...fluorescence indicator of K+ for optical imaging (GINKO) Genetically encoded fluorescent indicators for imaging...comparison of a genetically encoded sensor and small molecule indicator: implications for quantification of cytosolic...Javier García-Sancho Calcium Genetically encoded calcium indicators for optical imaging (GECOs), including...and ratiometric indicators An Expanded Palette of Genetically Encoded Ca2+ Indicators. Science. 2011 Sep...Teal genetically encoded calcium sensor T-GECI Blue-shifted genetically encoded Ca(2+) indicator with ...fluorescent cAMP indicator (Flamindo2) Genetically-encoded yellow fluorescent cAMP indicator with an expanded...protein-based cAMP indicator (Pink Flamindo) Red fluorescent protein-based cAMP indicator applicable to optogenetics...

...Handling Visualization Signal Amplification Application Overview Quantification Methods Capture Methods Visualization...signaling molecules and are often used in clinical applications. Diabodies - Diabodies contain two Fab fragments...this can vary significantly based on the antibody and conjugate chosen. Signal Amplification The amount ...single assay. Overview of Antibody Applications Antibody-based applications can be generally classed into ...these applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...processing steps, such as sonication. Denature proteins, using heat and/or chemicals, and run on an SDS-PAGE...concentrates in the sample. Controls for quantification methods Quantification methods should be run with: a positive...

...using AAV for research is that it is replication-limited and typically not known to cause disease in humans...Cap controlled by tTA-TRE amplification system Gradinaru 103006 pUCmini-iCAP-PHP.S PHP.S AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 127847 pUCmini-iCAP-PHP.V1 PHP.V1 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 175004 pUCmini-iCAP-AAV.CAP-B10 B10 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 175005 pUCmini-iCAP-AAV.CAP-B22 B22 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 185136 pUCmini-iCAP-AAV.MaCPNS1 MaCPNS1 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 185137 pUCmini-iCAP-AAV.MaCPNS2 MaCPNS2 AAV packaging...

...immunoprecipitation (enChIP) using CRISPR. Biochemical and Biophysical Research Communications . 439 (1), 132–136. PMID:... organisms, but modifications to various Cas enzymes have extended CRISPR applications to increasingly...takes place during the S and G2 phases. Chemically or genetically inhibiting genes involved in NHEJ or upregulating...components, like miCas9 , Cas9-DN1S , or others, can increase HDR efficiency. In most applications, a large portion...genes. Each library typically contains ∼3–6 gRNAs per gene to ensure modification of every target gene...following the amplification protocol specified by the depositing lab. After amplification, you’ll need ...heritability. Not all modifications are stable across cell divisions, and some types of modifications are more frequently...

...Double floxed soma and AIS-localized genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production...-DIO-JEDI-2P-WPRE Double floxed genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production...-Kv-WPRE Soma and AIS-localized genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production...179464-AAV9 pAAV-hSyn-JEDI-2P-WPRE Genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production... the human brain through the development and application of innovative tools enabling large-scale real-time...or when BRAIN Initiative grants are noted in publications associated with Addgene materials. Plasmids ... 113049-AAV9 pAAV-hSyn-GRAB_DA1m express the genetically-encoded fluorescent dopamine(DA) sensor GRAB_...

...Insert Promoter Selectable Marker PI Publication Activate Catalytically dead dCas9 fused to a transcriptional...Insert Promoter Selectable Marker PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional...Selectable Marker PI Publication Epigenetics To make targeted epigenetic modifications, researchers have ... Gene/Insert Selectable Marker PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used...Insert Promoter Selectable Marker PI Publication Visualize A catalytically inactive Cas9 (dCas9) fused to a...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Browse CRISPR Plasmids By Function Genome Editing...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...

