...Alpha-tubulin Microtubules 101786 ST6GAL1-mEGFP AICSDP-26 mEGFP Sialyltransferase 1 Golgi 109122 NPM1-mEGFP AICSDP...Institute ID Tag Protein Structure 87420 PXN-EGFP AICSDP-1 EGFP Paxillin Matrix Adhesions 87421 TUBA1B-mEGFP ...TJP1-mEGFP AICSDP-23 mEGFP Tight junction protein ZO-1 Tight junctions 91565 AAVS1-mEGFP AICSDP-35 mEGFP ...Centrioles 101782 LAMP1-mEGFP AICSDP-19 mEGFP LAMP-1 Lysosome 101783 MAP1LC3B-mEGFP AICSDP-25 mEGFP Autophagy-related... HIST1H2BJ-mEGFP AICSDP-52 mEGFP Histone H2B type 1-J Histones 109119 CTNNB1-mEGFP AICSDP-47 mEGFP Beta-catenin...

... the 3′ end (see Figure). Mammalian System: Table 1 We provide a collection of reagents for custom CRISPR...transcriptional activator. Streptococcus thermophilus #1 (ST1): Almost as small as NM, ST1 consistently yields...Description 48669 M-ST1cas Mammalian S. thermophilus #1 Cas9 expression, human optimized 48670 M-NMcas Mammalian...spectinomycin 48647 DS-ST1cas Bacterial S. thermophilus #1 Cas9 (ST1) + tracrRNA expression, cloDF13/spectinomycin...budding yeast) from the TEF1 promoter 43804 p415-GalL-Cas9-CYC1t A human codon-optimized Cas9 for expression...expression in S. cerevisiae (budding yeast) from the GalL promoter 43803 p426-SNR52p-gRNA.CAN1.Y-SUP4t A gRNA...

... pU6-sgGAL4-1 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GAL4 UAS promoter... regulate a genome instead of modifying a genome. 1. Two-plasmid CRISPRi system for bacterial gene knockdown... pU6-sgGAL4-4 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GAL4 UAS promoter...

...Heidenreich 1, Banerjee A, Habib N, Li Y, Trombetta J, Sur M, Zhang F. Nat Biotechnol . 2015 Jan;33(1):102-6...SpCas9 alone without sgRNA. Full references are below. 1. SpCas9 (or SpCas9n, D10A nickase) + single guide ...screens. The libraries are available in two formats: 1 vector system - lentiCRISPR - sgRNA and SpCas9 together...):186-91. doi: 10.1038/nature14299. Epub 2015 Apr 1. PubMed . Return to AAV In vivo plasmids AAV Plasmids...codon-optimized SaCas9 driven by either the ubiquitous cytomegalovirus (CMV, PX601 [#61591] ) or liver-specific tyroxine...

...Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert pLKO.1-puro U6 sgRNA BfuAI stuffer 50920 Mammalian/Lentiviral...Worm HindIII none S. pyogenes de Bono sgRNA with rpr-1 promoter 48961 Worm EcoRI none S. pyogenes de Bono...Zebrafish BseRI none S. pyogenes Zon pCRISPomyces-1 61736 Bacteria BbsI yes, cut S. pyogenes Apramycin...see plasmid page yes, cut S. pyogenes BFP Kuhn pLKO.1-puro U6 sgRNA BfuAI large stuffer 52628 Mammalian/...Puro Zoldos pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110) 71707 Mammalian hU6 yes, cut S. pyogenes Draetta...expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent promoters. sgRNAs...for the expression of two gRNAs from Drosophila U6:1 and U6:3 promoters. Bullock pCFD5 Drosophila Utilizes...

... your account. We will notify you by email within 1 business day regarding your account verification. ... transfers to for-profit entities. The MTA is a legal agreement between the recipient and the depositing...authorized signatory is someone who can execute legal documents on behalf of the recipient institution...

...feedback on this service. AAV Service Details Volume 1-1.5 mL in 100 µL aliquots Minimum Titer 5E12 GC/mL... feasible. Please note that we cannot guarantee legal approval to prepare a plasmid as viral particles...

...interactions. Sun Y, Gallagher-Jones M, Barker C, Wright GJ. Analytical Biochem. 2012 May 1;424(1):45-53....

