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We narrowed to 337 results for: c rel

Showing: 81 - 100 of 337 results
  1. Fluorescent Biosensors for Measuring Autophagic Flux

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    ...comprised of two tandem fluorescent proteins: a relatively pH-insensitive RFP variant, DsRed.T3, that’s ...quenched in acidic environments, while mRFP is relatively stable and fluorescences even at the acidic pH... GFP/RFP ratio: a decrease in GFP fluorescence relative to RFP fluorescence suggests an increase in autophagic...autophagic flux, while an increase in GFP fluorescence relative to RFP fluorescence suggests a decrease in autophagic...∆G, however, can not be conjugated to PE due to a C-terminal glycine deletion, so it remains in the cytosol...changes in autophagic activity, and are a more reliable indicator of autophagic activity. The aim of this...important to remember that because Keima’s fluorescent relies on lysosomal acidity, this biosensor can’t be used...
  2. Hot Plasmids: Summer 2024

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    .... K., Chen, J., Pommier, G. C., Cogan, J. Z., Replogle, J. M., Adriaens, C., Ramadoss, G. N., Shi, Q.,...representative micrograph (C), particle orientation distribution and Fourier-shell correlation curve (D), and the... module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected with...Protein samples protected from AWI by helical LEAs. C-E) cryo-EM single-particle analysis of PRC2 with RvLEAMshort...tardigrades!  Find RvLEAMshort here!    Abe, K. M., & Lim, C. J. (2024). Small LEA proteins as an effective air-water...-02078-y.  Anzalone, A. V., Gao, X. D., Podracky, C. J., Nelson, A. T., Koblan, L. W., Raguram, A., Levy...Serack, F., Harvey, J. W., Brauer, P. P., Pirtle, C. P., Coffey, A., Howard, M., Kamath, N., … & Weissman...
  3. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    ...construct [1,2], i.e. positioning of the FP sequence relative to cell compartment targeting sequences, which...to characterize FP-fusion proteins functionally relative to the untagged protein of interest. To avoid ... retrieval sequence only functions at the extreme C-terminus [3]. You must therefore determine your cloning...strategy and decide whether you will create an N- or C-terminal fusion based on your experimental needs. ...
  4. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    ...changes from C•G-to-T•A are mediated by cytosine base editors (CBEs) and base changes from A•T-to-G•C are mediated...called point mutations that change C to T, G to A, A to G, and T to C, represent about two-thirds of known...Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464...Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR (2020...PACE and PANCE systems use M13 bacteriophage, which rely on the expression of gene III to create infectious...
  5. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

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    ...specific genomic locus: C-BERST Article contributed by Klaus Wanisch  Listen to the C-BERST podcast segment...processivity. Please find the MarathonRT plasmid here! Zhao C, et al. RNA. 2018 PubMed PMID:29109157. Nanobody ...Sontheimer lab used this technology, which they call C-BERST (dCas9–APEX2 biotinylation at genomic elements...in future studies of other genomic loci. Find the C-BERST plasmids here. Gao at al. Nature Methods (...protein fused with GFP, YFP, mCerulean, or other related fluorophores. Find all of the nanobody toolkit ...
  6. Important Considerations in Optogenetics Behavioral Experiments

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    ...response gene c-fos. For example, using Chr2 stimulation should result in robust increases in c-fos expression...stimulation compared to controls without Chr2. c-Fos is a reliable and widely used molecular marker of neuronal...proceed with your behavioral experiments. Comparison of c-fos amongst animals that receive different types of...neuronal activation, but it may still be possible to use c-fos expression to get the evidence you need. For example...activation in the novel context as determined by reduced c-fos expression, thus showing successful inhibition... as non-human primates [7, 8], zebrafish [9], and c. elegans [10]. If used properly, optogenetics is sure...literature to find all the applications of optogenetics relevant to your field. Ultimately, your question should...
  7. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

