pyogenes Cas9 protein...

  6. CRISPR Plasmids - gRNAs

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    Collection
    ...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...using any of these gRNA plasmids and review the publication associated with each plasmid for more information...information on the experimental use of these plasmids. ID Plasmid Gene/Insert Vector Type PI Publication Do...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail... CRISPR gRNAs CRISPR Plasmids: gRNAs Browse CRISPR Plasmids...CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The table below lists gRNA plasmids...gRNA expression vectors available from Addgene. CRISPR...

  7. PiggyBac-ing Through the Genome Editing Field

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    Blog Post
    ...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...

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

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    Blog Post
    ...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...

  9. CRISPR Plasmids - Double-Strand Break (Cut)

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    Collection
    ...Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize...below for CRISPR plasmids designed to introduce a DSB. Plasmids are available for expression in mammalian ...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Drosophila ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Plant ID Plasmid Gene/Insert... elegans ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast ID Plasmid Gene/Insert...Zebrafish ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Xenopus ID Plasmid Gene/Insert...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...

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

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    Blog Post
    ...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...

  11. AAV Production in HEK293 Cells

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    Protocol
    ...must be optimized. Plasmids for transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene of interest...of each plasmid needed to have a 1:1:1 molar ratio with 2 mg total DNA per CS5 Plasmid Plasmid Size (bp...molar ratio of each plasmid: 2000 μg / 24,961 bp = 0.08 μg/bp Therefore, for each plasmid we need: Sample ...may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference Introduction... 1.00 727.6 pHelper 11,854 1.00 1,185 Transfer Plasmid 5,842 1.00 584.2 Total bp 24,961 In total, we would... the total number of base pairs from all three plasmids, we can determine the total μg/bp we need to achieve...Needed: 949.8 μg / 1.0 μg/μL = 949.8 μL Transfer Plasmid: 0.08 μg/bp × 5,842 bp = 468.1 μg Volume Needed...

  12. CRISPR Plasmids - Empty gRNA Vectors

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    ... on the plasmid. gRNA plasmids that do not co-express a Cas protein require a separate plasmid that does...constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 ...for sgRNA expression. Chen pCFD4-U6:1_U6:3tandemgRNAs Drosophila Single plasmid for the expression of ...Vectors CRISPR Plasmids: Empty gRNA Expression Vectors Browse CRISPR Plasmids By Function Genome Editing Cut...can be used to express multiple gRNAs from a single plasmid with or without expression of Cas9. Highlighted.... Can express Cas9 with up to 3 gRNAs. Frew pSQT1313 Mammalian Single plasmid for the expression of multipe...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...

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

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    Blog Post
    ...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...

  14. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...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...

  15. Troubleshooting and Optimizing a Western Blot

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    Blog Post
    ...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...

  16. How to Deposit Your Plasmids with Addgene

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    Blog Post
    ...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 ...

  17. Getting the Most from Your Lentiviral Transduction

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    Blog Post
    ...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. ...

  18. Twenty Years of Addgene Sharing: CRISPR

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    Blog Post
    ... (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...

  19. Antibodies 101: Flow Cytometry

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    Blog Post
    ...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...

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

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    Blog Post
    ...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...
Showing: 201 - 220 of 276 results