... your experiment. Libraries may be available in a 1-plasmid system, in which Cas9 is included on the gRNA-containing...sequence for Staphylococcus aureus Cas9 (SaCas9) is ∼1 kb shorter than SpCas9, SaCas9 can be efficiently ...sequences containing a G nucleotide at position 20 (1 bp upstream of the PAM) may be more efficacious than...population. Some cells may be wild type due to either (1) a lack of gRNA and/or Cas9 expression or (2) a lack...Giovannangeli C, Concordet JP. Nat Commun . Mar 19;9(1):1133. PMID: 29556040 Directed evolution of CRISPR-Cas9...Jung I, Kim D, Kim S, Kim JS. Nat Commun . Aug 6;9(1):3048. PMID: 30082838 Engineered anti-CRISPR proteins...DE, Piccioni F, Doench JG. Nat Commun . Dec 21;9(1):5416. PMID: 30575746 Synthetic CRISPR-Cas gene activators...

...using centrifugal columns. The final formulation buffer is phosphate-buffered saline containing 1 mM sodium...

...disadvantages: Table 1 (Joglekar et al., 2017)  Table 1 (Gutierrez-Guerrero et al., 2020) Table 1 (Cronin et al...polyA tail.   Figure 1: Lentiviral production uses three plasmids: (1) The transfer plasmid, (2)... specific purposes as described previously (Table 1, Gutierrez-Guerrero, et al, 2020). Because retrovirus...virus in cell lines expressing galactosyl(alpha1-3)galactosyl (alphaGal) sugars were less stable than ...and spumaviruses to human serum by galactosyl(alpha1-3)galactosylation. Journal of virology 71:6174–6178...express these sugars. (Takeuchi et al., 1997). The alphaGal sugars end up in the envelope and are targets ...targets for complement-based killing by anti-alphaGal antibodies. The VSV-G envelope protein is inactivated ...

...intracellular targets. Nature Communications, 7(1), Article 1. https://doi.org/10.1038/ncomms10778 Erdmann...molecules in vivo. Nature Biotechnology, 21(1), Article 1. https://doi.org/10.1038/nbt765 Lavis, L. (2016... envelope design. Nature Communications, 14(1), Article 1. https://doi.org/10.1038/s41467-023-40788-8 ... capping protein. Nature Communications, 14(1), Article 1. https://doi.org/10.1038/s41467-023-39655-3 ...up linked to whatever molecule you choose (Figure 1). Just express the fusion protein and add a specific...proteins, opening up new applications.   Figure 1: Live-cell labeling strategies. The green structure...minimally-disruptive genetic tag on the protein of interest. Table 1: Summary of popular self-labeling tag systems ...

...Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018...and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed Central...Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939-6371-3_3. PMID: 27730575; PMCID: PMC5270762. ... and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed PMID: 6337371. PubMed Central... system). The GAL80 repressor can bind to GAL4, partially inhibiting the binding of GAL4 to UAS. One can...domain (DBD) to GAL4 or VP16 activation domain results in chimeric GAL80-repressible and GAL80-insensitive...expression is low but is activated by GAL4 binding to UAS. If you place the GAL4 gene downstream of a tissue- ...

...authentic research experiences to undergraduate students (1, 2), but educators face significant challenges in ...6). Methionine synthesis in S. cerevisiae (Figure 1) occurs through a well-characterized pathway with ...sequences for ten different MET/Met genes (Figure 1) from the two yeast species into the pYES2.1/V5-His...selected from different parts of the Met pathway (Fig. 1). In addition to the complementation experiments that...about Clare at the Biology Dept. site.  References 1. American Association for the Advancement of Science...genes from the pYES2.1 plasmid is controlled by the GAL1 promoter. The cloning procedure adds both V5 and... to grow in the absence of methionine. Since the GAL1 promoter controls MET/Met gene expression, complementation...

