...or MCP tagged plasmids for visualization of proteins inside cells....-, ACP-, and MCP-tags for visualization of proteins inside living or fixed cells. The plasmids associated... labeling cell-surface proteins, and are useful for labeling secreted proteins with disulfide bridges ...features: Allows for visualization of multiple proteins simultaneously with dual labeling when used with...live or fixed cells, and for internal or surface proteins Single constructs are compatible with multiple... and MCP-tags are used exclusively for surface proteins and use enzymatic binding to attach substrate ...suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag is identical in function to...

...correct insert (gRNA) in pX330 / pX335 with BbsI/AgeI double digestion. As a successful insertion will destroy... have been published with thorough step-wise instructions on designing and using Cas9-based genome engineering...vector (like PX330). For large changes (>100bp insertions or deletions), we typically use plasmid donor...homology arms on each side flanking your desired insertion or mutation. Each arms is around 800bp. For transfection...CRISPR/Cas9 system to create specific mutations or insertions by Homologous Recombination (HR), what is the...purified long oligos. For large changes (>100bp insertions or deletions), the Zhang lab typically uses a...homology arms on each side flanking your desired insertion or mutation. Each arm is around 800bp. For transfection...

...Mullineaux Insulin Sensor for Imaging Insulin Secretion A Ratiometric Sensor for Imaging Insulin Secretion... Collections Fluorescent Proteins Fluorescent Biosensors Fluorescent Proteins: Biosensors Metal Ions Small...Service: AAV Biosensors Fluorescent biosensors are proteins designed so that their fluorescent properties ...transfer (FRET) efficiency of a pair of fluorescent proteins. Genetically encoded biosensors can be introduced...Constitutive or Cre-dependent) Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature. 2013 Jul...nested reporters of green- and orange-emitting proteins Ratiometric Matryoshka biosensors from a nested...cassette of green- and orange-emitting fluorescent proteins. Nat Commun. 2017 Sep 5;8(1):431. Wolf Frommer...

... EF1a-tTA-2 Contains Gateway L1L3 sites and EF1α promoter driving tTA; contains insulator sequence; plasmid...Blasticidin selection. Please NOTE, this vector contains an insert (MAT2A) which would need to be replaced by...Backbones Find a construct that will allow you to insert and induce your gene of interest. ID Plasmid Description...format: PGK-rtTA-2A-puro; see article for tagged insert options rtTA On Root 11651 pLVUT-tTR-KRAB Lentiviral...bidirectional tet-responsive promoter (Pbi); Pbi contains a TRE between two minimal CMV and is silent in...third-generation rtTA On Elledge 19407 pTREtight2 Promoter contains a modified TRE that is silent in absence of binding...

...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter PI...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast ID Plasmid Gene/Insert Promoter Selectable...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Xenopus ID Plasmid Gene/Insert Promoter Selectable...joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often...repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs...Marker PI Publication Parasites ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Do you...

... Plasmid Collections Fluorescent Proteins FRET Fluorescent Protein Guide: FRET More Fluorescent...fluorescent. A variety of organic dyes and fluorescent proteins have been developed for use as donor and acceptor...intramolecular or unimolecular FRET). Fluorescent proteins can be used as donor and/or acceptor molecules...suitable for creating individual fluorescently tagged proteins to study protein-protein interactions and a series...laboratory. Empty Vectors Encoding Fluorescent Proteins for FRET The following plasmids can be used to...brightness standard consisting of two Venus fluorescent proteins connected via a 5 amino acid linker V17V Anisotropy...brightness standard consisting of two Venus fluorescent proteins connected via a 17 amino acid linker V32V Anisotropy...

... System Contains 336 pFusX clones allowing for Golden Gate assembly of 15-16 RVD TALEN inserts in one ...plasmids of pre-assembled trimer and tetramer RVD domains designed for rapid TALEN assembly. Kiran Musunuru...message. Both vectors utilize homodimeric FokI domains. 38143 pC-GoldyTALEN Golden Gate TALEN 2.0 36033...sequences within core promoter regions. pTAL5-BB contains the GAL1 promoter, placing TALORs built into this...under galactose-inducible expression. pTAL6-BB contains the TEF1 promoter, giving constitutive expression...mutation induction when using TALENs. The FokI domains (DD, RR) used are obligate heterodimers that require...require cloning of left and right TALEN monomer proteins into opposite vectors. 37276 pCS2TAL3-RR 48636...

