... 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...plasmids resource page . Cas9 for Editing Genomic Sequences To perform genome edits, Cas9 nuclease is used to...Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A ...Addgene's viral vector packaging service. Browse constructs currently available in lentiviral format. Viral...Browse In-Stock Lentivirus Pooled CRISPR Libraries Cas9 Pooled Barcoding Libraries Control Addgene's lentiviruses...

...resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of... Using CRISPR/Cas9 - Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) expression plasmids - Keith...Lab CRISPR/Cas9-based conditional mutagenesis in zebrafish - Wenbiao Chen Lab A CRISPR/Cas9 Vector System...fusion expression in organisms or cells in which an exogenous mRNA can be injected and expressed. EXPRESs...in zebrafish. Zebrafish expression atlas - A zebrafish-specific gene expression database that provides ...modified CRISPR-Cas9 system - Shuo Lin Lab Re-purposed Ac/Ds transgenic system enabling CRISPR/dCas9-interference...morpholinos, zinc finger nucleases, TALENS, and CRISPR/Cas9-based gene editing make the zebrafish a stalwart...

...have deposited the best performing sensors and constructs encoding corresponding portions of the Zika virus...virus research community. Find the Zika Sensor Constructs at Addgene As we report, we have embedded this...amplification, new toehold switch designs, CRISPR/Cas9-mediated genotyping, as well as our electronic reader...switches: de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939. PubMed PMID: 25417166...

... 2 (Th2) cells from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation...differentiation and activation, using expression of differentiation factors as the screen readout. This retroviral... voltage changes in worms and have made these constructs available at Addgene. These include Arch(D95N...Wang Lab created a tripartite system consisting of dCas9, a gRNA fused to an RNA binding motif, and a RNA-binding...protein-bEFP fusion. Thus, the bEFPs bind the UAS via the dCas9-gRNA complex which allows transcription from the...

...the Doudna Lab . Plasmid Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI pCRISPR...Mammalian Lentiviral expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent... plasmid contains Cas9 and if so, which function of Cas9. We have plasmids with Cas9 that can cut, activate...be used to express multiple gRNAs from a single plasmid with or without expression of Cas9. Highlighted...system for constructing all-in-one expression vectors containing multiple guide RNA expression cassettes.... Can express Cas9 with up to 3 gRNAs. Frew pSQT1313 Mammalian Single plasmid for the expression of multipe...cassettes are assembled into a Cas9 expressing vector. Yang A Robust CRISPR/Cas9 System for Convenient, High-Efficiency...

...Technique PI Purpose pCas9 42876 CRISPR Luciano Marraffini Bacterial expression of Cas9 nuclease gRNA. For...so multiple constructs can be made and tested at once. Return to Top Controlled Expression It isn’t always... constitutively express your gene of interest at high levels. Perhaps high expression leads to slow bacterial... on expression only at a specific time. This collection has a variety of plasmids whose expression levels...Anhydrotetracycline inducible expression of catalytically inactive Cas9 ( S. pyogenes ) which can be combined... bacterial expression systems to find out more! Genome Engineering Although overexpressing or knocking...see here . pwtCas9-bacteria 44250 CRISPR Stanley Qi Anhydrotetracycline inducible expression of wild-type...

...to create transgenic mice expressing fluorescent protein reporter constructs in the immune lineage. By...loxP sites such that their expression is turned on or off in cells expressing Cre. The result of a system...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... efficiently in two-photon microscopy and new constructs have facilitated the creation of mouse lines ...other technologies such as optogenetics and CRISPR/Cas9 will help immunologists create even better tools...

..., a base editor containing high fidelity Cas9 variant HF-Cas9 (Rees et al., 2017). HF-BE3 showed 37-fold...other methods. In a head-to-head comparison with Cas9, Cas9 modified 9/12 known off-targets with a 14% indel...individual Cas9. They also mutagenized the cytidine deaminase portion of the base editor to create SpCas9 base...et al., 2020 SpRY Cas9, available fused to ABEmax(7.10) or to BE4max   Evolved SpCas9 with NRN or NYN PAM... adenine base editors by expression optimization and ancestral reconstruction. Nature Biotechnology, 36... 1.1%. These numbers are a vast improvement over Cas9-mediated HDR for the loci tested, where average ...system is also subject to off-target editing based on Cas9 off-target activity.  Find CRISPR base editing plasmids...

... of expression. For example, CAG is a strong promoter that drives very high, ubiquitous expression, while...knowledge to be able to design a construct for the successful and specific expression of your gene of interest...such as SaCas9, CRISPR gRNAs, or your gene of interest. Promoter The promoter drives the expression of the... prevent ITR loss, it’s best to propagate AAV constructs in recombination-deficient bacterial strains,...type-specific, which can help you restrict the expression of your transgene to a specific species or location...to spatially and temporally control transgene expression. Lox sites are directional 34 bp sequences that...nuclear export of mRNA and helps increase the expression of your transgene. The WPRE is located at the...

... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...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...enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined...application. Cas9 Orthologs While SpCas9 is the most commonly used Cas enzyme, additional Cas9 orthologs ...Additionally, constructs with Cas9 fused to proteins that modulate HDR or NHEJ components, like miCas9 , Cas9...

...(tTA, also expressed by your transfer vector) binds to the TRE and activates the expression of your gene...the different vectors used, lentiviral vector constructs have proven very useful due to their ability ...vectors fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral...generation, the HIV Tat gene (previously used to drive expression from the LTRs) has been removed, Rev (which facilitates... facilitates nuclear export) is expressed from a separate plasmid, and the promoter of the 5’LTR has been...Stable integration into host genome and stable expression of the transgene   Transfer plasmids The transfer...a simple set-up where the gene of interest is expressed together with a marker (Fluorescent protein or...

...to fertility. Several zebrafish-specific gene expression databases that provide information about when...have been developed to help parse and visualize expression data in useful ways. Zebrafish mutants In the...zinc-finger nucleases, TALENs, and more recently CRISPR/Cas9. Another popular targeted gene deactivation method...sequencing-based lineage tracing Recently, gene expression in all cells of the developing zebrafish embryo..., relying on established reagents such as CRISPR/Cas9, Tol2, and the high-quality sequenced zebrafish ..., and their resistance (so far) to facile CRISPR/Cas9-mediated insertion of foreign DNA. Despite these...cells from 694 embryos over multiple stages and reconstructed the patterns of gene activity and cell divisions...

... while also adding additional filters such as expression, vector type, species, and more. Browse Browse...for Golden Gate assembly. MoClo Toolkit Plant Expression Sylvestre Marillonnet 95 plasmids for cloning...standardized parts to create eukaryotic multigene constructs. Pooled Libraries Pooled Library name Type PI...Stiewe Use these donor vector libraries for CRISPR/Cas9-mediated saturation genome editing of human TP53...

... vitro, and can be used for measuring protein expression or localization in cells, or even single-particle...background signal (especially for some older split-GFP constructs or certain cell environments). Also consider ...Split-horseradish peroxidase (Alice Ting Lab 2016)  Split-Cas9 for inducible genome editing (Feng Zhang Lab 2015...

...plasmids expressing HTT, C9ORF72, LRRK2, Synuclein, and more. New and Noteworthy: Use tau constructs to study...Neurodegeneration Plasmid Collection A collection of plasmids expressing wild type or mutant ORFs, gene fragments, and...Target neural oxytocin receptors using an AAV-CRISPR/Cas9 strategy for gene editing across divergent rodent...species. Boender et al. Sci Adv. 2023 Jun 2. Use Cas9 in astrocytes. Endo et al Science. 2022 Nov 4. Base... dopamine research. CBA-driven, Cre-dependent expression of diphtheria toxin receptor (DTR)–GFP fusion...2023 Dec 26. Induce CNS cells with concomitant expression of aggregation-prone proteins with the iPSC toolbox...Dec 2. Use PiggyBac plasmids with tet-inducible expression of transcription factors for sensory, motor, ...

...can then be expressed. This method allows for longer transgene expression, but expression is much less...integration can affect the expression of transgenes, as well as the expression of neighboring genes. The... sequence overlap. Co-expression induces homologous recombination and expression of the full-length transgene...cycle; VP1, VP2, VP3 - capsid proteins). When constructing an AAV transfer plasmid, the transgene is placed...plasmids available from Addgene. The popular CRISPR/Cas9 genome editing system has been modified for use ...Episomal stability enables long-term transgene expression in non-dividing cells and is a key advantage ...synthesize the complementary strand, transgene expression may be delayed. To overcome this rate-limiting...

... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...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...enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined...application. Cas9 Orthologs While SpCas9 is the most commonly used Cas enzyme, additional Cas9 orthologs ...Additionally, constructs with Cas9 fused to proteins that modulate HDR or NHEJ components, like miCas9 , Cas9...

...different versions of dCas9. Following integration, Mobile-CRISPRi is stably expressed with no selection ... system. This system has two parts: a plasmid expressing the Tn7 transposition genes (tnsA, tnsB, tnsC...efficiency, as measured by knockdown of a constitutely expressed RFP reporter, was variable across bacterial strains... a proof of principle, Peters and colleagues constructed a 40-member library targeting 20 genes of Enterobacter...induction of CRISPRi knockdown. As expected, bacteria expressing guides that target essential genes were less ...A Tn7-based broad-range bacterial cloning and expression system." Nature methods 2.6 (2005): 443. PubMed...antibiotic resistance selection markers, gRNAs, and dCas9 proteins. Second, it can be introduced into a broader...

...al., 2024). In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study...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 ...Fourier-shell correlation curve (D), and the reconstructed 3D map with 3.1 Å global resolution (E). Figure...