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    ...showed that iGluSnFR could detect neuronal activity in C. elegans, zebrafish, and mice. Second Generation ...Improved trafficking and localization through different C-terminal anchoring domains The original iGluSnFR in...adapted from Aggarwal et al… Figure 4. PDGFR is a C-terminal fusion to the PDGFR transmembrane domain ...mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol anchor. SGZ contains...domain, followed by a modified form of the cytosolic C-terminal domain of Stargazin including a terminal ...give researchers the ability to monitor glutamate release from neurons and other brain cells in vivo. Glutamate...differentiating between synaptic vs. extrasynaptic glutamate release. Building on the previous generation of the SF-...
  8. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ... standard strain, like DH5α, grown at 30 °C instead of 37 °C. If you’re unsure about the integrity of ...freshly harvested virus at 4 °C instead of storing it briefly at -80 °C. If troubleshooting, the safest...ultracentrifugation (75,000 - 225,000 g for 1.5 – 4 hours at 4 °C). After ultracentrifugation, virus may appear as a...freeze-thaw cycles on hepatitis B virus DNA and hepatitis C virus RNA quantification as measured with branched-DNA...restriction enzyme digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to ...reducing the culture volume into which the virus is released, a more concentrated virus solution can be obtained...
  9. Antibodies 101: Buffers, Storage, and Conjugates

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    ...can be very helpful if you're trying to find a relatively rare protein in an experiment with many proteins...short term storage (think days to weeks) in the 4°C. If you’ll only be using it every few month or if ...be going through small aliquots at a time, the -20°C is your best option, in aliquots no smaller than 10uL...
  10. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

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    ... present, it will repress expression of mKate2 relative to EBFP2. Weiss’s group used these reporters to...chain of BoNT/B into two fragments (BotNT-N and BotNT-C) and joined each fragment with one part of an established..., and showed that it can alter motor responses in C. elegans. Find the split BoTN/B plasmids at Addgene... it inactive, thus inhibiting neurotransmitter release. How is this light-mediated? The researchers split...visualizing the location and function of microbes rely heavily on fluorescent protein genes. These optical...
  11. Viral Vectors 101: Types of viruses

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    ...induce a lower inflammatory response within hosts. Relative to the other viruses discussed in this article...10.1016/j.gendis.2017.04.001 Maetzig T, Galla M, Baum C, Schambach A. Gammaretroviral vectors: biology, technology...2011 Jun;3(6):677-713. DOI: 10.3390/v3060677 Varanda C, Félix MDR, Campos MD, Materatski P. An Overview of...lentivirus and gamma-retrovirus are so closely related, their required components are very similar. While...
  12. 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...overview see Addgene’s AAV guide. AAV particles are relatively small (20 nm) in size which makes them capable... 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...
  13. Twenty Years of Sharing: Addgene's Viral Vector Service

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    ...could be a matter of timing, as jGCaMP7 was released relatively soon after GCaMP6. It could be a matter of...website.   Bibliography  Ersing, I., Rego, M., Wang, C., Zhang, Y., DeMaio, K. H., Tillgren, M., Fava, A.... E., Matho, K. S., Cetin, A., Woodard, K., Cepko, C., Guerin, K. I., Rego, M. A., Ersing, I., Bachle, ...10.3389/fnana.2019.00093 McDade, J. R., Waxmonsky, N. C., Swanson, L. E., & Fan, M. (2016). Practical Considerations...editor of Addgene’s eBooks — including the recently released Viral Vectors 101 2nd edition — I was incredibly...lessons learned from our plasmid lab to develop a reliable and robust high quality viral vector service, ...vector service because we realized that having reliable ready-to-use vectors made from plasmids in the...
  14. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    ...L-arabinose) without signs of toxicity or burden relative to a fluorescent protein control. We also added...visible in cells via TEM, and purified GVs under TEM. (c) Ultrasound imaging of tumor colonization by bARGSer-expressing...general method to create your own mARGAna cell line. (c) Z-projected phase contrast images of GV expressing...biomolecules. Appl. Phys. Lett. 110, 073704 (2017). Hurt, R. C. et al. Genomically mined acoustic reporter genes ...to produce GVs in their own lab using the most reliable method, which is the transient cotransfection ...
  15. Neurodegeneration Plasmid Collection

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    ...LRRK2 C-His LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40376 pBACgus/LRRK2 1867-2141 C-His...2161 C-His LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40378 pBACgus/LRRK2 1867-2176 C-His...2161 C-His_T2031E LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40381 pBACgus/LRRK2 C-His_G2019S...2161 C-His_G2019S LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40383 pBACgus/LRRK2 C-His_C2024S...mCherry-MAPTau-C-10 MAPT mCherry CMV Parkinson's, FTD Michael Davidson 55311 mTagBFP2-MAPTau-C-10 MAPT mTagBFP2... mKO2-MAPTau-C-10 MAPT mKO2 CMV Parkinson's, FTD Michael Davidson 58112 tdTomato-MAPTau-C-10 MAPT tdTomato...CMV ALS Alicia Pickrell 131792 pHAGE-TBK1-C-FLAG-HA TBK1 STOP codon CMV ALS Alicia Pickrell 131889 pDONR221...
  16. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

