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We narrowed to 400 results for: POR C

Showing: 81 - 100 of 400 results
  1. To Each HIS Own

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    Blog Post
    ...histidine amino acids on the N- or C- terminus of a protein. Neither the N or C terminus is preferable over ...histidine residues expressed in frame on the N- or C-terminus of a recombinant protein of interest. The...need to decide whether to add the tag to the N- or C-terminus and how many histidine residues to include...tended to be very large and often required the incorporation of a cleavage site and a post-purification cleavage...very pure sample. Because binding strength is proportional to the length of the tag, with longer tags you...
  2. Viral Vectors 101: Optogenetic Tools

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    Blog Post
    ..., Kadmon, J., Raja, C., Chibukhchyan, A., Ramakrishnan, C., Inoue, M., Shane, J. C., McKnight, D. J., ...Mattis, J., Tye, K. M., Ferenczi, E. A., Ramakrishnan, C., O’Shea, D. J., Prakash, R., Gunaydin, L. A., Hyun...A., Pieper, F., Sims, R., Tenedini, F., Bergs, A. C. F., Bendifallah, I., Zhou, F., Zeitzschel, N., Ahlbeck...individual types of GPCRs with greater spatial and temporal control than is achieved by traditional pharmacologic...photoswitchable, orthogonal, remotely-tethered ligands (PORTLs). OptoXRs are chimeras of the extracellular domain...change shape with light exposure to activate them. PORTLs use a similar concept, except the ligand is attached...
  3. CRISPR History and Development for Genome Engineering

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    ...and others) Drosophila Plants (monocots and dicots) C. elegans Yeast ( S. cerevisiae and S. pombe ) Zebrafish...Zebrafish Xenopus References Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero ...23287718 Dalvai M, Loehr J, Jacquet K, Huard CC, Roques C, Herst P, Côté J, Doyon Y. 2015. A Scalable Genome-Editing-Based...Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann ...libraries can be used to identify genes that are important to a given phenotype. Current libraries are available...
  4. A History of Genome Engineering in Popular Culture

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    Blog Post
    ...rise of literary giants such as Isaac Asimov, Arthur C. Clarke, James Blish, and John W. Campbell. This period...combination of the four bases of DNA – A, T, G, and C. In this movie Ethan Hawke’s character revolts against...editors (CBE) use a cytidine deaminase to introduce a C→T change at precise locations in the genome. This ...JR, Sousa AA, Koblan LW, Levy JM, Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR (2019) Search-and-replace...Correction: Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 559:E8...420–424 . https://doi.org/10.1038/nature17946 Lenay C (2000) Hugo De Vries: from the theory of intracellular...editing technology led to the development of many important medical and agricultural products. But it also...
  5. Plan Your Experiment

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    ...Whitehead, E. H., Guimaraes, C., Panning, B., Ploegh, H. L., Bassik, M. C., Qi, L. S., Kampmann, M., &...Delivery Validate Genetic Edit References CRISPR ( C lustered R egularly I nterspaced S hort P alindromic...25408407 Dixit, A., Parnas, O., Li, B., Chen, J., Fulco, C. P., Jerby-Arnon, L., Marjanovic, N. D., Dionne, D...testing multiple pegRNA designs is particularly important for prime editing. CRISPR Intereference : Target...Predicted On-Target and Off-Target Activity Another important factor to keep in mind when designing your gRNA...In addition to off-target activity, it is also important to consider factors that maximize cleavage of ...generally perform by chemical transfection or electroporation. Expression can be transient, or you can generate...
  6. Hot Plasmids - October 2020

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    Blog Post
    ... synaptic proteins Fluorescent protein system for C. elegans Fluorescent timers for studying cell cycle...native locus, but introducing large genetic changes in C. elegans with CRISPR can be difficult since it requires...can you use CRISPR to introduce large FP genes into C. elegans? By using a split FP approach. Split-wrmScarlet...together, they fuse and fluoresce. The team engineered C. elegans cell lines to express wrmScarlet1-10 unattached...grown in high concentrations of IL-3 show a high proportion of blue fluorescence, indicating cell proliferation...they could not proliferate. Eastman et al. Cell Reports 2020. https://doi.org/10.1016/j.celrep.2020.107804...
  7. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

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    ...02708-5 Hrit J, Goodrich L, Li C, et al. (2018) OGT binds a conserved C-terminal domain of TET1 to regulate...89–92. https://doi.org/10.1038/301089a0. Galonska C, Charlton J, Mattei AL et al (2018). Genome-wide tracking...https://doi.org/10.7554/eLife.34870.  Kaas GA, Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR...sigh. The problem is that many of these studies report correlations without clearly testing whether a ...Sapozhnikov and Szyf 2021, Charlton et al. 2020). Most importantly, TET proteins affect transcription independent...
  8. Plasmids 101: Biotinylation

