...SpCas9-NG - NG; increase in vitro activity SpG - NGN; increase nuclease activity SpRY - NRN/NYN, where R ...nuclease activity Cas9 enzymes will likely improve as more key residues for specificity and activity are discovered...select for gRNAs with high on-target activity and low off-target activity. Knockout libraries often target...engineered to increase specificity and nuclease activity available. Engineered Cas9s have also led to reduced...off-target effects Sniper-Cas9 - less off-target activity; compatible with truncated gRNAs to increase specificity...about Cas9 variants . Anti-CRISPR Prolonged Cas9 activity can increase the risk of off-target cutting or...(Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved in phages and other...

...viral delivery RNA delivery of Cas9 and gRNA In vitro transcription reactions generate mature Cas9 mRNA...ribonucleoprotein (RNP) complexes Purified Cas9 protein and in vitro transcribed gRNA are combined to form a Cas9-gRNA...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...cleavage of the desired target sequence or on-target activity . Two gRNA targeting sequences with 100% homology...based on their predicted on-target and off-target activity ( see gRNA design software ). Additionally, many...

...quality control, including AAV titration by ddPCR, in vitro and (when possible) in vivo testing, and full sequencing...chloride, 20 mM Tris at pH 8.0) Suitable for in vitro as well as in vivo delivery where the vector is ...by the physiologic body fluids, such as blood, vitreous, CSF, etc. Not suitable as a buffer for vectors...enhanced transduction of the retina following intravitreal injection in mice and marmosets as compared ...enhanced transduction of the retina following intravitreal injection in mice and macaque. This serotype... citations include: Scalabrino, et al. 2015. Intravitreal delivery of a novel AAV vector targets ON bipolar...

...FlipGFP-based activity reporter of SARS-CoV-2 main protease Mpro. (Unpublished) A FlipGFP-based activity reporter...that have been used for measuring viral entry and activity in neutralization assays and links to popular ... cleaves hACE2, potentially increasing viral infectivity.( 6 ) PCP4 - (Purkinje cell protein 4) is involved...Broad Institute has utilized the natural RNAse activity of the Cas13 protein to develop and optimize the... the method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2). The...

...experiments, including neuronal connectivity labeling. ID Name Expression System Activity Serotype PI 51509 AAV ...expression systems . ID Name Expression System Activity Serotype PI 99118 pAAV-CAG-tTA CAG-driven, constitutive...used for viral TRAP). ID Name Expression System Activity Serotype PI 98747 pAAV-FLEX-EGFPL10a EF1a-driven...used for cell ablation. ID Name Expression System Activity Serotype PI 124364 pAAV-FLEX-DTR-GFP CBA-driven...

...promoter. RCIscript-GoldyTALEN is designed for in vitro synthesis of TALEN mRNAs. Both 5’ and 3’ Xenopus...the strong CAG promoter or can be achieved by in vitro mRNA synthesis from the T7 promoter. Truncations...BioSciences (N153AA, C63AA) and showed robust cleavage activity in several later studies. pCAG-T7-TALEN(Sangamo... assay, which enables the evaluation of TALEN activities in cultured cells. Golden Gate TALEN 2.0 47388... included for generating TALEN mRNAs through in vitro transcription as an alternative. Based on transient...

...produce large amounts of this protein and study it in vitro ? These, in addition to many questions about bacteria... 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 wide range of highly controlled conditions in vitro , or, if you want to determine its structure using...EsaR derivatives found herein have different sensitivities to 3OC6HSL. The EsaR-responsive promoter can...

...49410 pCFD3-dU6:3gRNA dU6:3 BbsI Injection or in vitro transcription Virmilion none, need Cas9 plasmid ...3tandemgRNAs dU6:1 and dU6:3 BbsI Injection or in vitro transcription Virmilion none, need Cas9 plasmid ...49408 pCFD1-dU6:1gRNA dU6:1 BbsI Injection or in vitro transcription Virmilion none, need Cas9 plasmid ...49409 pCFD2-dU6:2gRNA dU6:2 BbsI Injection or in vitro transcription Virmilion none, need Cas9 plasmid ...

