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We narrowed to 278 results for: guide rna expression plasmid

Showing: 101 - 120 of 278 results

  1. Joung Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids

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    Collection
    ...Reporter Gene Guide RNA expression plasmids for endogenous human genes Guide RNA expression plasmids for endogenous... CRISPR-Cas/RGN expression plasmids for use in zebrafish Guide RNA expression plasmids for the EGFP Reporter...CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids You may also like... CRISPR Guide CRISPR Protocols gRNA Design...specificities Dimeric CRISPR RNA-guided FokI Nucleases (RFNs) CRISPR-Cas/RGN expression plasmids for use in human ...CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids Genome...consist of two components: a short ~100 nt guide RNA ( gRNA ) that uses 20 variable nts at its 5’ end ... recently described gRNA and Cas9 expression vectors and validated the use of RNAs produced by these vectors...

  2. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ...webpage will have information on where the gene is expressed within the fly, whether stocks of mutant flies...Snapdragon tool helps you select primers to synthesize RNA for gene silencing. Additionally, they have fly collections...of flies for in vivo gene knock down and gene expression. In the next post, you’ll learn the fundamentals...Drosophila CRISPR Resources Deposit Your Drosophila Plasmids ...tools to help you knock down, knock out, and overexpress YFG in a Drosophila cell line. Knock downs in...

  3. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    Blog Post
    ...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...provide synthesized SARS-CoV-2 expression plasmids available for many expression systems: bacterial, yeast,...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...spike protein to improve expression and pseudotyping efficiency. This plasmid was later used by another...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from... labs also deposited plasmids for producing COVID-19 diagnostic enzymes. Plasmids in this toolkit include...

  4. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    Blog Post
    ...vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system enables us to express the CRISPR...enChIP Plasmids? All Hodaka Fujii lab plasmids Sign up to receive email alerts when new plasmids from ...world have requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using... shopping for a wide range of plasmids. The merit of depositing plasmids to Addgene has been spectacular...isolated for identification of associated proteins, RNAs, and other genomic regions. I believe that enChIP...user-friendly and precise. We will deposit the plasmids of these systems to Addgene in the near future...Community Addgene: In 2012 your lab first deposited plasmids with Addgene. In 2014, you joined Addgene's Advisory...

  5. Zhang Lab CRISPR Page

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    Collection
    ...with the guide RNA. For most applications, we recommend using Cas9 with the single guide RNA (sgRNA), which...SpCas9n, D10A nickase) + single guide RNA: These plasmids contain two expression cassettes, a human codon-optimized...SpCas9-2A-EGFP and single guide RNA 62988 : PX459; SpCas9-2A-Puro and single guide RNA 48873 : PX460; SpCas9n...) and single guide RNA 48140 : PX461; SpCas9n-2A-EGFP (D10A nickase) and single guide RNA 62987 : PX462...FACS sorting SaCas9 + single guide RNA: These plasmids contain two expression cassettes, a human codon-optimized...D10A nickase) + CRISPR RNA array + tracrRNA: This plasmid contains three expression cassettes. To target...both the SaCas9 gene and its single guide RNA (sgRNA) into one plasmid (Ran et al., Nature 2015). The dual...

  6. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...genomic site specified by a guide RNA (gRNA) sequence. Users can design gRNAs to induce the break at a specific...co-injection of the targeting vector and a Cas9 expression plasmid will insert the desired genetic tag and marker...as a universal donor plasmid. The CRISPaint toolkit includes several donor plasmids for protein tagging...the first site accepts the sgRNA target sequence for U6 promoter expression and the second site accepts...need 3 vectors:  1) a gRNA vector that targets the gene of interest, 2) a plasmid to specify the reading...allows users to generate a plasmid containing a direct fusion of the desired gRNA to the U6 promoter, and...The Jorgensen Lab recently developed a modular plasmid assembly toolkit called SapTrap to improve CRISPR...

  7. Plasmids 101: Modular Cloning Applications and Kits

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    Blog Post
    ...conditional gene expression, multiplexed CRISPR, and chromosomal integration of larger expression cassettes ...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within...comparisons between CRISPR components expressed from different plasmid backbones. They noted that backbone...or distance in the placement of plasmid elements can impact expression levels and consequently convolute...Cloning (MoClo) Guide Addgene’s eBook Collection More resources on the Addgene blog Plasmids 101: Golden...modifications which can be especially difficult in alternative expression systems such as bacteria. If your yeast... tools can be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even...

