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We narrowed to 321 results for: POR C

Showing: 101 - 150 of 321 results

  1. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    Published
    March 28, 2024, 1:15 p.m.
    ...conformations: (A) bent-closed, (B) extended-closed, and (C) extended-open. When a ligand binds, it typically ... with much lower affinity than their open states (C). Figure courtesy of IPI. IPI anti-integrin toolset...and resources References Hao, Y., Yan, J., Fraser, C., Jiang, A., Anuganti, M., Zhang, R., Lloyd, K., Jardine...Jaumouillé, V., Hao, Y., Coppola, J., Yan, J., Waterman, C. M., Springer, T. A., & Ha, T. (2022). Single-molecule...in competition with other similar integrins, as reported by IPI and the Springer lab (Hao et al., 2024)...In this study, we, along with the Springer lab, reported the binding specificity, kinetics, and affinity...belief that αVβ3 would be the most functionally important integrin on these cells. What’s next IPI is working...

  2. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    .../000504294 Wang, T., Guan, C., Guo, J., Liu, B., Wu, Y., Xie, Z., Zhang, C., & Xing, X.-H. (2018). Pooled...doi.org/10.13918/j.issn.2095-8137.2016.4.237 McCusker, C., & Gardiner, D. M. (2011). The Axolotl Model for ...elife.50651 Vieira, W. A., Wells, K. M., & McCusker, C. D. (2019). Advancements to the Axolotl Model for ... areas of microbial research like metabolism. Transport, biofilms, and much more. Because of it’s BSL-...designed for use in Arabidopsis. There are also important tips for plant transformations and the use of ...

  3. Click Biology: How to Click and Play to Explore Biology

    Type
    Blog Post
    Published
    Oct. 21, 2025, 1:15 p.m.
    ...Cysteine-less, fast split intein; N-terminal CLm and C-terminal Aes gp41-1 Split inteins A small, fast ...attB2r-SNAP-attB3 SNAP-tag Gateway entry clone to add C-terminal SNAP-tag Promega Plasmid Collection HaloTag...PCV-Cas9, Cas9-PCV HUH-tag Cas9 plasmids with N- or C-terminal PCV2 HUH-tag; increase homology-directed ... M. R. (2025). Click biology highlights the opportunities from reliable biological reactions. Nature Chemical...

  4. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...store them in a -80°C freezer to prevent RNA degradation. They will degrade in a -20°C freezer.   RNA extraction...DEPC. Alternatively, glassware can be baked at 180°C for at least 4 hours.   If your final RNA sample(s...extractions- if you need really clean RNA, it’s important to make sure that when extracting, to only take...

  5. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ...cloning method with high throughput capability. Engler C, Kandzia R, Marillonnet S. PLoS One 2008;3(11):e3647..., Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic...2011 Apr 14. PubMed.   Golden Gate cloning. Engler C, Marillonnet S. Methods Mol Biol. 2014;1116:119-31...like Gateway cloning or Gibson assembly. Another important consideration is the design of flanking overhangs...

  6. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ...fragments: N-terminal BirA* (N-BirA*) and C-terminal BiraA* (C-BirA*). These two fragments are fused to...also which fragment of BirA*, either the N- or the C-terminus, the bait proteins were tagged with. It’s...interacting proteins. No spatiotemporal control. Many PPI detection methods lack spatiotemporal control or an “... lysate fractionation. Important considerations for Split-BioID It’s important to remember that with Split-BioID...this limitation by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent...The Split-BioID method Enter Split-BioID, a spatiotemporally controllable version of the BioID protocol... other PPI methods, including BioID, is its spatiotemporal controlled labeling of interacting proteins...

  8. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ...length of your assay. The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most...to incubate their primary antibody overnight at 4 °C and their secondary antibody for 1–2 hrs at room temperature...the DIY method. As some proteins aggregate at 95 °C, you may want to check for any available data on the...expensive, reduce protocol time, can be stored at 4 °C, and are very consistent gel-to-gel.  Ladder Whatever...stored at your bench during the procedure and at 4 °C when you’re not running a western. They are not very...will noticeably degrade after several months at 4 °C.  Pro tip! If you get a speckled image, your protein-based...variety of pore sizes. If your protein of interest is small, you may want to consider a smaller pore size (...

  9. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...Williams, Eric J. Ross, Sofia M. C. Robb, Sarah A. Elliott, Kimberly C. Tu, Alejandro Sánchez Alvarado,...discovered and characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific...Hybridization (WISH) WISH is a technique that detects the temporal and spatial localization of mRNA transcripts in...

