...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 ...

...Gene/Insert 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...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 Prime Edit Cas9 H840A nickase fused to a reverse...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression...

... any application Balanced Salt Solution (BSS) An ophthalmic solution suitable for any application TMN200...When using the AAV2(Y444F) serotype in future publications, please acknowledge Arun Srivastava and cite...When using the AAV2(trpYF) serotype in future publications, please acknowledge Arun Srivastava and cite... using the AAV2(4pMut)dHS serotype in future publications, please acknowledge Shannon Boye and cite Boye...using the AAV6(dbY-F+T-V) serotype in future publications, please acknowledge Arun Srivastava and cite...Strategies for Overcoming Donor-Variation and Implications in Genome Editing. Sci Rep . Oct 19;6:35495....

...it's usually more efficient than plasmid donor. Typical design is to have 50-80bp of homology arms on each... changes (>100bp insertions or deletions), we typically use plasmid donor, with two homology arms on each...transfection in cells like HEK, we use around 400ng of a typical size (~ 5kb) donor plasmid with Cas9/sgRNAs vector...plasmids. For ssDNA oligo design, the Zhang lab typically uses around 100-150bp total homology. The mutation..., giving 50-75bp homology arms. The Zhang lab typically use PAGE purified long oligos. For large changes...100bp insertions or deletions), the Zhang lab typically uses a plasmid donor, with two homology arms on...like HEK, the Zhang lab uses around 400ng of a typical size (~5kb) donor plasmid with Cas9/sgRNAs vector...

...the most abundant forms of posttranslational modifications in cells and research into its many roles in...Yale have made improvements to this system by genomically recoding E.coli (Lajoie et al., Science 2013)...of the popular SepOTSλ ( 68292 ). An important technical note: pSerOTS-C1* ( 188537 ) is a ROP minus plasmid...Barber et al, Biochemistry 2018). Iterative Synthetically Phosphorylated Isomers (iSPI). This new resource...WW2 domain) References & Protocols Iterative Synthetically Phosphorylated Isomers (iSPI) described in: ...an orthogonal translation system with enhanced biological tolerance. Mohler K, Moen JM, Rogulina S, Rinehart...phosphopeptides in bacteria for proteome-wide identification of phosphorylation-dependent interactions. ...

...cell, and specifically targets the starter cell via the EnvA receptor. RABV is able to replicate in the ...which is not required for the transcription or replication of the genome within infected cells, but is required...present, the virus can spread from this cell to synaptically connected cells (meaning, it can spread to cells... cells that lack the G protein, the virus can replicate but the newly formed viruses are unable to infect...complementation, G-deleted rabies virus can spread to monosynaptically connected cells. To further refine the targeting... was modified so that it could only infect a genetically specified neuronal population. This is done using...leukosis virus. EnvA can direct virus infection specifically into cells that express the TVA viral receptor...

...loci seem to be associated with the disease and implicate a wide range of cellular processes. The vast majority...’s disease (PD) is a chronic and progressive neurological disorder that primarily affects movement. More... consists mainly of managing symptoms through medication and surgery. PD primarily involves the malfunction...available from Addgene, sortable by type of genetic modification and target species. You can also find our guide...Sun et al. Nat Methods. 2020 Oct 21. Glutamate indicators with improved activation kinetics and localization... transcription factors for sensory, motor, and cortical neuron differentiation. Held et al. Cell Rep. ...A multicolor atlas for whole-brain neuronal identification in C. elegans. Yemini et al. Cell. 2021 Jan...

...implementation letter DNA amplification Viral vector production Density centrifugation purification Addgene’s comprehensive...preps, which includes MTA facilitation, DNA amplification, and high-quality quality control measures such...preparations, we set both minimum standards and typical titers, though actual yields may vary by construct...construct. Size Quantity Total Volume Minimum Titer Typical Titer Small 10 × 100 µL aliquots 1 mL 4 × 10 12... with a minimum titer of 2 × 10 12 GC/mL and a typical titer of 7 × 10 12 GC/mL. For more details, see...remain subject to the terms and conditions of the applicable transfer plasmid. We will help you gain legal...providing scientist through an MTA. This process typically takes 1–4 days, depending on your institution....

...vector preparations were produced with the pUCmini-iCAP-PHP.eB plasmid (Addgene #103005) . Note on PHP.eB...vector preparations were produced with the pUCmini-iCAP-PHP.S plasmid (Addgene #103006) . Browse Available...vector preparations were produced with the pUCmini-iCAP-PHP.V1 plasmid (Addgene #127847) . Browse Available...vector preparations were produced with the pUCmini-iCAP-AAV.MaCPNS1 plasmid (Addgene #185136) . Browse Available...vector preparations were produced with the pUCmini-iCAP-AAV.MaCPNS2 plasmid (Addgene #185137) . Browse Available...vector preparations were produced with the pUCmini-iCAP-AAV.CAP-B10 plasmid (Addgene #175004) . Browse Available...vector preparations were produced with the pUCmini-iCAP-AAV.CAP-B22 plasmid (Addgene #175005) . Browse Available...