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    ...Gilmour, K. C., Adams, S., Thornhill, S. I., Parsley, K. L., Staal, F. J., Gale, R. E., Linch, D. C., Bayford...L., Quaye, M., Kinnon, C., Ancliff, P., Webb, D. K., Schmidt, M., von Kalle, C., Gaspar, H. B., Thrasher... PMCID: PMC7120651 Fu, Y., Foden, J. A., Khayter, C., Maeder, M. L., Reyon, D., Joung, J. K., & Sander... Mansour, M. R., Schwarzwaelder, K., Bartholomae, C., Hubank, M., Kempski, H., Brugman, M. H., Pike-Overzet...PMC189872 Schröder, A. R., Shinn, P., Chen, H., Berry, C., Ecker, J. R., & Bushman, F. (2002). HIV-1 integration...integration for their life cycle to continue. They rely on a viral protein called reverse transcriptase ...lentiviral vectors. This is because most titering methods rely on isolating genomic DNA from transduced cells and...
  17. Typing CRISPR Systems

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    ...Paez-Espino, D., Ma, E., Witte, I. P., Cofsky, J. C., Kyrpides, N. C., Banfield, J. F., & Doudna, J. A. (2018)... L. G., Wolf, Y. I., Covarrubias, P. C., Shmakov, S. A., Silva, C., Tolstoy, I., Johnson, D. B., Koonin...and poorly understood. There are three subtypes, A–C, and they are grouped together because they contain...component of CRISPR — a nuclease effector. However, IV-C does in fact have a helicase domain due to a gene ...CRISPR as a technology. There are three subtypes, A–C, with the beloved SpCas9 (from Streptococcus pyogenes...–842. https://doi.org/10.1126/science.aav4294 Hu, C., Myers, M. T., Zhou, X., Hou, Z., Lozen, M. L., Nam... activation and inactivation mechanisms in type I-C CRISPR-Cas3 for genome-editing applications. Molecular...
  18. Plasmids 101: Origin of Replication

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    ...pBR322 A Relaxed pColE1 ~15-20 ColE1 A Relaxed pR6K ~15-20 R6K* C Stringent pACYC ~10 p15A B Relaxed...pSC101 ~5 pSC101 C Stringent pBluescript ~300-500 ColE1 (derivative) and F1** A Relaxed pGEM ~300-500...held together with two hydrogen bonds not three as G-C pairs are. As a result, stretches of DNA that are ...number, ori, and incompatibility groups. Note the A -C compatibility grouping is an arbitrary designation...pMB1 (derivative) A Relaxed pBR322 ~15-20 pMB1 A Relaxed pET ~15-20 pBR322 A Relaxed pGEX ~15-20 pBR322...500 pUC and F1** A Relaxed pCDF ~20-40 CloDF13 (CDF) D Relaxed This table defines common cloning...replicate the host's chromosomal DNA, but they still rely on the host machinery to make additional copies....
  19. Fluorescent Tagging of Endogenous Genes with SapTrap

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    ...PCR or by evaluating visual phenotypes. Improving C. elegans fluorescent protein tagging with SapTrap ...called SapTrap to improve CRISPR genomic tagging in C. elegans. SapTrap offers scientists a convenient one...protein tagging systems For those who do not work with C. elegans, several groups have designed modular toolkits...by the Hornung Lab, allows users to easily create C-terminal tag fusions of endogenous genes in human ...plasmid containing the homology arms and an N- or C-terminal tag. The two PCR reactions are then mixed...line stably expressing Cas9. This PCR toolkit offers C- and N-terminal tagging vectors (eg GFP, Flag, YFP...the tag, selectable marker, and connector to be released (Fig. 1, Step 2-3). Since donor plasmids of the...
  20. Experimenting in an Open Source lab: from CRISPR to Cats

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    ...recent models of C. elegans molecular biology are based on the standard worm genome. While C. elegans are ...list of active spaces near you.   Learning about C. elegans with the OpenWorm Project One such space ...project. The main goal of OpenWorm is to simulate C. elegans as a computer model. When I first heard about...deconstruct and understand this complexity using existing C. elegans mutants. For instance, we use a mutant known...research goals in focus. Developing these skills relies upon biologists and computer scientists learning...neuron knockouts so that we can perturb factors related to worm motion. Working on the Cat Microbiome Project...
Showing: 81 - 100 of 337 results