...zinc finger (ZF) nuclease. Zinc fingers recognize triplets and the FokI nuclease operates as a dimer, cutting...) A TAL effector nuclease (TALEN). Similar in principle to the ZF nuclease, the components of the array...zinc fingers that recognize various nucleotide triplets. With some trial and error, ZF arrays are able...their limitations, however. Not every nucleotide triplet has a corresponding zinc finger, and interactions... targets are confined to sequences composed of triplets with corresponding zinc fingers, potential targetable... bp. TAL effectors have some restrictions (for example, the target must start with a T), but they still...Voytas/Bogdanove Golden Gate TALEN kit. In this example, the final array is composed of 18 TAL effector...

... we will use the example of CRISPR/Cas9 in mammalian cells, many of these principles apply to using CRISPR... process of interest. Do you want to: Generate complete and permanent loss of gene expression or function...major concern, Cas9-gRNA ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression...transgenic embryos Cas9-gRNA ribonucleoprotein (RNP) complexes Purified Cas9 protein and in vitro transcribed...transcribed gRNA are combined to form a Cas9-gRNA complex, which is delivered to cells using cationic lipids Transient... will depend on your specific application. For example: To activate or repress a target gene using dCas9...nucleotides within the selected target sequence. For example, gRNA targeting sequences containing a G nucleotide...

...sfGFP David Spray 54959 mApple-VE-Cadherin-N-10 Tight Junctions VE-Cadherin mApple Michael Davidson 55165... that has a different emission wavelength. For example, if the marker protein is fused to GFP, you may...with their own unique function. The nucleus, for example, houses the cell’s genetic material. The mitochondria...Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein Collection...Ciliary Base Chibby1 GFP Ken-Ichi Takemaru 89770 pLenti-EGFP-hChibby1 Mother Centrioles / Ciliary Base ...Base Chibby1 EGFP Ken-Ichi Takemaru 118084 pLenti-EB1-EGFP Microtubules EB1 EGFP Ken-Ichi Takemaru 122871...

... lot. At Addgene, AAV particles are titered by droplet digital PCR (ddPCR) using primers and probes targeting...Vector Core is (as of April 2016) also performed by droplet digital PCR (ddPCR). Quality Control Addgene ensures...specific QC experiments were performed on your lot, please contact us . Full sequencing of the Viral Genome...of the viral capsid proteins VP1, VP2 and VP3. Samples of viral preparations are subjected to polyacrylamide...proteins as a fraction of total protein present in the sample is also determined and used to determine purity...shown relative to the total protein present in the sample. Sterility Viral vector preparations are added ...Lentiviral vectors are titered using a lentiviral droplet digital PCR (ddPCR) titration protocol (modified...

...Nature Protocols 2013). For cloning information, please view: Zhang Lab General Cloning Protocol 237.2 ...SpCas9 Plasmids The CRISPR/Cas system can be implemented in mammalian cells by co-expressing the S. pyogenes...Activation Mediator (SAM) is an engineered protein complex for the transcriptional activation of endogenous...gain-of-function screening. For more information, please visit the Addgene SAM library page . Targeted gene... : Expresses the MS2-P65-HSF1 activator helper complex with 2A GFP 61424 : sgRNA cloning backbone with...vector encoding the MS2-P65-HSF1 activator helper complex with a 2A Hygro resistance marker 61427 : lentiviral... In these mice the CRISPR-Cas9 system can be implemented by delivering Cre and sgRNA to a Cre-dependent...

...and protocols showing examples of AAV strategies for intersectional control. Please see our list of suggested...publicly, but please only share materials that you are comfortable and willing to share. Please only share...information. Experimental protocols or schematics. Examples of research materials to share: In vivo data: ...Detailed protocol: AAV Production in HEK293 Cells Please share materials by uploading to your Dropbox link...contributions and your commitment to open science! Please see the meeting preparation section for how to ...

