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We narrowed to 356 results for: SAM activators

Showing: 1 - 20 of 356 results

  1. Zhang Lab CRISPR Page

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    Collection
    ...Synergistic Activation Mediator (SAM) is an engineered protein complex for the transcriptional activation of endogenous...HSF1 activation helper protein Full references are below. Pooled genome-wide human SAM library SAM can ...please visit the Addgene SAM library page . Targeted gene activation using SAM There are two sets of 3 ...references are below. Return to top Cas9 SAM transcriptional activation plasmids and screening library CRISPR... available for mammalian endogenous gene activation using SAM: Addgene plasmids #61422 - #61424 were designed...Expresses dCAS9-VP64 activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with... Return to GeCKO Library SAM Library Genome-scale transcriptional activation by an engineered CRISPR-Cas9...

  2. CRISPR Pooled gRNA Libraries

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    Collection
    ...70,290 SAM v1 - 3 plasmid system 1000000075 (Puromycin) Activation Mouse Zhang 3rd 3 69,716 SAM v2 - 2... 2,852 Broad GPP activation Calabrese p65-HSF 92379 (Set A) 92380 (Set B) Activation Human Doench and ...(Set B) Broad GPP activation Caprano p65-HSF 92383 (Set A) 92384 (Set B) Activation Mouse Doench and Root...Discontinued Activation Human Weissman 3rd 10 198,810 CRISPRa-v2 83978 1000000091 Activation Human Weissman...15,416 Human CRISPR lncRNA Activation Pooled Library 1000000106 Activation Human Zhang 3rd 10 96,458 Human... 10 7,501 Human Membrane Protein Activation Library 113345 Activation Human Wright 3rd 7-14 58,570 Human...Sanjana 3rd 6–8 75,184 Human SLC Activation Library 132561 Activation Human Superti-Furga 3rd 6 3,342 ...

  3. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

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    Blog Post
    ...Synergistic Activation Mediator (SAM) Description SAM uses specially engineered sgRNAs to increase transcription...gene activation compared to other CRISPR activators, making it a popular method for gene activation experiments...gene regulation (activating multiple genes at once), however, SAM exhibits activation levels comparable...than first generation activators while showing lower activation levels than SAM. One drawback of this ...higher activation levels than the initial dCas9-VP64 activator, but lower levels compared to the SAM system...common choice for CRISPR activation. Newer methods for CRISPR activation While SAM, SunTag, and VPR have ...components of SAM and SunTag activators, fusing the epitope tag of SunTag with the P65-HSF activation domains...

  4. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    Blog Post
    ... they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM is an engineered protein complex for... the activation helper protein. SAM can be combined with a human genome-wide library to activate all 23,430...gene activation in mind. “We had not managed to create a system to allow us to reliably activate essentially...information about SAM, including a tool for getting optimized guide sequences for activating any human gene... leap from mutating and deactivating a gene or genes to selectively activating them with the CRISPR system...her then was this: Can you engineer CRISPR-Cas9 activators that work well enough on any gene that they could...genome-wide screening,” Konermann said. “If you want to activate all 20,000 genes, it is not possible to test each...

  5. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    Blog Post
    ...synergistic activation mediator (SAM) induces specific, persistent and robust reactivation of the HIV-... CRISPR-HIV therapeutics - one using dCas9-SAM to activate HIV-1 transcription and destroy infected cells...turned to the CRISPR/Cas9 Synergistic Activation Mediators (SAM) system available from Addgene. This system...fusion commonly used to activate transcription, but with two additional activation domains (MS2 and p65)...found that SAM targeting of the enhancer region near the NF-KB binding sites increased activation of HIV ...treatment with CRISPR/Cas9 SAM increased the percentage of cells with activated HIV-1, measured via HIV-...realistic with the advent of CRISPR. Using CRISPR/Cas9 SAM to "shock and kill" To “shock and kill” the HIV-...

  6. CRISPR Activation: A Practical Guide

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    Blog Post
    ... Cas9 deficient in DNA cutting activity to a transcriptional activation domain (Fig 1). Two ways to ... bound of activation that current Cas9 systems cannot help you pass. In addition, activation experiments...Cas9-based activation. CRISPRa for genome-wide studies One of the best uses for Cas9 activators is in genetic... it much cheaper to use Cas9 activators. This larger scale of activation allows for exploratory-based ...readily available. By activating from the native context of the gene, Cas9 activators efficiently solve these...problems.   Which Cas9 activator should I use? There are a wide variety of activators you can use for your...different types of Cas9 activators, check out our blog post covering VP64, Suntag, SAM, and VPR. Worry less...