...enzyme (among others): caveolin 1, also known as CAV 1. Usually, caveolin 1 promotes the cell life cycle ...caveolin 1, there’s less malignancy because there’s less proliferation. However, the lack of caveolin 1 in ...believes that miRNA may decrease the expression of TAK 1, an enzyme that controls apoptosis and regulates transcription...Agnieszka K., et al. "An absence of stromal caveolin-1 expression predicts early tumor recurrence and poor...museum "Fábrica Centro Ciência Viva" in Aveiro, Portugal. If cancer was a musician, then metastasize is...museum "Fábrica Centro Ciência Viva" in Aveiro, Portugal. She is particularly interested in science journalism...

...were found on the plasma membrane (top panel in Fig. 1). Of course, this was problematic, because the DREADD...plasma membranes in both species (bottom panel in Fig. 1). Our results indicate that the tag protein may have...protein of interest the membrane.   Figure 1: Electron microscope images showing ultrastructural...This post was contributed by Adriana Galvan, an associate professor at Emory University School of Medicine...express hM4Di in neurons of rhesus monkeys or mice (Galvan et al., 2019). The hM4Di receptor is an engineered....     Thank you to our guest blogger! Adriana Galvan is an associate professor at Emory University School...2159–2165 . https://doi.org/10.1128/mcb.8.5.2159 Galvan A, Raper J, Hu X, Paré J, Bonaventura J, Richie...

...for the wild-type. However, variants such as the P.1 (Gamma variant) and the B.1.351 (Beta variant) were... tested at the time of this study. Figure 1: Neutralization levels against spike pseudovirus decreases...lowest levels of cross-neutralization occurs in the P.1 and B.1.351 variants tested. Image from Garcia-Beltran...References Cele S, Gazy I, Jackson L, Hwa S-H, Tegally H, Lustig G, Giandhari J, Pillay S, Wilkinson E...Hanekom W, Moosa M-YS, Lessells RJ, de Oliveira T, Sigal A (2021) Escape of SARS-CoV-2 501Y.V2 from neutralization...Feldman J, Pavlovic MN, Gregory DJ, Poznansky MC, Sigal A, Schmidt AG, Iafrate AJ, Naranbhai V, Balazs AB...

...Weight Markers. Scientific Reports,7(1). doi:10.1038/s41598-017-02693-1. PubMed PMID: 28550309. PubMed Central...plasmids that can be used to make inexpensive 100 bp or 1 kb DNA molecular weight ladders were recently deposited... PstI or EcoRV, these plasmids generate 100 bp or 1 kb DNA ladders, respectively. Unlike many commercially... gels. Get Tips on Verifying Your Plasmid Table 1: Comparing Typical Comericial Ladders with the PSU...$10 to make 1000 lanes worth of the 100 bp or the 1 kb ladder. Using a commercial plasmid prep kit increases...Learn How to Analyze a Restriction Digest References 1. Henrici, R. C., Pecen, T. J., Johnston, J. L., & ...Additional Resources on the Addgene Blog Are you a Frugal Scientist? Save Money with the Baby Gel Box Find...

... you’d like to use to generate your new fly line: 1. Where would you like the gene to be incorporated ...control exactly in which locus your transgene ends up (1): Random insertion has the advantage that you can...he still reads “#” as a “pound symbol”. References 1. J. R. Bateman, et al. Site-Specific Transformation...experimental model organism. I then described the Gal4/UAS system used by geneticists to study gene function...favorite gene (YFG).  Sometimes, you want to use the Gal4/UAS system, but the available reagents do not match... or by filtering by fly as species of gene. Find Gal4 and UAS plasmids at Addgene Choosing a vector to...

...General Design Figure 1: Two proteins are fused to the FP(11) and FP(1-10) fragments. When Protein...and 11 to yield large (1-10) and small (11) fragments. In several cases an FP(1-10) fragment may be compatible...Description Article PI Blue EBFP2 EBFP2(1-10) and Capri(1-10) for use with GFP(11) Multiplexed labeling...Commun. 2017 Bo Huang mNeonGreen3 mNG3K(1-10) and mNG3A(1-10) for use with mNG2(11) Improved yellow-...sfCherry2 sfCherry2(1-10) and sfCherry2(11), also photo-activatable variant PAsfCherry2(1-10) Improved split...brings the pieces of the FP back together (Figure 1). This method is generally known as Bimolecular Fluorescence...even from a different base FP (for example, EBFP2(1-10) can be used with GFP(11)). Some of the articles...