...cilia, centrosomes, the plasma membrane, lipid droplets, the nucleus, and mitochondria. The activity of...ARF GTPase family, information on each gene, and examples of plasmid types available for each gene. Click...gene name for a full list of plasmids available. Please note that some genes may not yet have plasmids ...request. Family Gene Entrez Gene ID Length (aa) Example Expression Systems GTPase Arf1 375 181 Bacterial... and GEF family, information on each gene, and examples of plasmid types available for each gene. Click...request. Family Gene Entrez Gene ID Length (aa) Example Expression Systems GAP Arfgap1 55738 414 Bacterial...

.... With its rapid development (embryogenesis is complete in ~3 days), short generation time (~3 months)...Highlights of Zebrafish Tools in our Collection CRISPR Please refer to Addgene's CRISPR Guide for a general introduction...expressing Cas9 and empty gRNA backbones. A few examples of useful zebrafish CRISPR resources can be found...visualize and track clones of cells over time. Examples of useful tools for studying cells that share ... Alex Schier Lab Other Tools of Interest A few examples of popular tools to use in zebrafish, ranging ...for studying diseases, are listed below: Lipid droplet reporters - Richard White Lab. An imaging platform...dynamics in vivo. Bimolecular fluorescence complementation (BiFC) tools - Marco Morsch Lab. A vertebrate...

...validated gRNA, based on the Zhang lab's PX458 . Multiple gRNA plasmids may be used. The HDR (homology directed...endogenous genes for the isolation of native protein complexes. This vector serves as a backbone to clone the...your experiments can be found below and in the supplemental materials from the Doyon Lab’s publication: ...targeting the PITCh donor plasmid. Note that the principle of the PITCh system is different from the conventional...Degron Tagging Masato Kanemaki's lab has developed a simple and scalable CRISPR/Cas-based method to tag endogenous...nuclear and cytoplasmic proteins, which can then be depleted very rapidly after the addition of auxin to the...and can be scaled up for systematic editing of multiple genes. Plasmids can be found associated with the...

...cloning grade DNA, please let us know using the Cloning Grade DNA Suggestion Form . Please note that this ...µl (± 5ng/µl) Buffer 10 mM Tris-Cl, pH 8.5 Price Please log in to your account and check the individual..., to order more than 2 µg of a single plasmid, multiple orders would need to be placed through our website...website. Every order will need to be completed independently including the check out and will require separate...I find cloning resources or scientific support? Please see our Addgene Protocols for resources you may...

...methylated DNA and the recruitment of the NCoR complex for proper nervous system function. While there...RSRT cell line collection (186.8 KB) . To order, please email [email protected] The following cell lines are...information on generating iPSCs using plasmids, please see Addgene's Plasmids for Stem Cell Research page...precise editing of mutations in the MECP2 gene. Please refer to Addgene's CRISPR Guide for a general introduction...expressing Cas9 and empty gRNA backbones. A few examples of additional CRISPR resources can be found below...cell line repository (Link opens in a new window) Sampled - one of the world's largest biorepositories (Link...your plasmids! To get started with your deposit, please click here ....

...total protein content of the sample is determined. Only antibody samples with >90% purity are distributed... webpage. To learn more about these antibodies, please visit the Institute for Protein Innovation Antibody...using a NanoDrop spectrophotometer. If needed, the sample is adjusted by diluting or concentrating to a final...Coomassie staining. For Coomassie staining, the sample is separated via a denaturing SDS-PAGE gel alongside...specific QC experiments were performed on your lot, please email [email protected] . Immunocytochemistry (ICC...

...imaging techniques, including that it is easy to implement due to the simplicity of gRNA design, programmable...for different genomic loci, capable of detecting multiple genomic loci, and compatible with live cell imaging...copies of labeled proteins to the given region. For example, chromosome-specific repetitive loci can be efficiently...visualized in living cells using a single gRNA that has multiple targeted sequences in close proximity. A non-repetitive...genomic locus can also be labeled by co-delivering multiple gRNAs that tile the locus. Chromosome painting...