  10. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...still unexpectedly affect nearby genes (Figure 1A-C). Find CRISPR plasmids at Addgene! Knock-in: Introducing...site (TSS) deletion. (B) Mutations of splice sites. (C) Removal of an exon or a large genomic fragment. (... doi:10.1080/15476286.2014.996067 Basak J, Nithin C. Targeting Non-Coding RNAs in Plants with the CRISPR-Cas...experiments are important for visualizing ncRNA localization and function, but directly incorporating very long...transcribed but do not encode proteins. They play important roles in many cellular and regulatory processes...complex and not yet fully understood, so it is important to carefully select the target locations of knock-in...nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription machinery of a target...

  11. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...   References and Resources   References Beck C, Zhang D, Gong Y. Enhanced genetically encoded voltage...: 10.1016/j.cobme.2019.10.010. Bando, Y., Grimm, C., Cornejo, V.H. et al. Genetic voltage indicators.... 71 (2019). doi:10.1186/s12915-019-0682-0. Beck, C., Gong, Y. A high-speed, bright, red fluorescent voltage...sensors are better classified by their fluorescent reporter and the mechanism of their voltage sensing component...rhodopsin proteins were initially evolved for ion transport, not fluorescence, they are not nearly as bright...for penetrating deeper tissues depths (1 mm), important for certain applications such as in vivo tissue...rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET.  Image from ...

  12. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...doi.org/10.1038/mt.2009.280 Hirsch ML, Li C, Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski....2010.205 Trapani I, Colella P, Sommella A, Iodice C, Cesi G, Simone S, Marrocco E, Rossi S, Giunti M, ...Trapani I, Toriello E, de Simone S, Colella P, Iodice C, Polishchuk EV, Sommella A, Colecchi L, Rossi S, Simonelli...1964 . https://doi.org/10.1172/jci34316 Bak RO, Porteus MH (2017) CRISPR-Mediated Integration of Large ...Large Gene Cassettes Using AAV Donor Vectors. Cell Reports 20:750–756 . https://doi.org/10.1016/j.celrep.2017.06.064...

  13. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    Published
    July 13, 2017, 2:30 p.m.
    ... 25°C. At low temperatures like 18°C, flies take longer to develop. At high temperatures like 30°C, flies...combs. To make genetic crosses properly, it is important to isolate females from males while they are still... after hatching into their adult phase. It is important to clear out the parent crosses from the container...feature when setting up genetic crosses. More importantly, this knowledge will stop you from embarking ...

  14. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...Adenine base editors (ABE) mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make...Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464...Lee S-J, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G (2020...selection of guideRNAs. Of particular note, ABE8s reported in this work did not cause genome-wide, guide-...guide-independent off-target deamination, an important attribute for therapeutic application. ABE8s demonstrate...

  15. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...four reprogramming factors- OCT4, SOX2, KLF4, and C-MYC - that induce a differentiated somatic cell to...additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is one of the reprogramming ...22645348. PubMed Central PMCID: PMC3390875. 25. Patsch, C., et al., Generation of vascular endothelial and smooth... PubMed Central PMCID: PMC4363833. 35. Eguizabal, C., et al., Dedifferentiation, transdifferentiation,...1): p. 82-94. PubMed PMID: 23329641. 36. Jopling, C., S. Boue, and J.C. Izpisua Belmonte, Dedifferentiation... adenovirus [5], Sendai virus [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]...insulin-producing β-like-cells [37, 38]. In 1987, Davis et al. reported one of the earliest examples of transdifferentiation...

  16. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ... should already be at 37 °C. Most labs have a water bath constantly at 37°C for this purpose.Learn about... Cells usually have their comfort zone around 37 °C and 5% CO2 and get agitated with sudden changes in...passage are too young and should not be used for important transfection experiments right after they’ve been...

  17. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ... P., Rodriguez-Aguayo, C., Walton, B. L., Sood, A. K., Greaver, J., Janssen, C. F., & Lopez-Berestein,... Academic Press. Turner, P. V., Brabb, T., Pekow, C., & Vasbinder, M. A. (2011). Administration of substances...muscle groups (usually the thigh), though it is important to avoid injection into the sciatic nerve. Mice...