...therapeutic applications. Despite the ethical controversies surrounding non-research applications, it’s clear...just as useful (if not more so) for research applications, eclipsing past genome engineering technologies...to destroy the phages upon re-exposure. More specifically, specialized Cas proteins snip foreign DNA into...number/types of Cas proteins. Makarova et al. ’s classification defines 5 types and 16 subtypes based on shared...for CRISPR genomic editing. Genome Engineering Applications In 2012, Jinek et al. first demonstrated that... PubMed lists more than 6,300 CRISPR-related publications, many of which detail work to improve the tool...various species, as well as the development of new applications. The first CRISPR papers described two main ...

...resources for this meeting's publication, we are asking participants to share practical information in the form...permission to share (e.g., publication's permission). Connie Cepko, Harvard Medical School Ali Cetin and Shenqin...contribute to this publication. Citation information for openly shared communication: Adeno-Associated ...goal of this year’s workshop is to brainstorm practical strategies for using combinations of AAV-based...result of the meeting will be a publicly shared practical guide for all scientists in the field and every...Organizing Committee: Dr. Constance Cepko, Harvard Medical School Dr. Melina Fan, Dr. Susanna Bachle, Dr. ...Addgene Confirmed Speakers Constance Cepko Harvard Medical School Ali Cetin Allen Institute for Brain Science...

...Catalytically inactive CRISPR plasmids for purification of target loci....conventional methods for genomic purification, CRISPR-based purification methods are more straightforward and...Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast ID...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...chromatin immunoprecipitation (ChIP) to allow purification of any genomic sequence specified by a particular...DNA-binding molecule-mediated ChIP) system, catalytically inactive dCas9 is used to purify genomic DNA... can be fused to dCas9 or gRNA for efficient purification. Various epitope tags including 3xFLAG-tag, ...

...ID Plasmid Gene/Insert Promoter PI Publication Activate Catalytically dead dCas9 fused to a transcriptional... Plasmid Gene/Insert Promoter PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional...population. ID Plasmid Promoter PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used...than NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Browse CRISPR Plasmids By Function Genome Editing...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...gRNA Design Tools CRISPR Blog Posts Base Edit Catalytically dead dCas9 fused to a cytidine deaminase protein...opposite strand) mutations. ID Plasmid Promoter PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted...

...Manipulation Select Expression System Design gRNA Publications Get Started CRISPR is a powerful system that...may not be a perfect plasmid for your specific application, and in such cases, it may be necessary to customize...reagent to suit your needs. Genetic Manipulation Application Cas9 gRNA Additional Considerations Knockout ...method Expression System Components of System Application Mammalian expression vector Cas enzyme promoter...EF1alpha, CBh) or inducible (Tet-ON); U6 promoter is typically used for gRNA May contain reporter gene (e.g. ...gene you target will depend on your specific application. For example: To activate or repress a target...needs to match the target locus, but it is also critical to ensure that the gRNA target sequence does NOT...

...differentiate into multiple cell types, but are typically limited to related cell lineages. Pluripotent ...organism. iPSC technology removes the possible ethical concerns related to embryonic stem cell use and...generation of iPSCs is relatively simple in concept: ectopically express a cocktail of stem cell reprogramming...Blood Cells. Stem Cells. 2012 Nov 29. Yamanaka Replicating EBNA1 episome Human Non-integrating EBNA1-mediated... Nat Methods. 2011 May;8(5):409-12. Yamanaka Replicating EBNA1 episome Human Non-integrating polycistronic... S A. 2014 Jul 22;111(29):10678-83. Capecchi Replicating EBNA1 episome Human Fluorescent-tagged EBNA1-...Stem Cell Res Ther. 2017 Jun 5;8(1):132. Zovein Replicating EBNA1 episome Human Non-integrating EBNA1-mediated...

...of a catalytically inactive form of Cas9 (dCas9) plus guide RNA (gRNA) for biochemical purification (Fig... doi: 10.1093/dnares/dsx023. PubMed . Identification of physical interactions between genomic regions ...protein such as LexA are subjected to affinity purification (Fig. 1). In enChIP, specific genomic regions...analysis reveals genomic regulatory regions that physically interact with the Pax5 promoter in a chicken ...10.1111/gtc.12341. Epub 2016 Feb 5. PubMed . Identification of non-coding RNAs associated with telomeres...journal.pone.0123387. eCollection 2015. PubMed . A critical role of the Thy28-MYH9 axis in B cell-specific...26. doi: 10.1186/s12867-014-0026-0. PubMed . Identification of proteins associated with an IFNgamma-responsive...

