...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...

...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...3rd Varies 12,472 pairs Human SLC Activation Library 132561 Activation Human Superti-Furga 3rd 6 3,342 ...

...target gene Many different activators exist, including the multi-plasmid SAM system. Select Expression ...application. For example: To activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs...effective than dCas9 alone for mammalian cell lines. Activate (CRISPRa) Increase expression of an endogenous...without permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter...expression of CRISPR components Short window of CRISPR activity may decrease off-target effects Additional Resources...gRNAs based on predicted on-target and off-target activity A PAM sequence is absolutely necessary for Cas9...high-fidelity Cas enzyme. In addition to off-target activity , it is also important to consider factors that...

...Receptor Activating hM3D(Gq) DREADD rM3D(Gs) DREADD Inhibitory hM4D(Gi) DREADD KOR DREADD PSAM4 GlyR Promoter...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(s) PI Synapsin...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...

...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...

... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. Plasmid Gene/Insert Promoter...target RNA sequence, they adopt an enzymatically active state and can bind and cleave additional RNAs regardless... regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which ...cleaved. This non-specific cleavage is thought to activate programmed cell death or dormancy for phage-infected...

...icon . Receptor Activating hM3D (Gq) rM3D (Gs) PSAM-5HT3 Inhibitory hM4D (Gi) KORD PSAM-Gly Promoter Synapsin...(Gq) and rMD3(Gs) are activating, while hMD4(Gi) and KORD are inhibitory. PSAMs are engineered α7 nicotinic...types of PSAMs: 5HT3 coupled, which are activating, and Gly-coupled, which are inhibitory. Use the table below...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 ...ChR2HA:2a:PSAML141F,Y115F:GlyR PSAML141F,Y115F-GlyR CAG ChR2 2A Yes Sternson 32482 CAG::PSAMQ79G:GlyR-IRES-GFP...

...TALengineering Reagents Transcriptional Activators TALE Transcriptional Activator Expression Vectors You may also...TALE transcriptional activators. The Joung Lab has recently shown that TALE activators made on the framework...transcriptional activation domain at the carboxy-terminus of the protein. TALE activators expressed from...encoded in these plasmids robustly yields highly active artificial transcription factors ( Maeder et al...updated to enable identification of potential TALE activator binding sites in any given sequence of interest...plasmids required for FLASH assembly. ID Plasmid Activation Domain RVD of C-terminal 0.5 TALE repeat 43884...among the three methods, the final products are the same). The publicly available, web-based ZiFiT Targeter...

... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. Plasmid Gene/Insert Promoter...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...

... BbsI yes, activate, N-terminal S. pyogenes Zhang PX856 62888 Mammalian BbsI yes, activate, C-terminal...Bacteria BsaI yes, activate S. pyogenes Marraffini pWJ68 46571 Bacteria BsaI yes, activate S. pyogenes Marraffini...yes, activate S. pyogenes Luikart pRubiG-T2A-Cas9 75348 Mammalian/Retroviral from pXL yes, activate S. ... none S. pyogenes Boxem lentiSAMv2 75112 Mammalian/Lentiviral U6 yes, activate S. pyogenes Blast Zhang...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...-in-one vector, but their function (cut, nick, activate, interfere, etc.) is limited to that of the Cas...Cas9. We have plasmids with Cas9 that can cut, activate, interfere, or nick. Selection , such as Puromycin...

... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. Plasmid Gene/Insert Promoter...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...

... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. Plasmid Gene/Insert Promoter... none S. pyogenes Yang pBUN6A11 OsU3 BsaI yes, activate S. pyogenes Bar Chen pBUN6I11 OsU3 BsaI yes, interfere.... pyogenes Bar Chen pHSN6A01 AtU6-26 BsaI yes, activate S. pyogenes Hyg Chen pHSN6I01 AtU6-26 BsaI yes...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...

... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. Plasmid Gene/Insert Promoter...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...Selectable Marker PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of...

...) plasmids : Genome-wide screening of enhancer activity using a Firefly luciferase reporter. These plasmids... has increased stability and light output Sanjiv Sam Gambhir 104587 pHIV-iRFP720-E2A-Luc Firefly EF1α ...

...signal the presence of specific biomolecules, the activity of specific genes and cellular processes, or other...) High-performance calcium sensors for imaging activity in neuronal populations and microcompartments....GCaMP6f/m/s calcium sensors for imaging neural activity (Constitutive or Cre-dependent) Ultrasensitive...Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature. 2013 Jul 18;499(7458):295-300. Douglas ...expressing GCaMP6 and mRuby2 for imaging neural activity (GCaMP6f/m/s) (Constitutive or Cre-dependent) ...Cre-dependent) In vivo measurement of afferent activity with axon-specific calcium imaging. Nat Neurosci...

...fusion protein activates the expression of downstream genes. Multiple sgRNAs targeted to the same promoter ... either alone or fused to the transcriptional activator VP64. When directed to its target in gene promoters...promoter act synergistically to activate gene expression. The dCas9 protein serves as a negative control of the... the function of the VP64 activator and can also be used to inhibited gene expression via steric hindrance...These plasmids are described in: RNA-guided gene activation by CRISPR-Cas9-based transcription factors. Perez-Pinera...VP64 : Expresses inactivated S. pyogenes dCas9 (D10A, H840A) fused to VP64 transactivator domain in mammalian...below: ID Plasmid 47106 pcDNA-dCas9 : Expresses inactivated S. pyogenes dCas9 (D10A, H840A) in mammalian ...

... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. Plasmid Gene/Insert Promoter...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...Selectable Marker PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of...Marker PI Publication Visualize A catalytically inactive Cas9 (dCas9) fused to a fluorescent protein (FP...

... scientists have developed constructs to activate/inactivate genes when Cre is present. By expressing ...addition of an exogenous ligand (e.g. tamoxifen) to activate Cre. One advantage of this system is tight temporal.... Cre may be expressed globally under a broadly active promoter like CAG, or expressed only in a subset...Pu 87694 pCAG-ERT2-PhoCl-Cre-PhoCl-ERT2 light-activated Cre cTnT Mammalian Campbell 89573 pCAG-iCre iCre...expression. Depending on the construct, Cre may activate or repress gene expression. Common construct types...Description Expression System PI 11578 pSico Cre activates shRNA expression, removal of EGFP; See also similar... Lentiviral Jacks 11584 Lox-Stop-Lox TOPO Cre activates gene of interest by removing Stop sequences; Puro...

...Cas9 based approach to knockout, repress, and activate gene targets. To accomplish this they have taken...sequence repression dCas9-VPR fusion for gene activation CRISPR/Cas9 Plasmid Variants Schematic of the...Plasmids 175571 and 175572 express dCas9-VPR for activation. Usage Guide sequences can be designed using ...sequence: Up to two guides can be expressed from the same plasmid by utilizing one of the dual guide shuttle...

...particular protein can activate expression from that element. When this activation requires a transcription... much about its function and/or alter cellular activity in a way that’s interesting from an applied perspective...more subtle control over protein expression and activity thereby limiting the use of cellular resources... a green light responsive sensor protein that activates expression of lacZ in response to green light....interact in a bacterial two-hybrid assay, they activate expression of the reporter gene. Plasmid ID Promoter...fusion protein in pACλCI-β-flap results in the activation of lacZ expression from FW102 OL2-62 . Wolfe ...promoter region cloned into pH3U3-zif268 (omega) activates expression of ura3 and his3. In bacterial strain...