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We narrowed to 227 results for: cas9 expression plasmid

Showing: 141 - 160 of 227 results

  1. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...screens Differentially expressed genes  As the costs for global gene expression analyses like RNAseq are...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...sequencing to identify which gRNAs are differentially expressed in the separate groups and, as a consequence, ...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...activity and minimize off-target effects of CRISPR-Cas9." Nature biotechnology 34.2 (2016): 184-191. PubMed...

  2. Tips for arabidopsis transformation

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    Blog Post
    ...plant plasmid resources at Addgene  Choosing an expression system Choosing an appropriate expression vector....org Find plant plasmids Visit our plant resource page Deposit arabidopsis plasmids with Addgene ...this method include: Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a...options, but here are a couple of my favorites. Plasmids from the Tsuyoshi Nakagawa lab are binary gateway...prefer Golden Gate assembly, there are a series of plasmids from the Dinneny lab that additionally allow for...for visual selection of transformants based on expression of RFP in the seed (Emami, Yee and Dinneny, 2013...culture you are growing definitely contains your plasmid. Grow lots of Agrobacterium: The more agrobacterium...

  3. CRISPR Plasmids - Empty gRNA Vectors

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    Collection
    ...Whether the plasmid contains Cas9 and if so, which function of Cas9. We have plasmids with Cas9 that can ...by the Doudna Lab . Plasmid Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI... on the plasmid. gRNA plasmids that do not co-express a Cas protein require a separate plasmid that does...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...Mammalian Lentiviral expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent...Vectors CRISPR Plasmids: Empty gRNA Expression Vectors Browse CRISPR Plasmids By Function Genome Editing Cut...

  4. CRISPR Plasmids - C. elegans

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    Collection
    ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...none, need Cas9 plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription none, need Cas9 plasmid Boxem 47912...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...need Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid... elegans none, need Cas9 plasmid Ristow Do you have suggestions for other plasmids that should be added...specific locus. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA... gRNA expression plasmid based on factors such as selectable marker or cloning method. When using CRISPR...

  5. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    ...lactose released activates expression of GFP from the lac operon. GFP expression can then be used to determine...fantastic tools it’s now easy to monitor protein expression, localization, and protein-protein interactions... Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs...Addgene.org Find Fluorescent Protein Biosensors Browse Plasmids from the Tsien Lab Browse the Michael Davidson...

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

    Type
    Blog Post
    ...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation...recombination, RFP is expressed. Cre can mediate one of two deletions to allow YFP or M-CFP expression; these deletions...research? Check out Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0...1.0, 1.1, 2.1 plasmids Brainbow 3.0, 3.1, 3.2 plasmids References: Transgenic strategies for combinatorial...ways to spatially and temporally control gene expression, especially in in vivo models, and new Cre-lox... of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need a quick primer on how...fluorescent protein in the construct, OFP (orange), is expressed when Cre is absent; stochastic Cre excision results...

  7. CRISPR Plasmids - Drosophila

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    Collection
    ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...transcription Virmilion none, need Cas9 plasmid Bullock and Port 49330 pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin..., need Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid...CRISPR Plasmids: Drosophila Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime...homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A nickase fused to...specific locus. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA...

  8. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...genome and stable expression of the transgene   Transfer plasmids The transfer plasmid is the vector you...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes...crossover event between the vector plasmid and the packaging plasmid caused the Psi sequence to be placed... facilitates nuclear export) is expressed from a separate plasmid, and the promoter of the 5’LTR has been...(tTA, also expressed by your transfer vector) binds to the TRE and activates the expression of your gene...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...vectors fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral...

  9. Hot Plasmids - November 2023

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    Blog Post
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. .... Today, it's time for another edition of Hot Plasmids!  Here's what you'll find in this post:  New...edits, such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing efficiencies...., 2023 under CC-BY 4.0 License. Find PE6 plasmids here!  Doman, J.L., Pandey, S., et al. (2023)....therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific...pulse-chase experiments, and more. Find LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID...LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments open up new possibilities...

  10. Worm Expression Resources

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    Collection
    ... Plasmid Collections Worm Plasmids Worm Plasmids and Resources Plasmid Tools Worm Constructs...some of the plasmids and tools available at Addgene for use in worms. Browse Plasmids Plasmid Tools for ... Gene/Insert PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids that contain a worm...Addgene's worm CRISPR resources include plasmids expressing Cas9, empty gRNA backbones, and constructs ... genes by feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily engineer specific...backbones, fluorescent proteins, and Cre expression plasmids. Large collections or kits are described ...complete list of plasmids that can be used in worms, search Addgene's entire collection. ID Plasmid Description...

  11. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...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...how to generate the final recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide...2.*Pro-Tip*: once the correct recombinant pAdV plasmid is identified, re-transform into a bacterial strain...

  12. Overcoming the Challenges of Lentiviral Production

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    Blog Post
    ... multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually...encodes VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer...Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...and packaging plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection... a usable titer, or fine-tuning selection and expression in your target cell line, researchers often find...By dividing the viral elements across multiple plasmids, the risk of recombination and consequently, the...extra level of safety, they also require a fourth plasmid. The requirement for additional DNA can impact ...