  18. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...assembly of NgAgo/ssDNA requires incubation at 55 °C - a dangerous, non-physiological temperature for mammalian...genome or just after a stop codon (as required for C-terminus epitope tagging). NgAgo survey Since it was..., he tries random things (101 lessons on Spanish, C++, guitar, photography) and often dreams of a sunny...Researchers from a variety of institutions recently reported their inability to recapitulate the results of...

  19. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...GFP (sfGFP) and mNeonGFP can fold in <10min at 37°C, mCherry takes ~15min, TagRFP ~100min and DsRed ~10hours...instance, enhanced GFP (EGFP) was optimized for 37°C, and is therefore most suited for mammalian or bacteria...GFPS65T is better suited for yeast studies (24-30°C). Brightness: Brightness is a measure of how bright...temperature). pH Stability: This parameter is important if you are planning to express the FP in acidic...

  20. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...TurboID and APEX2 at different cellular locations. C) Enrichment of dual-labeled proteins identifies proteins...bioRxiv (A) and Qin and Cheah et al. 2023 bioRxiv (B-C). They were able to label proteins moving to and...fiber labeled with HaloTag-iPAK4. Scale bar 10 µm. C) Images of iPAK4 fibers in cultured rat hippocampal... editing efficiencies in vitro and in vivo. Incorporating reverse transcriptase (RT) variants, PE6a and...., 2023, reused under CC-BY 4.0 License. Importantly, even commercial antibodies can be tethered on... more. For added precision, the authors also incorporated LOV-Turbo into TransitID experiments.  Find ...in light-regulated LOV-Turbo enables precise spatiotemporal control of proximity labeling in living cells...

  21. IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging

    Type
    Blog Post
    Published
    June 8, 2023, 1:15 p.m.
    ...References and Resources References Radtke, A. J., Chu, C. J., Yaniv, Z., Yao, L., Marr, J., Beuschel, R. T.... J., Kandov, E., Lowekamp, B., Speranza, E., Chu, C. J., Gola, A., Thakur, N., Shih, R., Yao, L., Yaniv...along with the ability for any user to upload supporting documentation that agrees or disagrees with the...

  22. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...site for a total of 2 binding sites per antibody. C) IgG’s are big enough that multiple labels (green ... IgG secondary antibody (Pleiner et al figures 4A-C). And since they have limited cross-reactivity between...Bates, M., Böhning, M.I., Chug, H., Görlich, D., Lee, C., Pleiner, T., Schliep, J.E., Stark, H., Trakhanov... genetic engineering of a tag or fusion with a reporter enzyme. For multicolor staining, careful consideration...al. Before picking a secondary nanobody, it’s important to review a few characteristics of your primary...

  23. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ... Cas9 into n-terminal (Cas9(N)-2xNLS) and c-terminal (Cas9(C)-2xNLS) fragments facilitates target DNA ...obtain even more precise temporal control of gene knockout or activation, the c-terminal cas9 fragment ... the nuclear export sequence. Treatment with rapamycin induces Cas9(N)-FRB-NES and Cas9(C)-FKBP-2xNLS ... (dCas9(C)-FKBP-2xNLS-VP64 and dCas9(N)-FRB-NES). This system provides users greater temporal control ...an n-terminal DNA recognition domain and a mostly c-terminal nuclease domain. Feng Zhang's group utilized...fragment was fused with FK 506 binding protein 12 (Cas9(C)-FKBP-2xNLS) and the n-terminal cas9 fragment with ...constructing fusion proteins with customizable N- and C-terminal modules   Thanks to the lab of Tomás Santalucía...

  24. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    Published
    Aug. 5, 2025, 1:15 p.m.
    ...McMahon, C., Baier, A. S., Pascolutti, R., Wegrecki, M., Zheng, S., Ong, J. X., Erlandson, S. C., Hilger...    References and Resources References Chan, A. C., & Carter, P. J. (2010). Therapeutic antibodies for..., S. G. F., Ring, A. M., Manglik, A., & Kruse, A. C. (2018). Yeast surface display platform for rapid ...means improving research. More specific antibodies support common research tools like western blots, ELISAs...

  25. New Norepinephrine Indicators: nLightG and nLightR

    Type
    Blog Post
    Published
    Dec. 12, 2023, 2:15 p.m.
    ...References and resources References Feng, J., Zhang, C., Lischinsky, J. E., Jing, M., Zhou, J., Wang, H.,...j.neuron.2019.02.037 Kagiampaki, Z., Rohner, V., Kiss, C., Curreli, S., Dieter, A., Wilhelm, M., Harada, M....Neuromodulators like dopamine and norepinephrine have important functions in the brain but have been difficult... are blocked by alpha1-AR antagonist. This is important because alpha2-AR can provide inhibitory feedback...