...pAAV-synP-FLEX-splitTVA-EGFP-B19G Can be used to complement deletion-mutant rabies virus AAV1 Wickersham ...pAAV-syn-FLEX-splitTVA-EGFP-tTA These AAV together can be used to complement deletion-mutant rabies virus AAV1 Wickersham ...lacks the ability to infect neighboring cells. For example, G-deleted rabies lacks the viral glycoprotein...infect any neighboring cells. Thus, through complementation, G-deleted rabies virus can spread to monosynaptically...would like to see available in viral prep format. Please note this does not guarantee viral vector packaging...

... seed selection and multiplex CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants...biology - deposit plant plasmids with Addgene! CRISPR Please refer to Addgene's CRISPR Guide for a general introduction...variants of Cas9 and empty gRNA backbones. A few examples of useful plant CRISPR resources include: A Multipurpose...CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants - Yao-Guang...

...far more complex than using CRISPR to modify a single genomic locus. Before ordering, please review the...consist of thousands of plasmids, each containing multiple gRNAs for each target gene. In a CRISPR screening...Root 2nd 2 40,710 Inzolia Human CRISPR/Cas12a Multiplex Knockout 209551 209552 Knockout Human Doench 3rd...117725 Knockout Human Vakoc 3rd 6 3,051 Human Lipid Droplet and Metabolism Library 191535 Knockout Human Olzmann...available items found to match all requested criteria. Please consider using fewer filtering options. Browse ...

... browse the gene list below. For proteins with multiple subunits or isoforms, individual links to each...available plasmids for the gene. For components with multiple subunits or isoforms, individual links to each...EXOC2 EXOC3 EXOC4 EXOC5 EXOC6 EXOC7 EXOC8 Exocyst complex component FGFR FGFR1 FGFR2 FGFR3 FGFR4 Fibroblast... cis/trans isomerase, NIMA-interacting 1 PLXNB1 Plexin B1 PPP1CA Protein phosphatase 1 catalytic subunit...B1 RPTOR Regulatory associated protein of MTOR, complex 1 SAV1 Salvador family WW domain containing protein...

...movement. More than ten million people worldwide are living with PD, mostly people over the age of 60. There...caused by abnormal expansion of a repeating CAG triplet series within the huntingtin (HTT) gene. An individual...variation and lifestyle or environmental factors. For example, variations of Apolipoprotein E (APOE), such as...collection of neurological mouse models including complex and monogenic diseases. Alzforum (Link opens in...

...PGK Retroviral Lowe 33344 pLEW100cre-EP1-6G Cre T. brucei Cross 33345 pLEW100cre-EP1 Cre Trypanosoma brucei...blog post . Based on these Cre-lox recombination principles, scientists have developed constructs to activate...pSkipFlox split Cre Pfcrt P. falxiparum Spielmann 86641 pLenti-hSynapsin-Cre-WPRE Cre hSyn Lentiviral Wang 86794...donor/transfer vector Cbh AAV Eisenhoffer 131786 pLenti-CreLite CreLite system components, PhyBdeltaCreC...in the indelible marking of mouse neurons with multiple distinct colors Mammalian Sanes See article Multibow... to facilitate Cas9 mediated genome editing in multiple tissues. FLP/FRT: Alternative recombinase system...

...cut S. pyogenes 26028531 Huangfu OCT4 H. sapiens multiple, see article 69537 activate S. pyogenes 26352799... pyogenes 25336740 Christiaen EGFP A. victoria multiple, see article 60071 dCas9-FokI S. pyogenes 24770325...23817069 Calarco Kras, p53, and Lkb1 M. musculus multiple, see article 60224 cut S. pyogenes 25263330 Zhang.... pyogenes 23929340 Kamoun PDS1 N. benthamiana multiple, see article 49772 cut S. pyogenes 24112467 Kamoun...AGCACTGTTGCCGGCCGAGG 79006 cut S. pyogenes 26944679 Shaw Please refer to the primary article to confirm details...add validated gRNA sequences to our datatable, please download the following spreadsheet, fill out as...