  13. Lentiviral Prep Service

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    Collection
    ... 52962 lentiCas9-Blast Sp Cas9 Cut Blasticidin Expresses human codon-optimized S. pyogenes Cas9 protein...backbone. Zhang Cas9 and Accessories for Activating Gene Expression Catalytically-dead Cas9 (dCas9) can be fused...Root Cas9 Lentivirus Cas9 viruses can be used to interrogate the genome in a variety of ways. Cas9 proteins...CRISPR plasmids resource page . Cas9 for Editing Genomic Sequences To perform genome edits, Cas9 nuclease...Libraries Cas9 Pooled Barcoding Libraries Control Addgene's lentiviruses are made from plasmids that are... list of gRNA plasmids targeting specific genes. For more information on CRISPR plasmids and function,... of available viruses based on plasmids in our repository that express fluorescent proteins. ID Name Insert...

  14. Plant Plasmids and Resources

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    Collection
    ... Plasmid Collections Plant Plasmids Plant Plasmids and Resources...for a full selection of plasmids expressing different functional variants of Cas9 and empty gRNA backbones...creating plant expression vectors: A modular plasmid assembly kit for multigene expression, gene silencing...addition to individual plasmids, Addgene carries several useful toolkits and plasmid collections for use ...includes plasmids containing promoters that drive general and species-specific gene expression (see the... assembly of one or more gRNA expression cassettes - Qi-Jun Chen. Cas9 and gRNA backbone for use in rice...CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants - Niko Geldner. CRISPR-Cas9 vector...

  15. Plan Your Experiment

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    Collection
    .../or Cas9 expression or (2) a lack of efficient target cleavage in cells that do express both Cas9 and ...Figure 1). You will decide how to express Cas9, the delivery system for Cas9 and the guide RNA (gRNA), the...transfect, transduce, or may not express common promoters. Browse Cas9 plasmids for RNP delivery . As mentioned...non-homologous end joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity...insertions, by copying from a DNA donor template Cas9 or Cas9 fusions with proteins that promote HDR Requires...site. Consider alternative Cas9 enzymes with different PAM sequences or Cas9 enzymes with flexible PAM ...design. Addgene offers many plasmids containing validated gRNAs . These plasmids contain gRNAs that have ...

  16. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ... Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors ... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western blotting...and PCR, depending on the level of endogenous expression. We highly recommend the use of an electronic...ampules each of 4 clones with different levels of expression of our gene of interest. Advantages of the agarose...isolate individual cell clones with different expression characteristics from a population of transfected...Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org...

  17. CRISPR Plasmids - Bacteria

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    Collection
    ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert....2013.08.013. Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such as ...Ampicillin none, need Cas9 plasmid Qi Do you have suggestions for other plasmids that should be added ...population. ID Plasmid Promoter PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used ...Kanamycin none, need Cas9 plasmid Marraffini 42876 pCas9 BsaI E. coli, S. pneumoniae Chloramphenicol yes, cut...

  18. CRISPR Plasmids - Xenopus

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    Collection
    ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...transcription none, need Cas9 plasmid Chen Do you have suggestions for other plasmids that should be added...following CRISPR plasmids have been designed for use in Xenopus. Cut Fully functional Cas9 enzymes designed...surrounding the DSB is introduced along with the Cas9 and gRNA plasmids, the cell may instead repair the break ...genomic changes. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA... gRNA expression plasmid based on factors such as selectable marker or cloning method. When using CRISPR...nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with a ...

  19. CRISPR Guide

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    Collection
    ...Browse Plasmids: Purify Cas9 Alternatives for CRISPR Genome Engineering S. pyogenes Cas9 (SpCas9) is the...more about Cas9 multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...in which Cas9 is included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered...to either co-infect with a Cas9-producing lentivirus or use a Cas9-expressing stable cell line. After applying...strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...new tools. Figure 3: Overview of Cas9n Nickase and dead Cas9 The two Cas9 nuclease domains, RuvC and HNH...

  20. CRISPR References and Information

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    Collection
    ...sites for S. pyogenes Cas9, S. aureus Cas9, S. thermophilus Cas9, N. meningitidis Cas9, or Cas12a from your...vectors using ssDNA oligos p201G Cas9 ; p201B Cas9 ; p201H Cas9 ; p201N Cas9 ; pUC gRNA Shuttle PDF, 500 KB...CRISPR RNA array: Cas9 (pX260) or Cas9 D10A (pX334) ; tracrRNA: Cas9 (pX330) or Cas9 D10A (pX335) PDF,...KB Church gRNA design and cloning for Cas9 orthologs Cas9 plasmids PDF, 107 KB Chen and Wente Zebrafish... CRISPR plasmid summary for transcriptional activation dCas9 activators sgRNA dual expression: pAC2 , ...design and cloning All-in-one plasmid containing expression cassette for NmCas9 and BsmBI site for cloning...Addgene CRISPR Resources CRISPR Plasmids Addgene's collection of CRISPR plasmids. CRISPR Guide Addgene's guide...
Showing: 141 - 160 of 227 results