... opens in a new window) ClinicalTrials.gov - a list of Rett syndrome clinical trials (Link opens in a ...window) PMID: 16615965 Leonard et al. 2017. Clinical and biological progress over 50 years in Rett syndrome...months of age. A defining feature, with onset typically between 18 to 30 months, is regression of previously...of Rett syndrome is currently based on specific clinical diagnostic criteria, defined as the presence of...MECP2 gene. Causative mutations for Rett syndrome typically arise spontaneously, although in rare cases Rett...determines a minimally functional MECP2 that rescues neurological defects of Rett syndrome. (Link opens in a new...Line Mutation (DNA) Background Strain Purpose Publication PI Mecp2 NLucTom Knock-in of NLuc-tdTomato at...

... the original publication below. You can also download the full-text of the publication here . Article...given the efficiency of biallelic modification, ease of rapid identification by PCR, predictability of loss-of-function...therapy and research. Gehl J. Acta Physiologica Scandinavica . 2003. PubMed . Purifying plasmid DNA... of Visualized Experiments (JoVE). The video publication by Stuart Orkin's and Daniel Bauer's labs details...addition, we describe techniques for deletion, identification, and characterization. This strategy relies...to help identify guide sequences that minimize identical genomic matches or near-matches to reduce risk...band” is often too large to efficiently amplify. Typically use primers at least 100 bp from the predicted...

... includes anti-glypicans that accurately detect glypican presence and explore glypican function, providing...about IPI (Link opens in a new window) Glypican Antibodies Glypicans are a family of highly conserved proteoglycans...Institute for Protein Innovation Antibody Collection Glypican Antibodies Integrin Antibodies Epitope Tag Antibodies...families of cell surface and secreted proteins (glypicans and integrins) and common epitope tags, as well...scientists worldwide. Figure courtesy of IPI Browse Glypican Antibodies ID Recombinant Antibody Description...Description Isotype Reactivity Recommended Applications Integrin Antibodies Integrins are a family of mechanotransduction...Antibody Description Isotype Reactivity Recommended Applications Browse Epitope Tag Antibodies IPI has developed...

...Insert Promoter Selectable Marker PI Publication Base Edit Catalytically dead dCas9 fused to a cytidine deaminase...Insert Promoter Selectable Marker PI Publication Activate Catalytically dead dCas9 fused to a transcriptional...Insert Promoter Selectable Marker PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional...Insert Promoter Selectable Marker PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression...

...Genome Engineering Visualization and Tagging Purification Controlled Expression Reporter Plasmids Reporter...Reporter Plasmids Tagging and Visualization Purification Controlled Expression Reporter Plasmids You may...Bacteria? E. coli are commonly used to store and replicate plasmids of all types, but beyond that, researchers...sites within this plasmid. Return to Top Protein Purification Sometimes looking at a protein inside cells ...and, epitope tags can be used to simplify the purification process. Using plasmids in this collection, ...those tags from the purified protein. Protein purification can be challenging, but once you get it right... 5% sucrose. Other empty vectors for protein purification from the Structural Genomics Consortium and ...

...Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter PI Publication Drosophila ID Plasmid...Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication C. elegans...Marker PI Publication Yeast ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Zebrafish...Marker PI Publication Xenopus ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Parasites...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Do you have suggestions for other plasmids that...

...make targeted epigenetic modifications, researchers have fused catalytically dead dCas9 to epigenetic ...Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Do you have...CRISPR plasmids for targeted epigenetic modification, including histone acetylation/demethylation, and...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...enhancer for your gene of interest. Available modifications include: histone acetylation by p300 histone...or MQ1 cytosine demethylation by Tet1 These modifications persist over time and are potentially heritable...tables below for CRISPR plasmids for epigenetic modification in mammalian or plant systems. Mammalian ID ...