... 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...

...) 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...

...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...a much lower efficiency than NHEJ. Plasmid Gene/Insert Selectable Marker PI Publication Base Edit Catalytically...on the opposite strand) mutations. 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...via homology-directed repair (HDR). 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...a much lower efficiency than NHEJ. Plasmid Gene/Insert Promoter Selectable Marker PI Publication Base ...on the opposite strand) mutations. 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. Plasmid Gene/Insert Promoter Selectable Marker PI Publication Interfere...

...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...

...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...

...sgRNA without PBS. It contains extra sequences for BsaI digestion for insertion of PBS 71899 pAC1399-pX-sgRNA...These PUF-binding sites recruit PUF (Pumilio/FBF) domains that are fused to an effector, like those for epigenetic...alternate strategy to techniques that fuse effector domains to dCas9. PUF can be programmed to recognize virtually...loci. It can also potentially recruit different proteins on a single sgRNA allowing the study of protein...the future, modules tethering different fusion proteins can be assigned a unique PUF/PBS pairing. The ...4xNLS_PUFa_2xNLS NLSPUFa pCR8 gateway donor vector for insertion of N-terminal domain (SgrAI or AgeI with ClaI)...4xNLS_PUFb_2xNLS NLSPUFb pCR8 gateway donor vector for insertion of N-terminal domain (SgrAI or AgeI with ClaI)...

...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...

...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...Foundation is dedicated to finding a cure for Parkinson's disease through an aggressively funded research...

...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 ...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...

...use. Negative Result 1: Both Strains Grow Assuming the appropriate strains were streaked on the appropriate...your strains are no longer viable. You can check for this possibility by streaking out both strains on ...slightly. Even so, you should always use thermally insulated gloves when removing anything from the autoclave...appropriate plates, then if both strains grow, it’s possible that: The antibiotic broke down. You forgot to...concentration for selection. Negative Result 2: Neither Strains Grows If neither strain grows, it's possible that...ve used the wrong antibiotic, or confused your strains....

...important plasmid features (such as the gene/insert, fusion proteins, point mutations, deletions, etc.) involves...backbone and can help you verify the ends of the insert. Longer genes may require internal primers that...that anneal to a specific region of the insert. You can find Addgene's sequencing results by clicking on... to verify tags, mutations and a portion of the insert, but we do not sequence the entire plasmid. Addgene...and make sure they are justifiable peaks. For instance, in the trace file below, you can see that just...Sometimes an “N” is the result of an erroneous insertion by the sequencing reaction....

...ID Plasmid Description Gene/Insert PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids... vectors for tag insertion using CRISPR/Cas9 and the selection of C. elegans strains. CRISPR plasmids ...includes CRISPR, RNAi, empty backbones, fluorescent proteins, and Cre expression plasmids. Large collections...briggsae , etc.) gene or sequence: ID Plasmid Gene/Insert PI Don’t see your gene listed? Know of some great...elegans genome engineering - Bob Goldstein Lab. Contains a set of plasmids for building homologous repair...Transgenesis Kit - Erik Jorgensen Lab. This kit contains 29 plasmids used to generate transgenic nematodes...Caenorhabditis Genetics Center (CGC) - The CGC collects, maintains, and distributes stocks of C. elegans. Silencing...

...Includes many N- and C-terminal fluorescent proteins Insect Sutherland Lab Plasmids - For multicistronic...different fluorescent tags for you to create fusion proteins with your gene of interest. Special... Monomer not a Monomer? Background Fluorescent proteins are genetically encoded tools that are used extensively...have engineered numerous GFP-variants and non-GFP proteins that result in a diverse set of colors. Addgene...different fluorescent tags for you to create fusion proteins with your gene of interest. These are organized... at high concentration, so this is noted here. Proteins that are derived from GFP and contain the A206K...information on the properties of many of these proteins, see: Cranfill et al. , Nature Methods 2016 Jul...

...to boost the research focused on Solute carrier proteins (SLCs). Browse the SLC plasmid collection.... Collection NeuroMab Collection 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 Pooled Libraries... 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...

...staining cells in culture with a primary antibody against a target protein and a fluorescent secondary antibody...labeling cells in culture with a primary antibody against a target protein and a fluorescent secondary antibody...need to be optimized for different cells, target proteins, etc. Sharing speeds science. We believe that ... Before Starting Refer to the manufacturer's instructions for additional information specific to your ...Paraformaldehyde fixation works well for many target proteins, it may not be the best fixation method for all...Remove the paraformaldehyde and follow your institution's laboratory safety guidelines for disposing of...between antibodies. Review the manufacturer's instructions before starting your experiment and consider...

...into the host genome, it has the potential for insertional mutagensis. The 2nd and 3rd generation lentiviral...Type Description PI 12253 pRSV-Rev 3rd Packaging Contains Rev; can be used with 3rd generation transfer ...Rawlings , Scharenberg 12251 pMDLg/pRRE 3rd Packaging Contains Gag and Pol; can be used with 3rd generation transfer... encoding Rev Trono 12260 psPAX2 2nd Packaging Contains Gag, Pol, Rev, and Tat; can be used with 2nd and...plasmids Trono 12263 pCMV delta R8.2 2nd Packaging Contains Gag, Pol, Rev, and Tat; can be used with 2nd and...plasmid Weinberg 8455 pCMV-dR8.2 dvpr 2nd Packaging Contains Gag, Pol, Rev, Tat, Vif, and Nef; can be used ...envelope plasmid Kasparov 22500 pNHP 2nd Packaging Contains gag, pol, tat, rev Kasparov 1732 pCPRDEnv 2nd ...

... Carrier Proteins (SLCs) from the RESOLUTE collection AAV Packaging from Addgene Parkinson's Research ...SARS-COV-2, ACE2, and CRISPR tools Fluorescent Proteins from the Michael Davidson collection Protein Labeling...available to industry scientists. ID Plasmid Gene/Insert PI Antibodies Addgene distributes ready-to-use ...agreement between the recipient and the depositing institution(s). It governs the transfer of biological materials...distribute plasmids on behalf of the depositing institutions. These agreements indicate the language which... by an authorized signatory at the recipient institution. An authorized signatory is someone who can execute...execute legal documents on behalf of the recipient institution. Once the MTA has been signed, it can be submitted...

...Bertozzi Nicolas Buchler George Church Cynthia Collins James Collins Tom Ellis Drew Endy Tobias Erb Susan Golden...environmental sensing. Strains Browse Addgene's collection of engineered bacterial strains, including plasmid-strain...terminators, repressors, activators, and more. Also contains pre-assembled genetic circuits such as logic gates...Autoregulation and Yeast-Mammalian Circuit Transfer C. Collins Lab Quorum Sensing Ellis Lab GeneGuard Endy Lab...Addgene Tools and Resources CRISPR Fluorescent Proteins FREQ-Seq Kit FX Cloning Kit GreenGate Cloning ...ethical synthetic biology JBEI - Joint Bio-Energy Institute Synberc - Synthetic Biology Engineering Research...