We narrowed to 102 results for: cas9 expression construct

...has a low quantum yield, and when transiently expressed in cells, generates much less signal than comparable...permit five-color fluorescent imaging! These eNL constructs use the brightest characterized luciferase, NanoLuc...photobleaching and phototoxicity are a concern, these constructs come in very handy. This approach is especially... endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent signal for imaging ...

...interest. To get the molecular components of CRISPR/Cas9 (Cas9 protein and the crRNA-tracrRNA complex) and the...regulatory control. But even with the advent of CRISPR/Cas9 technology, inserting large tags into a precise ...We’ve created and tested plasmids that use CRISPR/Cas9 to endogenously tag a wide variety of genes with...guidance on how to make your own gene-tagging constructs below. Read on to find out how these plasmids...process following the double strand break CRISPR/Cas9 makes at the target site. The whole segment—a GFP...the plasmid from being cut and destroyed by CRISPR/Cas9 during transfection. Making the double strand break...the fluorescent tag to the cells, we used CRISPR/Cas9 to make a precise cut in the genome near either ...

...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...

... while also adding additional filters such as expression, vector type, species, and more. Plasmids Browse...for Golden Gate assembly. MoClo Toolkit Plant Expression Sylvestre Marillonnet 95 plasmids for cloning...standardized parts to create eukaryotic multigene constructs. ONE-GO Biosensors Kit Fluorescent Proteins Mikel...Stiewe Use these donor vector libraries for CRISPR/Cas9-mediated saturation genome editing of human TP53...

...Yeast Expression, Insect Expression Chen Davidovich 41 plasmids to construct multigene expression vectors...assembly of CRISPR/Cas9 constructs for use in plants. The constructs are made with zCas9i, a high efficiency...multi-gene constructs for S. cerevisiae expression. Multiplex Yeast Toolkit (MYT) Yeast Expression, CRISPR...KTK) Bacterial Expression Tom Ellis DNA parts to create multigene constructs expressed in the cellulose-producing...4G-Cloning Plasmid Kit Bacterial Expression, Insect Expression, Mammalian Expression Stephan Gruber Platform for... Mammalian Expression Takashi Yamamoto Plasmids for construction of all-in-one CRISPR/Cas9 vectors expressing...Assembly Kit Other, Yeast Expression, Plant Expression, Bacterial expression Christopher Dupont 95 plasmids...

... 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...more Cas9 fusion tools later in the guide. Figure 3: Overview of Cas9n. Increasing fidelity of Cas9 Researchers...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...

...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...

...astrocyte-selective expression in AAV Dual promoter Separate expression pRGEB31 - Deliver sgRNA and Cas9 into plants... to express a gene; however, we have included a function section for if you want to stably express an ...genetic element, or want to express shRNA. Choose by: Species-Specific Expression Epitope Tag or Fusion Protein...Selectable Markers Species-Specific Expression If you want to drive expression of your favorite gene, you will...EF1a, CAG, Ubc Transient expression - Gradia Lab plasmids for mammalian expression, Ligation Independent ...Empty mammalian expression plasmid for cloning untagged proteins pcDNA3.1-mCherry - Expresses mCherry tag ... Inducible Expression page Bacteria Lac, T7, araBAD, trp mTagBFP2-pBAD - Protein expression vector with...

...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...

...(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...

... 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...

...tet-inducible Cas9-2A-EGFP expression. Based on LentiCRISPR v2 . Adam Karpf 167935 pLenti-tetON-KRAB-dCas9-DHFR-EF1a-TagRFP...Tetracycline (Tet) Inducible Expression Tetracycline (Tet) Inducible Expression Tetracycline Off Tetracycline...factors to further minimize background expression. Improved expression of the transactivator proteins in mammalian...Experimental Considerations Expression System If your gene of interest should only be expressed occasionally, use...Backbones Find backbones or constructs to use for cloning to control expression of your gene of interest....Top Transactivators (tTA or rtTA) Find a construct that expresses the transactivator for your tetracycline...PB-EF1a-TetOn3G PiggyBac vector expressing Tet-On 3G transactivator constitutively expressed by EF1a promoter Tet-On...

...dsDNA construct flanked by ITRs. Cells from the packaging HEK293 or HER911 cell lines expressing E1 are...pAdTrack series contains an IRES-GFP construct that enables co-expression of GFP with the transgene of interest...transgenes to express for a shorter period of time or when needing to package larger cargo. CRISPR-Cas9 Delivery...case of some commonly used Cas9 homologs, especially when having to express them alongside a guide RNA...capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral particle...L2, L3, L4, L5) regions. Early transcripts are expressed shortly after the virus enters the host cell, ... host cell manipulation. Late transcripts are expressed after DNA replication starts and primarily encode...

...the host genome can have on transgene expression and the expression of neighboring genes. rAAV episomes ...Addgene. CRISPR/Cas9 Delivery rAAV vectors are a popular choice for the delivery of CRISPR/Cas9 elements, making... sequence overlap. Co-expression induces homologous recombination and expression of the full-length transgene... genome editing. Packaging the Cas9 from Streptococcus pyogenes (SpCas9) and a gRNA together into an rAAV...this obstacle, has been to use smaller Cas9 orthologs, such as SaCas9 from Staphylococcus aureus . The Feng...splice together, reconstructing the full-length transgene that can then be expressed. This method allows...deliver genetic cargo into cells. Recombinant AAV constructs are designed by replacing the Rep and Cap regions...

... Plasmid Collections Bacterial Expression Systems Bacterial Expression Systems Protein Purification Protein...bacterial expression systems. Read below and browse our curated collection of bacterial expression plasmids...Addgene Blog E. Coli Strains for Protein Expression Protein Expression Epitope Tags Protein Tags Return to ...to top Controlled Expression It isn’t always advantageous to constitutively express your gene of interest...turn on expression only at a specific time. This collection contains plasmids whose expression levels ...Addgene Controlled Expression Resources Check out our Tetracycline (Tet) Inducible Expression Collection for...systems. ID Plasmid Promoter Inducer Expression Species PI 44249 pdCas9-bacteria pTetO Anhydrotetracycline...

...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...

... 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...more Cas9 fusion tools later in the guide. Figure 3: Overview of Cas9n. Increasing fidelity of Cas9 Researchers...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...