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

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    Blog Post
    ...cells. The SAM system (CRISPR/Cas9 Synergistic Activation Mediator) uses an improved activator design that...dCas9 can also be used for transcriptional activation. Activation of specific genes allows scientists to ... nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or an activation domain (CRISPRa...Rational design of highly active sgRNAs for CRISPR-Cas9–mediated gene inactivation." Nature biotechnology...libraries can be used to knock out, inhibit or activate target genes by combining specific sgRNAs with...knock out), CRISPRi (interference), and CRISRPa (activation) libraries that do just that (Doench et al., ...identified in Doench et al., 2014 to predict sgRNA activity. In this study scientists examined approximately...

  8. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...website about star activity . If you are digesting a large number of plasmids with the same enzyme(s) (for... However, keep in mind that restriction enzyme activity is determined under ideal conditions with very...their recognition sites. This is due to "Star Activity" and can happen for a variety of reasons, including...double digest (digesting with two enzymes at the same time), you will need to determine the best buffer...will not be gel purifying it, you may need to inactivate the enzyme(s) following the digestion reaction...

  9. Ligation Independent Cloning

    Type
    Protocol
    ...exonuclease function of the enzyme and shift its activity back to polymerase. In our case, dGTP will be ...protocol), causing the enzyme to perform exonuclease activity until the first "G" in the sequence. Because the...proceeding, as they will interfere with the exonuclease activity of T4 Pol in the following step. This may be accomplished...have different leader sequences, but operate on the same principle (homologous to the first G on 3'-5' strand... incubate at room temperature for 30 minutes. Inactivate T4 Pol by heating to 75° for 20 minutes. Volume...

  10. Plasmids 101: The protein expression toolbox

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    Blog Post
    ...CRISPR for that, too! CRISPR activators (CRISPRa) are transcriptional activators fused to a dead Cas9. The...Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the time...increase expression. There are several flavors of activators which can be used with this approach - read more...them on our CRISPRa blog! Fig. 1 CRISPR activator and repressor mechanisms Want to transiently...

  11. Plan Your Experiment

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    ...fusion with transcriptional activator (such as dCas9-VP64) Many different activator systems exist, including...including the multi-plasmid SAM system Activate Design and Clone Your gRNA Achieving your desired edit comes...Knockouts Activation/interference Needs good primer design Western blot Knockouts Activation/interference...the gRNA activity. Be sure to sequence the genomic region you intend to target in your samples to ensure...proteins) Many base editors fused to Cas9 exert their activity within a very narrow window proximal to the PAM...dCas9 alone for mammalian cell lines Interfere Activate (CRISPRa) Increase gene expression without permanently...essential protein domain could abolish protein activity and essentially function as a “knockout,” but ...

  12. Luciferase Plasmid Collection

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    ... has increased stability and light output Sanjiv Sam Gambhir 104587 pHIV-iRFP720-E2A-Luc Firefly EF1α ... upon interaction, the NanoBiT subunits form an active enzyme, generating luminescence. Find plasmids ...and Tools Optimized STARR-seq (Self-Transcribing Active Regulatory Region) : Genome-wide screening of enhancers...G-CASE : BRET-based sensors for reporting on GPCR activity. AIMTOR : BRET biosensor for live imaging of mTOR...

  13. What's New in CRISPR - November 2021

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    Blog Post
    ...RNAs Yong-Sam Kim’s lab also created a miniature yet active Cas system starting with the same Un1Cas12f1...version (named CasMINI) had the highest activity for gene activation  and base editing, whereas a different...editing enzyme with a fully active Cas9 enzyme. PE-Cas9 comprises an active Cas9 fused to the reverse transcriptase...especially when fused to effector proteins for gene activation or gene editing. The more recently identified...would be ideal for such AAV packaging, but their activity in mammalian systems has been in question. Three...CasMINI system was shown to be effective for CRISPR activation, base editing, and genome editing in mammalian...version) works best for genome editing  with an active nuclease.  Read the paper in Molecular Cell Find...

  14. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    Blog Post
    ...website. Activating target genes using dCas9-based activators The simplest of the dCas9 activators consists...single transcriptional activation domain - VP64.  Later generations of activators include the SunTag system...tagged dCas9 and antibody-activator fusion proteins. The Synergistic Activation Mediator complex, generally...additional RNA-binding transcriptional activator. Plasmids for activating target genes in a variety of species...promoter region. Cas9 activators include Cas9 fused to VP64, VP64 along with other SAM factors, and the Suntag...facilitates recruitment of multiple activation domains to the same genetic locus via co-expression of ...researchers to knock out, knock in, base edit, activate, repress, and even image your favorite gene. With...