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    Blog Post
    ...PMC3223083. 6. McInnes, J. L., Forster, A. C., Skingle, D. C., & Symons, R. H. “Preparation and uses of...PMC164612. 11. Liou, Y.-R., Wang, Y.-H., Lee, C.-Y., & Li, P.-C. “Buoyancy-Activated Cell Sorting Using Targeted...fusion in which AviTag is added to either the N or C terminus of the protein. BirA has been shown to effectively...of carboxyl groups to organic acids to produce important cellular metabolites. Biotin acts as a carboxyl...primary antibody respectively. Avidin bound to a reporter is then added to the reaction for easy detection... experiments. During detection, a biotinylated reporter enzyme that is pre-incubated with free avidin ...protein of interest leading to amplification of the reporter signal and easier detection. This strategy is ...
  9. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

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    ...labeling of the cell population. Figure 2: Dense (c), intermediate (d), and sparse (e) labeling of the...References: del Valle Rodríguez A, Didiano D, Desplan C (2011) Power tools for gene expression and clonal ...Feng Q, Zhou Y, Zeng S, Ren M, Jiang S, Ni H, Zhou C, Gong H, Luo M (2018) Cell-type-specific and projection-specific...was developed by Thomas Clandinin’s lab, and incorporates several common genetic tools in Drosophila (...: a method to genetically manipulate precise proportions of cells. bioRxiv 788679; https://doi.org/10.1101...
  10. p53 Pathway

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    ...RFWD2); E3 ubiquitin protein ligase cytoC Cytochrome c DR5 Tumor necrosis factor receptor superfamily, member...Ghebranious N, Igelmann H, Lu X, Soron G, Cooper B, Brayton C, Park SH, Thompson T, Karsenty G, Bradley A, Donehower... of p53’s pro-apoptotic effects is especially important to tumorigenesis. Inheriting only one functional...
  11. Optogenetics + CRISPR, Using Light to Control Genome Editing

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    Blog Post
    ...fragment of CIB1 and it was fused to both the N- and C-termini of dCas9; and 2) CRY2FL-VP64, a fusion of ...fragmented Cas9 into N-terminal (residues 2-713, N713) and C-terminal (residues 714-1368, C714) halves, rendering...split-Cas9 (FAST) system constitutively expresses the C-terminal fragment of Cas9 while transcription of the...used the bacterial phytochrome BphS which produces c-di-GMP in response to far-red light (730 nm). This...VP64 which in turn transcribes N-Cas9. The N- and C-Cas9 fragments then dimerize to form a functional ... from A. sativa (Bubeck et al., 2018). LOV2 has a C-terminal helix that unfolds when stimulated by blue...Bietz A, Waldhauer MC, Börner K, Fakhiri J, Schmelas C, Dietz L, Grimm D, Correia BE, Eils R, Niopek D (2018...
  12. 27 Hot Plasmids from 2016

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    ... 79628 (C) Cv (capsid) N and C 79633 (N) and 79641 (C) NS1 (var: W98G) N and C 79635 NS3 C 79637...79637 NS2B C 79636 NS4A N 79640 NS4B C 79639 NS5 C 79632 (N) and 79631 (C) PrM N and C   Detecting... for N-terminal fusions and Myc tagged BioID2 for C-terminal fusions. Kim, et al. Mol Biol Cell. 2016...streamline CRISPR/Cas9-based genetic tag insertion in the C. elegans genome, Erik Jorgensen’s lab from the University...paramagnetic ferritin protein were tethered to the C-terminus of TRPV4 (a pressure-sensitive channel). ...Rodríguez-Laureano L, Albanese SK, Gradia S, Jeans C, Seeliger MA, Levinson NM, Chodera JD. bioRxiv preprint...et al. Cell. 2015. PubMed PMID: 26627737 pORTMAGE: A portable method for bacterial genome engineering ...
  13. The Pleiades Promoter Project

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    ...., Chen, Y., Cheng, J. C., Chopra, V., Docking, T. R., Dreolini, L., D'Souza, C. A., Flynn, E. K., Glenn...intron-lacZ/NLS References Portales-Casamar, E., Swanson, D. J., Liu, L., de Leeuw, C. N., Banks, K. G., Ho...plasmid webpages. These plasmids are described in Portales-Casamar et al., 2010 . List of Pleiades MiniPromoters...MiniPromoters MiniPromoter Source Gene Construct Reporter Ple2 ADORA2A pEMS1142 EGFP/NLS Ple3 ADORA2A pEMS1143...
  14. Plasmids 101: Multicistronic Vectors

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    ... L L T C G D V E E N P G P P2A: (GSG) A T N F S L L K Q A G D V E E N P G P E2A: (GSG) Q C T N Y A...ribosome skip the synthesis of a peptide bond at the C-terminus of a 2A element, leading to separation between...between the Glycine and Proline residues found on the C-terminus meaning the upstream cistron will have a ...along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to ...sometimes advantageous, particularly when only a portion of the plasmid is packaged for viral delivery, ...expression levels between two or more genes is important. How do multicistronic vectors work? Scientists...single transcript. The two strategies most widely incorporated into plasmids for research purposes are described...
  15. Simplify Cloning with in vivo Assembly

