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We narrowed to 282 results for: c rel

Showing: 101 - 120 of 282 results
  1. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...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...
  2. Multifunctional Neural Fibers: Modulation from Gut to Brain

    Type
    Blog Post
    Published
    Aug. 31, 2023, 1 p.m.
    ... the digestion of food, while brain tissue is relatively static). To operate both types of implanted ...it - the device has a rechargeable battery, is relatively small, and can be detached easily whenever necessary...Antonini, M.-J., Rosenfeld, D., Park, J., Garwood, I. C., Yan, W., Niroui, F., Fink, Y., Chandrakasan, A.,...monitor multiple systems. Ideally this tool would be wireless, able to monitor outcomes of interest, and stimulate...outputs for the system. Success! They also observed reliable optically evoked neuron activity for at least .... P. (2020). Modular Optoelectronic System for Wireless, Programmable Neuromodulation During Free Behavior... Multifunctional microelectronic fibers enable wireless modulation of gut and brain neural circuits. Nature...
  3. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...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... activation, and knockout. Most CRISPR screens rely on lentiviral systems, a fact supported by the wide...still beats lentivirus. In mice, AAV is almost entirely non-immunogenic — meaning when conducting in vivo...not integrate into the host genome. AAV instead relies mainly on episomal expression. Episomes are circular...It is important to note, however, that while AAV rarely integrates into the genome, CRISPR may increase...
  4. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C., & Telesnitsky, A. (2008). Pseudodiploid Genome...34372494. Prussin, A. J., Garcia, E. B., & Marr, L. C. (2015). Total Virus and Bacteria Concentrations in...Xiao, H., Conway, J. A., Hunter, E., & Kappes, J. C. (1997). Functional RT and IN incorporated into HIV...tools in molecular biology. “Retroviruses” is an umbrella term which encompasses 1.) the gamma-retroviruses...After uncoating, reverse transcription follows the release of the two strands of viral RNA and of the viral...The genome of viral tools are often not virology related at all, and you will instead get expression of ...genome can direct the production, packaging, and release of progeny viral particles. And there you have ...
  5. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    Published
    Nov. 12, 2020, 2:15 p.m.
    ...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...usage important? Translation and protein synthesis relies on transfer RNAs (tRNAs). tRNAs bind to and deliver...studies have shown that translational efficiency is correlated with codon bias across all endogenous genes in...you want to express may contain codons that are rarely used in the host you are using or come from an ...
  6. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    Published
    Nov. 5, 2024, 8:32 p.m.
    .... Time Viral production in mammalian cells is relatively quick. After transfection, HEK-293T cells start...https://doi.org/10.1096/fj.01-0995fje. Tan, E., Chin, C. S. H., Lim, Z. F. S., & Ng, S. K. (2021). HEK293 ...doi.org/10.3389/fbioe.2021.796991. Urabe, M., Ding, C., & Kotin, R.M. (2002). Insect cells as a factory ... cells, and its lifecycle ends in cell lysis, releasing the virus to the supernatant. The virus will reach...
  7. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ... use next-generation sequencing to measure the relative proportion of guide RNAs (gRNAs) across the population...our example later, histones) ChiRP-seq RNA Hi-C DNA (genome-to-genome interactions) DNase-seq DNA...commonly think of DNA as a linear polymer of A,T,G, and C nucleotides arranged in a double helix. People are...die or differentiate Step 3 reveals which NANOG-related genes are critical to stem cell survival. Knowing...
  8. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...and purification An affinity tag, generally a relatively small sequence of amino acids, is basically a...tag to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses of the affinity ... tags, a His tag can be fused to either the N- or C-terminus of a protein. Unlike other epitope tags –... Thus far Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes used ...crystallizing it or testing its function. Small ubiquitin-related modifier (SUMO) can help with folding and stabilization...western blots, IP, co-IP, IF, flow -cytometry, but rarely used for purification as elution requires low pH...  0.8 Affinity and Purification Very small size, rarely affects function S-tag KETAAAKFERQHMDS  1.8 Solubility...
  9. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...both methods. These approaches have relatively low sensitivity, which can be problematic if you are trying...also influence antibody binding by altering the relative concentration of potential antibody binders and...little to no signal in the liver sample, which you do (C), so your antibody is validated! Genetic Strategies...10.1080/19420862.2015.1100787  Taussig MJ, Fonseca C, Trimmer JS (2018) Antibody validation: A view from...recommendations for improving antibody use and reliability (Roncador, et al., 2015; Taussig, et al., 2018...your target then you would expect to see good correlation between the levels of target detected in both... targeting a different epitope, and look for correlation between the results. Using antibodies against...
  10. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...expressed to identify successfully transfected cells. C-to-T editing by the APOBEC-Cas9 editosome would restore...codon that is converted to a ‘TAC’ or ‘TAT’ after a C-to-T base editing event. This edit changes the histidine... ATG codon from an ACG codon using a base-editor (C>T conversion) they could induce the translation of...Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464...re inefficient compared to CRISPR/Cas9 edits that rely on double strand DNA breaks. Due to this inefficiency...2018). TLDR: The ACE reporter from the Harris Lab relies on the restoration of a frame-shift in mCherry ...quantifies APOBEC-Cas9 efficiency in real-time, it relies on DSBs which is not ideal for monitoring single...
  11. Easing the Protein Purification Process with pCri

