...validation and drug design Antibodies Addgene distributes ready-to-use recombinant antibodies made from plasmids... our trusted partner labs. Learn More ID Recombinant Antibody Reactivity Recommended Applications PI Don...Collections Search Plasmids Browse Plasmids Browse Antibodies Ordering and MTA Information Featured Collections...Golden Gate Assembly Standards Kit Cloning Eric Cantor Kit for using comprehensive end-joining ligation...junction sets for Golden Gate assembly. MoClo Toolkit Plant Expression Sylvestre Marillonnet 95 plasmids for...plasmids in our collection! These monoclonal antibodies are produced in-house and undergo application-specific...industry scientists. Please note this does not guarantee availability but lets us know what plasmids would...

...knockout )? Generate a specific mutant allele of a gene ( point mutant )? Increase or decrease expression...desired sequence, you may want to consider alternative Cas enzymes ( see Cas9 variants and PAM sequences ).... specific disease or process of interest. Do you want to: Generate complete and permanent loss of gene...Cas9-gRNA ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression. The table...may alter protein function when they occur in important protein domains. For gene editing experiments ... addition to off-target activity , it is also important to consider factors that maximize cleavage of ...assay (for NHEJ repaired DSBs): Provides a semi-quantitative readout of the percentage of alleles that have...

...PubMed PMID: 24379683 . When mutants gain new powers: news from the mutant p53 field. Brosh R, Rotter V...to activate transcription, and they also have a dominant negative effect on functional p53 through oligomerization... of p53’s pro-apoptotic effects is especially important to tumorigenesis. Inheriting only one functional...-of-function, expanding the mechanisms by which mutant p53 is known to function in cancer. p53 Pathway...: 24739573 . Uncovering the role of p53 splice variants in human malignancy: a clinical perspective. Surget...2003 Mar;21(3):313-20. PubMed PMID: 12619118 . p53 mutant mice that display early ageing-associated phenotypes...of p300/CBP in p53-dependent signal pathways. Avantaggiati ML, Ogryzko V, Gardner K, Giordano A, Levine...

...Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources...Design Tools CRISPR Blog Posts CRISPR/Cas nickase mutants introduce gRNA-targeted single-strand breaks in...created by wild type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands...specific edits via homology-directed repair (HDR). Want more information on the wide variety of Cas enzymes...expression in mammalian systems, bacteria, Drosophila, plants, and yeast. Mammalian ID Plasmid Gene/Insert Promoter...Insert Promoter Selectable Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker...

...first set of anti-CRISPR genes in bacteriophages infecting Pseudomonas aeruginosa . Anti-CRISPR genes ...Cpf1) Type IV (Csf1) Type VI (Cas13) Fighting Back: Anti-CRISPR Genes in Phage The CRISPR adaptive immune...complex. Recently, researchers have begun to use anti-CRISPRs as off-switches for CRISPR genomic editing...libraries can be used to identify genes that are important to a given phenotype. Current libraries are available...editing rates. Advances in this area include: Nickase mutant: cuts only one strand of DNA and can be used with... High fidelity Cas9s: Three high fidelity Cas9 variants , SpCas9-HF, eSpCas9, and HypaCas9 display very...to be used together. Researchers have engineered mutant Cas enzymes with alternative PAM sites . Cpf1 ,...

...cells and plate onto LB agar plates with the relevant antibiotic. Select multiple colonies, mini-prep each...expression of the relevant gene 7,8 . To evaluate the protein, perform an immunoblot using an antibody against ...joining (NHEJ) repair. Deletions have potential advantages as compared to single-site small indels given...complement oligo ( e.g., sgRNA-A in Table 4 ). The resultant oligos would be 25-mer oligos. However, if the...the reverse complement oligo. In this case, the resultant oligos would be 24-mer oligos ( Table 4 ). Design...Centrifuge PCR plate at 400 x g for 5 min and remove supernatant by flicking the PCR plate over a sink. Add 50...against the relevant protein 9 . References This protocol is excerpted from: Generation of Genomic Deletions...

... protein component of MMR) mutant to inhibit MMR PE5 - PE3 plus MLH1 mutant to inhibit MMR PEmax - optimized...QPM-sgR (pTaU3) Plant TaU3 nicking sgRNA Eps3I + NcoI No Caixia Gao 141081 pYPQ141D-peg Plant OsU3 pegRNA ... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources... for expression in mammalian systems, bacteria, plants, and Drosophila. Mammalian ID Plasmid Gene/Insert...Insert Promoter Selectable Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker...BbsI + PstI for pegRNAs No Xiao Wang 170132 pOsU3 Plant OsU3 pegRNA BsaI + HindIII No Caixia Gao 172716 ...

...Tissue Bank Antibodies (Link opens in a new window) N-terminal Anti-MECP2 (M7443) mouse antibody from Sigma...C-terminal Anti-MECP2 (M6818) mouse antibody from Sigma (Link opens in a new window) C-terminal Anti-MECP2 ... expressing wild-type MECP2 and half expressing mutant MECP2 . Skewing of these X inactivation ratios ...syndrome at the molecular level. They are also an important tool for developing therapeutic strategies to ...Anti-MECP2 (D4F3) rabbit antibody from Cell Signalling Technology Additional Resources (Link opens in a new window...

...Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair..., RNA, and DNA. The enCHIP system uses an anti-FLAG antibody to immunoprecipitate FLAG-tagged Cas9. In... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources... This technique creates diverse populations of mutants for directed evolution. ID Plasmid Gene/Insert ... Marker PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted single-strand breaks in...created by wild type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands...