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    Blog Post
    ... added to the PCR mix to remove parental DNA (37 °C for 15 mins). This enzyme specifically cuts methylated...region, with a melting temperature (Tm) of around 60 °C for all primers. The length of the homologous region...recombination. We normally use a Tm between 47-55 °C for the homologous region (calculated using Oligo ...product. For example, by combining multiple primers, a C-terminal epitope tag can be moved to the N-terminus...pathway is ubiquitous, with successful recombination reported in all laboratory E. coli strains tested so far...use of this pathway as a cloning tool was first reported over 25 years ago (Douglas and Howard, 1991, Bubeck...cloning strain has failed so far. What is most important is the transformation efficiency of the strain...
  16. To Codon Optimize or Not: That is the Question

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    Blog Post
    ...cells. For example, Henry Lester’s lab optimized the C. elegans GluCl ion channel genes for expression in...Santana-Quintero LV, Katneni U, Simonyan V, Kimchi-Sarfaty C (2017) A new and updated resource for codon usage ...–34 . https://doi.org/10.1038/nsmb1031 Gustafsson C, Govindarajan S, Minshull J (2004) Codon bias and ...doi.org/10.1093/nar/28.1.292 Sauna ZE, Kimchi-Sarfaty C (2011) Understanding the contribution of synonymous... in a particular organism. Why is codon usage important? Translation and protein synthesis relies on transfer...deliver amino acids to the ribosome where they are incorporated into the growing polypeptide chain. One part...even been linked to some diseases. It has been reported that 5-10% of human genes contain a region where...
  17. Using AAV for Neuronal Tracing

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    ...J.G., Wohnoutka, P., Gerfen, C.R., Koch, C., Bernard, A., Dang, C., Jones, A.R., and Zeng, H. (2014). A ..., J. M., Vandenberghe, L. H., Haute, C. V., Thiry, I., Deroose, C. M., Mortelmans, L., Wilson, J.M., Debyser... Ravi, N., Wu, W.-L., Sánchez-Guardado, L., Lois, C., Mazmanian, S.K., Deverman, B.E., and Gradinaru, ...Winslow, B., Cain, N., Mihalas, S., Wang, Q., Lau, C., Kuan, L., Henry, A.M., Mortrud, M.T., Ouellette,... E., Tanaka, M., Kearney, M., Chattree, G., Yang, C. F., Shah, N.M., and Mooney, R. (2017). Identification...Res 406(1-2), 317-321. PubMed PMID: 2436717. Towne, C., Schneider, B.L., Kieran, D., Redmond, D.E.Jr., and... facilitate transport: retrograde transport relies on dynein, while anterograde transport relies on kinesin...
  18. The AAVantages of AAV in CRISPR Screens

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    ...References and Resources References Braun, C. J., Adames, A. C., Saur, D., & Rad, R. (2022). Tutorial: design...Sousa, A. A., Tsai, S. Q., Bengtsson, N. E., Lööv, C., Ingelsson, M., Chamberlain, J. S., Corey, D. P.,...M. J., Joung, J. K., Breakefield, X. O., Maguire, C. A., & György, B. (2019). High levels of AAV vector...CRISPR screens rely on lentiviral systems, a fact supported by the wide array of lentiviral-based CRISPR libraries...locations that could cause deleterious effects. It is important to note, however, that while AAV rarely integrates..., CRISPR may increase this rate. Specifically, portions of the AAV genome can integrate at double-stranded...
  19. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

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    ... Koblan, L. W., Levy, J. M., Chen, P. J., Wilson, C., Newby, G. A., Raguram, A., & Liu, D. R. (2019). ..., X., Chen, W., Martin, B. K., Calderon, D., Lee, C., Choi, J., Chardon, F. M., McDiarmid, T. A., Daza...Kissling, L., Marquart, K. F., Schmidheini, L., Solari, C., Balázs, Z., Krauthammer, M., & Schwank, G. (2023...-024-02268-2 Schep, R., Brinkman, E. K., Leemans, C., Vergara, X., van der Weide, R. H., Morris, B., van... In our study, we used the TRIP (Thousands of Reporters Integrated in Parallel; Akhtar et al., 2013; Schep...Chromatin Position Effects Assayed by Thousands of Reporters Integrated in Parallel. Cell, 154(4), 914–927....
  20. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

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    Blog Post
    ...resistance, b) the promoter that drives the insert, and c) any other selection markers (for use in other cell...pBACKBONE-Flag-hGene If there was also an EGFP fused to the C-terminal of your insert you would list it after the...identify what your plasmid contains and other important information. Tip: A lowercase "p" is often used...simple piece of information can often convey many important details. Once you know the backbone a plasmid ...
Showing: 81 - 100 of 400 results