  8. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...locus of a bacteria host encodes multiple guide RNAs (gRNAs) on a single array to target the genome of...including previous tRNA-gRNA arrays, that rely on an upstream lengthy promoter to express the gRNA array, DAP ...assembled tRNA-gRNAs is placed downstream of a lengthy promoter, which is used to drive the expression of the... the array. After expression, the individual gRNAs are released by endogenous tRNA processing machineries... Gao Lab demonstrated that the tRNA-gRNA array can drive the expression of itself, without using any additional...design examples of using Addgene plasmids hCtRNA_FT (#186715) and hCtRNA_VT (#186716).   Fig. 2: ...scalable and efficient in plasmid and viral vectors. In applications when repetitive tRNA sequences must be avoided...

  9. Plasmids 101: Repressible Promoters

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    Blog Post
    ...to be expressed. These permit exquisite control of gene expression and tracing of gene expression across... basal expression. Co-expression of repressor QS along with QF leads to intermediate expression from QUAS...transgene permits directed expression of that gene in cells that also express GAL4. Usage of this system...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...basal state, and co-expression of the inducer QF can increase QUAS driven expression by ~3,300-fold in ... Shen. “Controlling gene expression with the Q repressible binary expression system in Caenorhabditis ... with transgene or siRNA expression from the other. The Q system Potter et al. from Liqun Luo’s lab developed...

  10. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

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    Blog Post
    ...and Cas9 expression plasmids. These results are in line with previous research showing that the method of...Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources Learn about viral production...in place of SpCas9. gRNA A gRNA targeting a DNA sequence of interest is expressed in 293T cells and loaded...same guide. Nanoblades also had lower off-target effects when compared to DNA transfection of gRNA and ...infiltrate cells. Options for CRISPR delivery include plasmid transfection, RNP electroporation, and viral transduction...Nanoblades, different viral envelope protein(s) are expressed to pseudotype Nanoblades. The team found that ...types they tested. Interested in using Nanoblades? Plasmids are available from Addgene. Genome editing primary...

  11. Degrading DNA with Cascade-Cas3

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    Blog Post
    ...Liao, M., & Ke, A. (2018). Structure basis for RNA-guided DNA degradation by Cascade and Cas3. Science,...Additional resources on Addgene.org CRISPR guide Cascade-Cas3 Plasmid collection ...its partner in crime, Cascade. Find Cascade-Cas3 plasmids in Addgene's collection! A “complex” system Cas3...challenging. It can be difficult to ensure comparable expression of all components of Cascade for maximum efficiency... Various labs have designed all-in-one Cascade plasmids — such as the Tomoji Mashimo lab and the Tilmann...efficiency of these systems, though consistent expression can still be a challenge. A multitasking Cas ...a cost — the previously mentioned issues with expressing all Cascade components can make Cas9 easier to...

  12. Harnessing Bacterial Toxins for Allelic Exchange

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    Blog Post
    ...Tight control of toxin expression is absolutely necessary, because leaky expression of toxin could select...Central PMCID: PMC4862005. Jiang, Wenyan, et al. "RNA-guided editing of bacterial genomes using CRISPR-Cas...Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria." Plasmid 51.3 (2004): 246-255. PubMed...introduce a mutation in the chromosome is to disrupt expression of a gene with an antibiotic resistance cassette...in the chromosome, sometimes interfering with expression of surrounding genes. However, there are ways...exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate off-target mutations...a mutated allele introduced from a conjugative plasmid replaces the chromosomal copy. This is a versatile...

  13. Western Blot

    Type
    Protocol
    ...may also like... Antibody Plasmid Collection Antibody Blog Posts Antibody Guide Immunocytochemistry Protocol...include a positive control sample that you know expresses the protein, such as cells transfected with the...interest, and a negative control sample that does not express the protein of interest....to support reproducibility in science. General Guidelines Acrylamide percentage varies in SDS-PAGE gels...6 cells for 5 min at 100 x g . Gently remove supernatant. Resuspend cell pellet in 1 mL of 1X PBS and ...Centrifuge for 5 min at 100 x g . Carefully remove supernatant. Resuspend cell pellet in appropriate volume ... for 15 min at 14,000 x g at 4 °C . Transfer supernatant to a clean microcentrifuge tube. Use the lysate...