  26. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...) Strategy for leak-free transcriptional control. C) Iterative transduction workflow overview. Image from...nucleases identified, functions at temperatures up to 43 °C and can be used for genome editing in moderate thermophiles...straightforward workflow for doing gene targeting, reporter generation, controllable gene knockouts, and inducible... models, toolkits like this one provide new opportunities for investigating genetic mechanisms related...

  27. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ...constructed 22 different mTFP1 fusions to both the C- and N- termini of the fluorescent protein and found... PubMed Central PMCID: PMC2292683. Shaner, Nathan C., et al. "A bright monomeric green fluorescent protein...enhanced GFP (EGFP)-type termini. mNeonGreen was reported as the brightest monomeric green or yellow fluorescent... or for live cell imaging of fusions and FRET reporters. mTFP1 is also insensitive to physiologically ...

  28. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ...the annealing temperature of the PCR from 60° C to 61° C improved assay reliability. Different primers... PubMed Central PMCID: PMC2957240. 5.Taylor, Sean C., Genevieve Laperriere, and Hugo Germain. "Droplet... people usually report? Addgene and other viral vector manufacturing facilities report the physical titer... characteristics give us information on three important factors. What can AAV titers tell you? Physical...cell transduction assays. Wild-type AAV2 has been reported to have a near-perfect physical-to-infectious ... However, for recombinant AAV2, the same study reported a physical-to-infectious particle ratio of 50:...(“full”) (Zeltner et al., 2010), while another reported that only 20% of recombinant AAV2 particles were...

  29. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ...from this publication at Addgene. 2. Swarts, Daan C., et al.  “DNA-guided DNA interference by a prokaryotic...from this publication at Addgene. 3. Swarts, Daan C., et al. “Argonaute of the archaeon Pyrococcus furiosus...NgAgo In eukaryotes, Argonaute proteins play an important role in RNA interference, binding RNA guides to...

  30. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    Published
    July 30, 2024, 1:15 p.m.
    ...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...from 2016–2023 by type of tool. Equally as important to what is being delivered is where it’s being...vectors not only available, but accessible. The supporting data and community engagement required to do ...lessons learned from this process have shaped and supported our growth and development. We’ve shared what ...

  31. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...10.1126/science.1206593 Powers S, Miranda C, Dennys-Rivers C, Huffenberger A, Braun L, Rinaldi F, Wein....2020.107878 Sinnett SE, Hector RD, Gadalla KKE, Heindel C, Chen D, Zaric V, Bailey MES, Cobb SR, Gray SJ (2017... More recently, the Bird lab found that only a portion of the MECP2 protein is necessary to rescue the...Neurological Disorder by Programmable RNA Editing. Cell Reports 32:107878 . https://doi.org/10.1016/j.celrep.2020.107878...

  32. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...editing window, for example all C’s will edit to T’s. Thus, it’s important that there are no “bystander ... single base pair edit. (Cytosine deaminases edit C-to-T and adenosine deaminases edit A-to-G.) In short...takes place on the free strand of DNA and converts a C to U without producing a double strand break. ... particular gene isn’t feasible. For example, temporary repression or knockdown allow you to study essential...

  33. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    Published
    May 6, 2014, 1:07 p.m.
    ... by isolating a calcium-binding protein, troponin C (TnC), from muscle cells. They identified a toadfish...calcium indicators can be used in mouse, Drosophila, C. elegans, and zebrafish – essentially all of the standard...said. “We know CREB is really important, but where is it really important and when? That is not known.”...calcium-binding sites as possible for added sensitivity. As reported in Nature Methods, the FRET changes of these Twitch...

  34. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    Published
    July 29, 2025, 1:15 p.m.
    ...Maher, M. F., Nasti, R. A., Vollbrecht, M., Starker, C. G., Clark, M. D., & Voytas, D. F. (2019). Plant gene...0337-2  Molla, K. A., Sretenovic, S., Bansal, K. C., & Qi, Y. (2021). Precise plant genome editing using...Gao, Q., Liu, S., Zhang, X., Hu, W., Wang, Y., Han, C., Li, D., & Wang, X. (2025). Transgene- and tissue...these reagents in both plant and animal systems. Importantly, they allow transient delivery, enabling transgene-free...