  15. Chemogenetics AAV Preps

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    ...hM3D(Gq) - Activation mCherry fusion Flp-dependent 8, rg* Gether 119741 AAV SYN flex PSAM4 GlyR IRES EGFP...chemogenetics AAV to chemically induce neuronal activity in specific cell types. See our Chemogenetics ...mCherry HA Non-fusion tags mCitrine EGFP dTomato Activity Cre-dependent Flp-dependent Cre and Flp-dependent...Retrograde AAV PHPeB Clear Filters ID Name Receptor and Activity Reporter/Tag Cre requirement Serotype PI 44361...44361 pAAV-hSyn-DIO-hM3D(Gq)-mCherry hM3D(Gq) - Activation mCherry fusion Cre-dependent 1, 2, 5, 8, 9, rg...pAAV-hSyn-DIO-HA-hM3D(Gq)-IRES-mCitrine hM3D(Gq) - Activation HA fusion and IRES mCitrine Cre-dependent 8 Roth...50458 pAAV-hSyn-DIO-rM3D(Gs)-mCherry rM3D(Gs) - Activation mCherry fusion Cre-dependent 8 Roth 50474 pAAV-hSyn-hM3D...

  16. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    Blog Post
    ...other hand, recruits transcriptional activators such as VP64, SAM, or SunTag to specific ncRNA loci to...DNA, CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) are powerful tools. CRISPRi uses a ...insertions or deletions in the DNA sequence. This same approach is effective to target short ncRNAs, such...useful for targeting thousands of long ncRNAs at the same time (Liu et al. 2018). However, removing large ...

  17. Chemogenetics Plasmids

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    ...light that activates the opsin to either excite or inhibit the target neuron. LMO can be activated either ...Exclusively Activated by Designer Drugs (DREADDs) and Pharmacologically Selective Actuator Modules (PSAMs). DREADDs...(Gq) and rM3D(Gs) are activating, while hM4D(Gi) and KORD are inhibitory. PSAMs are engineered α7 nicotinic...types of PSAMs: 5HT3 coupled, which are activating, and GlyR-coupled, which are inhibitory. Another recently...encode genetically engineered receptors that are activated by specific small molecules, allowing a scientist...expressed, these receptors can be used to artificially activate or inhibit specific biological processes. Addgene... activating or inhibitory depends on the ion pore domain it is coupled to. Addgene has two main types ...

  18. CRISPR Guide

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    ...helper activators; highest levels of single-gene activation (Figure 9D) In bacteria, activating gene expression...nuclease activity Cas9 enzymes will likely improve as more key residues for specificity and activity are discovered...-NG - NG; increase in vitro activity SpG - NGN; increase nuclease activity SpRY - NRN/NYN, where R = A...select for gRNAs with high on-target activity and low off-target activity. Knockout libraries often target...become a flexible tool to activate, repress, visualize, and isolate genes. Activation or Repression of Target...transcriptional activator (e.g. VP64) or repressor (e.g. KRAB; Figure 9A). Other examples of activation systems...instead of a two-component system (Figure 9C) SAM activators - co-expression of dCas9-VP64 with a modified...

  19. Lighting Up Cell Signaling with Photoswitchable Kinases

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    Blog Post
    ...and pa= photoactivation) domains on either side of the active site of a constitutively active MEK1. This...localization serves as an indicator of MEK1 activity. When MEK1 is active, it phosphorylates the ERK KTR and red...and gut cells of C. elegans causes the same phenotype. Activation of the Raf-MEK-ERK signaling pathway ...Protein Activity with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent...signaling networks can also be non-specifically activated and, like the game of telephone, often the signal...intermediate messengers. Optical control of kinase activity can provide greater spatiotemporal resolution ...system, light is used to turn “on” and “off” MEK1 activity by attaching two dimerizing photoswitchable Dronpa...

  20. Viral Vectors 101: Chemogenetics

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    Blog Post
    ...with less endogenous activation and reduced off target activation by ligands. PSAMs themselves are modified...engineered receptors that are only activated by otherwise inert drugs. Once activated, chemogenetic receptors hijack...First Structure–Activity Relationship Studies for Designer Receptors Exclusively Activated by Designer Drugs...DREADDs and PSAMs. Using DREADDs DREADDs DREADDS, or designer receptors exclusively activated by designer...provides a noninvasive method for manipulating cell activity, especially neuronal firing. What is chemogenetics...chemogenetics Because chemogenetic receptors are activated through administered drugs that must diffuse throughout... throughout the body, the onset of receptor activation is delayed and takes on the order of minutes. Studies...
Showing: 1 - 20 of 356 results