...reverse transcriptase (RT) is capable of installing targeted insertions, deletions, and all possible base-to-base... been designed for use in Drosophila and other insects. CRISPR... been designed for use in Drosophila and other insects. Cut Fully functional CRISPR/Cas enzymes will introduce...joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often...repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs...much lower efficiency than NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Nick CRISPR/Cas nickase...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type...

...) that contains dCas9 fused to N 51025 pSLQ1661-sgMUC4-E3(F+E) Lentiviral vector that contains sgRNA targeting...CRISPRi is a system that can regulate a genome instead of modifying a genome. 1. Two-plasmid CRISPRi system...knockdown The first plasmid ( pdCas9_bacteria ) contains an anhydrotetracycline (aTc)-inducible dcas9 gene...resistance. The second plasmid ( pgRNA_bacteria ) contains a constitutive sgRNA expression cassette on a ...knockdown The first plasmid ( pdCas9_humanized ) contains a human codon optimized dcas9 gene under the control...promoter. The second plasmid ( pgRNA_humanized ) contains a murine U6 promoter controlled sgRNA cassette...sequences in the genome. The pgRNA_humanized also contains a CMV-puro-t2A-mCherry expression cassette, useful...

... TAG Edward Lemke Browse Strains The table below highlights bacteria strains that have been modified to...Addgene's guide to plasmids and strains useful for expanding the genetic code in bacteria and mammalian...Rinehart Lab reagents tRNA/Synthetase Pairs Bacterial Strains Codon Modified Constructs The genetic code for ...genetic code can help us understand and evolve proteins for various purposes from therapeutics to biopolymers...phosphoserine GCE. For expressing phosphoserine-containing proteins using genetic code expansion without buildup of...DE3) ΔserC serC gene knocked out; for expressing proteins containing site-specific non-hydrolyzable phosphoserine...and trpT genes). Expresses yeast TrpRS/tRNA pair instead. For directed evolution of the liberated EcTrpRS...

...reverse transcriptase (RT) is capable of installing targeted insertions, deletions, and all possible base-to-base...joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often...repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs...much lower efficiency than NHEJ. ID Plasmid Gene/Insert Selectable Marker PI Publication Base Edit Catalytically...the opposite strand) mutations. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick ...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type...homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Prime...

...joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often...repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs...much lower efficiency than NHEJ. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Base ...the opposite strand) mutations. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick ...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type...homology-directed repair (HDR). ID Plasmid Description Gene/Insert Promoter Selectable Marker PI Publication Activate...activator to your specific locus. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Interfere...

...Plasmid Collections Fluorescent Proteins Organelle Markers Fluorescent Proteins: Subcellular Localization More...as these well-characterized proteins. General Structures The cell contains a variety of organelles and...role in secreting proteins from the cell to the extracellular environment. Proteins are first translated...Plasmids encoding fluorescent proteins tagged with genes or peptides with known subcellular localization...Fluorescent Protein Resources: Empty Backbones Allen Institute for Cell Science Plasmid Collection FRET Biosensors...has assembled a collection of fluorescent fusion proteins that are targeted to various organelles or subcellular...FAP; mCerulean3 Marcel Bruchez 45944 pTDpelB-C_sfYFPTwinStrep Periplasmic space PelB signal sequence YFP...

...Genetics 2014 Allen Institute for Cell Science Plasmid Collection The Allen Institute for Cell Science has...CRISPR system has been adapted to allow tagging of proteins expressed from their natural chromosomal context...add a tag (currently using FLAG) to endogenous proteins. This CRISPR/Cas plasmid-system consists of two...2A-PuroR cassette to the C-terminus of endogenous proteins. The PITCh system depends on MMEJ (microhomology-mediated...end-joining), an alternative DSB repair pathway, instead of HDR, which makes it easy to construct a donor...'s lab for building CRISPR targeting vectors to insert genetic tags in the C. elegans genome. The SapTrap...template for an individual tagging event. The kit contains 26 plasmids; 21 of the plasmids are for use in...