  35. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ...nitrogen or a dry ice/ethanol bath, and store at -80° C. Once thawed, the AAV aliquot should be used quickly...PubMed Central PMCID: PMC190186. 9. Grieger, Joshua C., and Richard J. Samulski. "Packaging capacity of ...Axonal Transport It has been demonstrated that certain serotypes can undergo axoplasmic transport. These...of neurons (retrograde transport) or towards the synapse (anterograde transport), which is a possible ...retrograde and anterograde transport, but they also have similar transport characteristics (velocities...strong CNS tropism (4). If axonal transport is not desirable, transport can be reduced by injecting a smaller...and using AAVs have in common? The three most important items when looking for a house and choosing an...

  36. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...mEosFP’s chromophore requires a temperature below 30°C and this is not ideal for experiments in mammalian...can form it’s chromophore at a more comfortable 37°C (5). Dendra2 photoconverts from green to red fluorescence... pH stability, chromophore maturation rate at 37 °C, turnover rate, and the level of background fluorescence...: 1035-1040. PubMed PMID:18724362. 8.Stiel, Andre C., et al. "Generation of monomeric reversibly switchable...impossible to determine specific protein turnover or temporal expression using standard FP-chimeric proteins...second. The first successful irreversible PA-FP reported was PA-GFP. PA-GFP was derived from Jellyfish ...localization, or leading to protein aggregation. Another important parameter to consider is the contrast ratio- a...

  37. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    Published
    April 16, 2020, 1 p.m.
    ...Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY (2020) ...Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung... https://doi.org/10.1126/science.aaq0179 Myhrvold C, Freije CA, Gootenberg JS, Abudayyeh OO, Metsky HC... to cleave and activate a reporter. The detectable signal from this reporter can be measured and quantified...and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...variants, Cas13, and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect ...method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2) (Gootenberg...

  38. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    Published
    Jan. 4, 2022, 2:15 p.m.
    ...create solution C. And if a conflict resolver does their job well, not only will solution C be better than...many sources of conflict are substantive (“Person C told Person A one thing and inadvertently told Person... is customer support, but there are eight other teams who still retain a customer support element amongst... your technical skills, can open new career opportunities for you, if you can identify them in yourself...explaining my approach to conflict resolution, it’s important to recognize that there are different types of...or goal. Resolving these types of conflicts is important for the individuals involved, and also for the...responsibilities. Knowing which aspects of customer support each team “owns” is critical for day-to-day operations...

  39. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    Published
    May 28, 2020, 1:15 p.m.
    ...constructs 69570 and 69571 together encode the N and C terminal segments respectively of a GFP dependent ...-N-CretrcintG Connie Cepko 69571 AAV1 pAAV-EF1a-C-CreintG Connie Cepko 75469 AAV1 pAAV-EF1a-Flp-DOG-NW.... The advantages of optogenetics lie in the spatiotemporal control offered by the system that allows for... expression of ChR2(H134R) bound to an mCherry reporter. We now also offer AAVrg for plasmid 83898, which...advantage of the extensive supply of transgenic reporter lines that supply a variety of GFP expression ...

  40. A Control for All Seasons

    Type
    Blog Post
    Published
    Oct. 5, 2023, 1:15 p.m.
    ...replicates such as multiple mice in each group and C, technical replicates or repeated measurements will...interactions between the sample and the secondary antibody. C, a label control treats the sample to the full protocol...PubMed Central PMCID: PMC5773976. Waters, Jennifer C. “Accuracy and Precision in Quantitative Fluorescence...binding  Primary antibody specificity is further supported by a pre-immune serum or isotype-specific control...

  41. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    Published
    June 15, 2023, 1:15 p.m.
    ...come out! References Noren, C. J., Anthony-Cahill, S. J., Griffith, M. C., & Schultz, P. G. (1989). A...describing a technique that allowed for intentional incorporation of an unnatural amino acid in translation (Noren...natural translation machinery; these are often incorporated into the cells via plasmid. Third, a second ...containing the chosen codon that will result in the incorporation of the non-canonical amino acid at the site ...integrated into the cell’s genome, instead of incorporation via plasmid or media. Theoretically, if you ...will produce a protein with a site specifically incorporated ncAA, ready for you do to any number of experiments... They provide resources, knowledge, and most importantly, validated protocols (using readily available...