  14. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

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    Blog Post
    ... [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]. Many of the plasmids used for...four reprogramming factors can be delivered and expressed in multiple somatic cells via various methods....Neurons [32, 33, 34] You can find a variety of plasmids for differentiation here. Transdifferentiation...transdifferentiated in vivo to pancreatic β-like-cells with the expression of pancreatic and duodenal homoeobox gene1 (PDX1...transdifferentiation is usually carried out by expressing transcription factors specific to the lineage.... Davis, R.L., H. Weintraub, and A.B. Lassar, Expression of a single transfected cDNA converts fibroblasts...Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced...

  15. Antibodies 101: Affinity Reagents

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    Blog Post
    ...enough to be expressed in bacterial and other cell systems. Since the majority of these alternative reagents...Antibody GuideRecombinant antibodies available from AddgeneBrowse our recombinant antibody plasmid collection...I love Addgene's new Antibody Guide and not just because I helped create it. But while antibodies, as...could take the needed steps to continue where the guide left off - namely, a blog post devoted to the other... of disulfide bonds, monobodies can be easily expressed in transfected eukaryotic cells, allowing for ...Aptamers are not proteins at all. These high-affinity RNA molecules can bind to a wide variety of targets, ...Resources Additional resources on the Addgene blog Plasmids 101: Aptamer FluorophoresAntibodies 101: scFVs...

  16. GPCRs: How Do They Work and How Do We Study Them?

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    Blog Post
    ...ligand-bound GPCRs of interest. The TRUPATH plasmid kit consists of 20 plasmids encoding 14 alpha, 2 beta, and 4 ...inhibition). AAV preps from DREADD plasmids allow you to easily express a DREADD receptor in a specific ...quick guide to chemogenetics Resources on Addgene.org Check out Addgene's chemogenetics guide Find chemogenetics...response? Let’s take a look at a few examples with plasmids deposited at Addgene.  PRESTO-TANGO: The ligands...resource called PRESTO-TANGO (parallel receptorome expression and screening via transcriptional output - TANGO...factor from the receptor, which can then activate expression of a luciferase reporter in the nucleus. The ...Bryan Roth’s lab also assembled a collection of plasmids to elucidate the role of 14 of the 16 non-visual...

  17. What's New in CRISPR - September 2019

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    Blog Post
    ...Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...an extensive RNA scaffold and a domain needed for DNA unwinding. They’ve deposited plasmids for active ...for E. coli and human expression. Read the paper in Nature Find the CasX plasmids Drug inducible transcriptional...vector sequence. Homology arms that flank the donor plasmid cargo are complementary to either side of the double-stranded... Acids Research Find the drug inducible Cas9 plasmids E. coli genome-wide inhibition library In the...targeting requires two sgRNAS: one that targets the knock-in site and a universal sgRNA that targets the donor...

  18. Lentiviral Vector Uses and Overview

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    Blog Post
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with... make stable transgene-expressing or knockout cell lines. Many CRISPR plasmids are designed for lentiviral... of Addgene’s most requested plasmids are used with lentiviral expression systems. If you’re new to the...lentivirus, check out our lentiviral plasmid and biosafety guides before beginning your experiment. Let...into the genome, permitting long-term transgene expression, but they could only infect actively dividing...non-dividing cells, but without sustained transgene expression. To design a viral vector system that could do...

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

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    Blog Post
    ...domain. These mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 is then tethered...et al., 2014). The sgRNA or tracrRNA sequences can also be modified to include RNA domains that recruit...Sanson et al., 2018) . This tracrRNA was combined with the sgRNA expression vector allowing the screen ...CRISPRa) to regulate gene expression. Some CRISPR libraries contain many sgRNAs per gene to increase confidence...Calabrese: human CRISPRa sgRNA library In addition to inhibiting gene expression, dCas9 can also be used... that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA aptamers, and the  MS2-P65-HSF1 helper ...screens can help reveal the function of lowly expressed genes or genes that are more drastically affected...

  20. Adenoviral Vector Production and Troubleshooting

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    Blog Post
    ...recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide webpage, and references...Resources on Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page  ...death of transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of ...capacity is limited to 8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune response... capacity (up to 36 kb!), long-term transgene expression and negligible toxicity. However, their production...down into 5 key steps: Construction of the rAdV plasmid (~1 week): The AdEasyTM system is the most popular...adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the transgene of interest...
Showing: 101 - 120 of 278 results