...Reagents Protein phosphorylation is one of the most abundant forms of posttranslational modifications in cells...University changed the way researchers can explore important questions surrounding serine phosphorylation by...overall improved ease of use when expressing recombinant phosphoproteins. These attributes make rEcoli...in place of the popular SepOTSλ ( 68292 ). An important technical note: pSerOTS-C1* ( 188537 ) is a ROP...) is available from Addgene. This library is anticipated to be useful for laboratories performing phosphoproteomic...reference spectra or phosphopeptide standards for quantitative assessments. The mode #1 phosphosite library...188531 ; 188532 ; 188533 ; 188534 ; 188535 ). Importantly, phosphorylation positional isomers are allocated...

... For the most part, we will assume that you want to express a gene; however, we have a section...are studying a different genetic element or want to express shRNA. Choose by: Species-specific expression... modification Species-Specific Expression If you want to drive expression of your favorite gene, you will...be functional in your host organism. Host Relevant Promoters Representative Empty Backbones Mammalian... to easily identify your protein using an antibody against that epitope. This could allow you... your favorite gene even if you do not have an antibody against it. Another common scenario is fusing...sufficient for some experiments, scientists often want to create stable cell lines in which the plasmid...

... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources... base editors are available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements...fusions, modifying the linker region between Cas variant and deaminase to adjust the editing window, or ...systems create a wide spectrum of single-nucleotide variants (somatic hypermutation) in a wider editing window... to activation-induced cytidine deaminase (AID). Want more information on the wide variety of Cas enzymes... for expression in mammalian systems, bacteria, plants, yeast, and zebrafish. Mammalian ID Plasmid Gene...Insert Promoter Selectable Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker...

...Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair..., RNA, and DNA. The enCHIP system uses an anti-FLAG antibody to immunoprecipitate FLAG-tagged Cas9. Design... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources... Marker PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted single-strand breaks in...created by wild type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands...

...Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair..., RNA, and DNA. The enCHIP system uses an anti-FLAG antibody to immunoprecipitate FLAG-tagged Cas9. Design... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources...Promoter PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted single-strand breaks in...created by wild type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands...

...High-efficiency transduction of the mouse retina by tyrosine-mutant AAV serotype vectors. Mol Ther . 2009 Mar;17(3)...Mutated Adeno-Associated Virus Delivered to the Anterior Chamber Results in Efficient Transduction of Trabecular...Petrs-Silva, et al. 2011. Novel properties of tyrosine-mutant AAV2 vectors in the mouse retina. Mol Ther . Feb...mesenchymal stem cells into pluripotent stem cells by a mutant adeno-associated viral vector. Hum Gene Ther Methods...in Cynomolgus Macaques of rAAV2tYF-CB-hRS1, a Recombinant Adeno-Associated Virus Vector Expressing Retinoschisin...Sulfate Binding on Transduction of Retina by Recombinant Adeno-Associated Virus Vectors. J Virol . Mar... viral prep format. Please note this does not guarantee viral vector packaging service, but lets us know...

...ribosome and if the anticodon on the tRNA can bind to the mRNA (hence, the anticodon is complementary to... by tRNA variant that suppress the TAG nonsense codon, and tyrU gene replaced by tRNA variant inactivated... genes replaced by tRNA variant inactivated by the truncation of the anticodon loop). Expresses archaeal...aaRS M. jannaschii sulfotyrosine (sY) Bacterial, Plant TAG Chang Liu 85484 pDule-tfmF A65V S158A tri-fluoromethyl-phenylalanine...inactivated by the truncation of the anticodon loop). Expresses archaeal MjTyrRS/tRNA pair instead. For directed...knocked out, tyrTV and tyrU genes replaced by tRNA variant that suppress the TAG nonsense codon). Expresses...

...publications, focused on somatic variants found in lung adenocarcinoma as well as variants found across cancer types... for use in cancer signaling studies, including mutant, knockdown, and overexpression constructs. Each...involved in cancer development. These pathways are important to normal cell signaling and physiology, and, ...location, their pathway dependencies are extremely important for classification and treatment. On each pathway...Meyerson , and Root Labs containing wild-type and mutant alleles described in the initial Target Accelerator...

...CRISPR systems, the most abundant type found in bacteria and archaea. While abundant in nature, Class 1 systems... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources...double-strand break, either due to a boundary defined by anti-CRISPR proteins or running into other proteins bound... expression in mammalian systems, bacteria, and plants. Mammalian ID Plasmid Gene/Insert Promoter PI Publication... ID Plasmid Gene/Insert Promoter PI Publication Plant ID Plasmid Gene/Insert Promoter PI Publication Last...

...through our Packaged on Request service, Addgene guarantees a titer of at least 4 x 10 12 GC/mL for the AAV1... you know that we don’t expect to produce our guaranteed titer, and will give you the option to proceed...proceed with your order, or not, without a guaranteed titer. Any titer expectations communicated by email ...email will supersede the general guarantees above. We will have our expert viral vector production team carefully...our viral production does not yield the titer guaranteed here or by email, Addgene will contact you to...carried out with our materials, Addgene cannot guarantee the function of any viral vector. Addgene performs...

... infected by antibiotic resistant bacteria. Even when researchers develop new antibiotics, the onset of...there is no antibiotic strategy available to target only virulent or antibiotic resistant bacteria, and... fight against antibiotic resistance. Advantages of CRISPR over traditional antibiotics The potential ...CRISPR antimicrobials successfully resensitized a beta-lactam resistant population to the antibiotic, despite... the antimicrobial must reach essentially every cell or else small pools of antibiotic resistant cells...bacteria that are antibiotic resistant. Once these bacteria have been killed, antibiotics can be used to ...resensitize bacteria to antibiotics. A lysogenic phage infects antibiotic resistant bacteria, causing them...