    Type
    Blog Post
    Published
    June 19, 2015, 3:08 p.m.
    ...requires a substantial amount of properly folded, relatively pure protein, but getting to this stage is often...plasmids with different varieties of tags, both N- and C-terminal. Each set of plasmids uses the same restriction...
  12. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    Published
    June 1, 2023, 1:15 p.m.
    ... Sellnow, R. C., Boye, S. L., Coberly, B., Bennett, A., Agbandje-McKenna, M., Sortwell, C. E., Hauswirth...Backer, M. W. A., Brans, M. A. D., Luijendijk, M. C., Garner, K. M., & Adan, R. A. H. (2010). Optimization...Dudek, A. M., Pillay, S., Puschnik, A. S., Nagamine, C. M., Cheng, F., Qiu, J., Carette, J. E., & Vandenberghe..., J. Y., Han, J. H., Lee, H., Han, Y. E., Rah, J. C., & Park, H. (2020). Labeling dual presynaptic inputs... Porrot, F., Bohl, D., Marmé, F., Chérel, Y., Salvetti, A., Hurtrel, B., Heard, J.-M., Rivière, Y., & ...your specific experimental system, rather than relying on other systems as proxies.  Finally, the size...consider. First, the titer required is inversely related to transduction efficacy: the higher the efficiency...
  13. Antibodies 101: Epitope Tags

    Type
    Blog Post
    Published
    May 11, 2023, 1:15 p.m.
    ...Epitope tags are short peptides introduced at the N or C terminus of a protein that are bound by antibodies...enterokinase, which recognizes the 5 amino acids on the C terminal of the tag. Of the most common tags, FLAG...not at all. The introduction of a tag at the N or C terminus of a protein may disrupt the biological function...a specific application, or they exist but are unreliable for a variety of reasons (Baker, Nature). Tagging...
  14. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    Published
    Sept. 3, 2019, 1:15 p.m.
    ...please sign up here.        New toolkit for N or C terminal tagging of proteins for degradation using...has created a new toolkit of plasmids for the N- or C- terminal tagging of proteins with mAID and other ...cytidine deaminase  Target-AID-NG, which mediated a C to T conversion in the target sites adjacent to an...Nureki’s lab engineered SpCas9 (SpCas9-NG) with relaxed preferences for the third base of the PAM so that...
  15. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...Zhi Ying Chen, Robert C. Robbins, Mark A. Kay, Michael T. Longaker, and Joseph C. Wu. "A nonviral minicircle... generating iPSCs. The generation of iPSCs is relatively simple in concept: ectopically express a cocktail.... The advantages of this method are that it’s relatively simple to implement and doesn’t require time-...a Sendai Virus-Based Vector Using a Nonstructural C Protein and Endogenous MicroRNAs." Plos One 11, no...Overall, integrating viral vectors efficiently and reliably generate iPSCs, but they have several safety concerns...important control. Additionally, Sendai virus doesn’t rely on promoters for transcriptional regulation, so ...Check Out Our Interview with Stem Cell Researcher Darrell Kotton Four Factors that Differentiate the Stem...
  16. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    Published
    May 23, 2023, 1:15 p.m.
    ... Boeckman, J. X., Manning, C. F., Nguyen, J. T., Bechtold, H., Dumitras, C., Gong, B., Nguyen, K., Van...-GABA-AR β1 (white) and anti-GABA-AR β3 (green). (C) Hippocampus labeled with anti-Kv2.1 (magenta), anti-AnkyrinG... Panels C1-C3 show magnified detail of the box in C. Image adapted from Andrews et al., 2019 under CC-... J., Kandov, E., Lowekamp, B., Speranza, E., Chu, C. J., Gola, A., Thakur, N., Shih, R., Yao, L., Yaniv... colocalization factor to assess the spatial relationship between the targets. Or, it might be sufficient...
  17. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...Oh S, Tomkinson AE, Hendrickson EA, Giovannangeli C, Jasin M, Brunet E (2014) Chromosomal Translocations.../10.1016/j.molcel.2014.08.002 Ran FA, Hsu PD, Lin C-Y, Gootenberg JS, Konermann S, Trevino AE, Scott DA... Yan M-Y, Li S-S, Ding X-Y, Guo X-P, Jin Q, Sun Y-C (2020) A CRISPR-Assisted Nonhomologous End-Joining... and how these pathways work, as this could be relevant when planning your experiment. Here, we’ll discuss...repair is that the Cas9 protein doesn’t immediately release from the broken end after cleavage, which may interfere...
  18. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    Published
    Feb. 24, 2015, 7:50 p.m.
    ...to fuse the activators instead of at the protein’s C terminus. With that new design in place, Konermann...Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki...Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann ... not managed to create a system to allow us to reliably activate essentially any gene,” she says. The ...tools because obviously, unlike the mouse, we can’t rely on genetic engineering to optimize humans for CRISPRi...
  19. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...should the double strand break (DSB) be positioned relative to where the homology ends? A9: The Zhang lab ...10.1126/science.1231143  Elliott, B., Richardson, C., Winderbaum, J., Nickoloff, J. A., & Jasin, M. (1998....1128/mcb.18.1.93  Ran, F. A., Hsu, P. D., Lin, C., Gootenberg, J. S., Konermann, S., Trevino, A. E....j.cell.2011.06.019  Wang, H., Yang, H., Shivalila, C. S., Dawlaty, M. M., Cheng, A. W., Zhang, F., & Jaenisch...j.cell.2013.04.025  Yang, H., Wang, H., Shivalila, C. S., Cheng, A. W., Shi, L., & Jaenisch, R. (2013)....
Showing: 101 - 120 of 282 results