...177670 Inhibition Human Adamson 3rd ∼3 366 or 1,573 Activity-optimized genome-wide library Discontinued Knockout...Knockout Human Sabatini and Lander 3rd 10 178,896 Activity-optimized genome-wide library 1000000100 Knockout...Knockout Human Turner 3rd 10 8,260 Two plasmid human activity-optimized genome-wide library 1000000095 Knockout...Sabatini and Lander 3rd 10 187,536 Two plasmid mouse activity-optimized genome-wide library 1000000096 Knockout...

... the emission of photons. The robust signal, sensitivity, wide dynamic range, and resistance to photobleaching...in assays ranging from use as a reporter gene in vitro and in vivo to use in biosensors. The most common...) plasmids : Genome-wide screening of enhancer activity using a Firefly luciferase reporter. These plasmids...screening of transformants in cloning process. Lynne Postovit 119760 pSGDlucV3.0 Firefly/Renilla Insertion of...

...the antigen sequence, and cross-species reactivity. Reactivity is based on validation data from Addgene...Recombinant Antibody Description Source Species Isotype Reactivity Recommended Applications PI Return to Top Do ...

...Antibodies ID Recombinant Antibody Description Isotype Reactivity Recommended Applications Integrin Antibodies ...Antibodies ID Recombinant Antibody Description Isotype Reactivity Recommended Applications Browse Epitope Tag Antibodies.... ID Recombinant Antibody Description Isotype Reactivity Recommended Applications Browse Antibody Plasmids...

...using these opsins as a tool to control neuronal activity, scientists generally attach a fluorescent protein...protein interactions, dimerization, localization or activity of many different types of proteins. For a full...that use light to control a protein or cellular activity. For example, search "Cdc42", "transcription",...

...may reduce the gRNA activity. The variety in sequence decreases the gRNA activity and increases off-target...CRISPR systems with wiki entries describing enzyme activity, PAM requirements, experimental considerations...

... In vitro transcription none, need Cas9 plasmid Boxem 47912 SP6-sgRNA-scaffold SP6 AflII In vitro transcription...R07E5.16 U6 yes, cut Goldstein 42250 DR274 T7 BsaI In vitro transcription none, need Cas9 plasmid Joung 46170...

...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...

...promoters. These AAV can be used to compare the activity of different serotypes. Serotype AAV1 AAV2 AAV5...switch Brainbow constructs Spaghetti monster tags Activity Cre-dependent Flp-dependent Cre and Flp-dependent...Clear Filters ID Name Promoter Fluorophore/Tag Activity Serotype PI 37825 AAV-CAG-GFP CAG GFP Constitutive...

...Mammalian/in vitro transcription BsaI none S. pyogenes Mark pU6T7 71462 Mammalian/in vitro transcription...CRISPR systems with wiki entries describing enzyme activity, experimental considerations, and more, created..._LacZ 74293 Mammalian S. pyogenes Neo, mCherry Postovit pdCas9-DNMT3A-PuroR_v2 74407 Mammalian U6 yes,...

...(~25 kD), but tend to have lower affinity and longevity. Single-domain antibodies - Single-domain antibodies...monoclonal antibodies tend to have less cross-reactivity and give lower background signals than polyclonal...interest is knocked down or out, and antibody reactivity is compared between the wild-type and the knockdown...cell line with low or no expression; antibody reactivity is then compared between the knock-in cell line...

... Optogenetic tools can be used to control cell activity in living tissue with light. Optogenetics is typically...fluorescent protein Blue-wavelength fluorescent protein Activity Cre dependent Flp dependent Cre and Flp dependent...Clear Filters ID Name Promoter Opsin Variant Tag Activity Serotype PI 20071 pACAGW-ChR2-Venus-AAV CAG ChR2...