...our repository that express fluorescent proteins. ID Name Insert Antibiotic Resistance Purpose PI 17446...Plasmid Pooled Libraries . ID Name Description Genes/Insert Selection PI Human knockout pooled libraries 73178...with 1000 non-targeting controls. This backbone contains SpCas9. SpCas9 and 76,441 unique sgRNAs targeting...interrogate the genome in a variety of ways. Cas9 proteins can be guided to specific genomic sites by an ... ) or single-stranded breaks ( nicks ). ID Name Insert Function Antibiotic Resistance Activity PI 52962...and used as a transcriptional activator . ID Name Insert Antibiotic Resistance Activity PI 61422 dCAS9-VP64...barcoding library. Version 1 of the Celltag library contains 19973 barcodes to combinatorially index cells ...

...PubMed . Direct identification of insulator components by insertional chromatin immunoprecipitation. Fujita... Locus-specific ChIP technologies consist of insertional chromatin immunoprecipitation (iChIP) and engineered...with engineered DNA-binding molecules such as TAL proteins and the CRISPR system consisting of a catalytically...s13104-018-3486-3. PubMed . Promoter-associated proteins of EPAS1 identified by enChIP-MS - A putative ...enChIP technology using recombinant CRISPR ribonucleoproteins. Fujita T, Yuno M, Fujii H. Genes Cells. ...system using recombinant exogenous DNA-binding proteins.Fujita T, Fujii H. BMC Mol Biol. 2014 Nov 27;15:26.../s12867-014-0026-0. PubMed . Identification of proteins associated with an IFNgamma-responsive promoter...

...(deletions or small insertions) or (B) span the genome-insert boundary (insertions only). The PCR product...dual) gRNA targeting 5′ exon or essential protein domains High-fidelity Cas enzymes increase specificity.... gene, such as generating a point mutation or inserting a tag Cas9 (or Cas9 nickase); Base editor Single...using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer vector and used to generate...target exons that code for known essential protein domains. The benefit of this approach is that even non-... function when they occur in important protein domains. For gene editing experiments using HDR, it is ...targeting oligos are synthesized, annealed, and inserted into plasmids containing the gRNA scaffold using...

...to boost the research focused on Solute carrier proteins (SLCs). Browse the SLC plasmid collection....Plasmids Pooled Libraries Feedback Solute carrier proteins (SLCs) are the largest family of transporters ...associations. Plasmids This plasmid collection contains codon-optimized ORF sequences for human SLCs in...Gateway-compatible pDEST vectors. ID Plasmid Gene/Insert Industry PI Pooled Libraries CRISPR and overexpression... libraries target the solute carrier family of proteins. Library ID PI Description Human SLC Activation... CRISPR activation library targeting human SLC proteins. Human SLC Knockout Library 132552 Superti-Furga...Superti-Furga CRISPR knockout library targeting human SLC proteins. Mouse SLC Knockout Library 141436 Superti-Furga...

...of interest, a medium copy RSF1030 origins or a low copy p15a origins should be used. Find further details...system to make singly or multiply phosphorylated proteins (Pirman et al., Nat Comm 2015). Improved phosphoprotein...improved ease of use when expressing recombinant phosphoproteins. These attributes make rEcoli XpS ( 192872 ...other plasmids containing high copy ColE1/PUC-like origins. Therefore, when choosing plasmids to express the...sequences corresponding to every previously observed instance of human serine phosphorylation ( Rinehart Human...phosphosite library with four different phosphobinding domains (14-3-3β, 14-3-3σ, the WW2 domain of NEDD4, and...68307 ) to generate the phosphosites with serine instead of phosphoserine (i.e. a “phosphorylatable” phosphosite...

...nerve cell function within the cell. Parkinson's Disease Parkinson’s disease (PD) is a chronic and progressive...More Fluorescent Proteins Find information on Addgene's collections of fluorescent proteins, including Markers...Differentiation Factors Antibodies Fluorescent Proteins Other Resources Neurodegenerative Disorders Neurodegenerative...neurodegenerative diseases include Alzheimer’s, Parkinson’s, ALS, and Huntington’s Disease. These diseases...with the mutated HTT gene will have huntingtin proteins with abnormally long polyglutamine sequences. ...with concomitant expression of aggregation-prone proteins with the iPSC toolbox . Lam et al. bioRxiv. 2022...A foundation dedicated to finding a cure for Parkinson's disease through an aggressively funded research...