  42. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    Published
    Aug. 4, 2020, 1:15 p.m.
    ...Pewe L, Wohlford-Lenane C, Hickey M, Manzel L, Shi L, Netland J, Jia HP, Halabi C, Sigmund CD, Meyerholz...SARS-CoV-2 were developed for broad use and are now important for accelerating research into this virus. One...during the pandemic and in the future, depends on support from the scientific community. Deposits of new ... be deposited into repositories, and financial support to maintain repositories’ capacity for storing,...all ways in which the scientific community can support repositories. Repositories are essential for the...

  43. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ... a mutation that causes Niemann-Pick disease type C, directly demonstrating the therapeutic potential ...toxicity in long-term experiments.  Figure 4: The C-terminus of Cre (CreC) and the N-terminus of Cre (...rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET. Image from Beck...indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed PMID: 31685893. A MoClo toolkit for... Eisenhoffer lab, is a new tool that increases temporal and spatial precision of gene expression and reduces...

  44. Exploring Applications of the Bioluminescent HiBiT Tag

    Type
    Blog Post
    Published
    Oct. 2, 2025, 1:15 p.m.
    ...Machleidt, T., Robers, M. B., Benink, H. A., Eggers, C. T., Slater, M. R., Meisenheimer, P. L., Klaubert,...657–661. https://doi.org/10.1038/nmeth.4344  Peach, C. J., Kilpatrick, L. E., Woolard, J., & Hill, S. J....-proteins-using-hibit-crispr-cell-lines/ Shibata, C., Otsuka, M., Shimizu, T., Seimiya, T., Kishikawa,...Figure 1). Addition of substrate generates a proportionate luminescent signal, strong enough to detect ...signal to gradually diminish over time; and, importantly, repeated excitation generating reactive oxygen...targets in translational research Researchers can incorporate HiBiT with a wide range of workflows, from classic...paper for more detail on whether the authors incorporated HiBiT by CRISPR tagging at the endogenous locus...

  45. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...disorder of glycosylation, and Niemann-Pick disease type C. Although the three DMD studies referenced here represent... 2015 Dec 31. doi:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H, Sanchez-Ortiz...K, Cheng JK, Chew WL, Widrick JJ, Yan WX, Maesner C, Wu EY, Xiao R, Ran FA, Cong L, Zhang F, Vandenberghe...optimized, it’s important to ascertain how long the rescue phenotype will last, and more importantly, if it does...genetic disease in a postnatal mouse model, an important proof of concept for future preclinical and clinical...does not cause obvious germline modification, an important finding given the controversy associated with ...great step forward for CRISPR gene therapy, it’s important to realize that DMD is a simpler case than other...

  46. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...also shown that the Q system is also functional in C. elegans. Subsequent work by the Luo and Potter labs...23995928; PMCID: PMC3811519.Seo SO, Schmidt-Dannert C. Development of a synthetic cumate-inducible gene ...; PMCID: PMC5270762. Wendler, F., Park, S., Hill, C. et al. A LexAop > UAS > QUAS trimeric plasmid to ...stage-specific promoter and design a UAS reporter construct, the reporter will only be expressed where and/or...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...the Q system from a gene cluster in the fungus Neurospora crassa. The QUAS promoter is less leaky than ... transcription from the UAS promoter. This is important because it allows them to be used together for...

  47. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    Published
    Feb. 1, 2022, 2:15 p.m.
    ...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...whenever you use the antibody. This information is important, and you’ll need to track it throughout your experiments...

  48. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    Published
    June 6, 2023, 1:15 p.m.
    ...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...Lenti particles enter the nucleus through nuclear pores and thus do not require breakdown of the nuclear...

  49. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ... of cytidine and adenine base editors that enable C-to-T and A-to-G base substitutions respectively. Substitutions...:723–728 . https://doi.org/10.1021/sb500351f Dong C, Fontana J, Patel A, Carothers JM, Zalatan JG (2018... https://doi.org/10.1016/j.cell.2013.02.022 Ronda C, Pedersen LE, Sommer MOA, Nielsen AT (2016) CRMAGE...Vridhambal GS, Rivera AJ, Montgomery SA, Fortier L-C, Barrangou R, Theriot CM, Ousterout DG (2020) In Vivo...the phage recombineering machinery are first electroporated with pCas9. Then, pCRISPR is introduced along... a mammalian cell line (Qi et al., 2013). Two important features of this early system are its regulatable...Euler CW, Jiang W, Nussenzweig PM, Goldberg GW, Duportet X, Fischetti VA, Marraffini LA (2014) Exploiting...

  50. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...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)....refer to Cong et al. (2013) for more details. One important note is that this ‘split RNA’ design is not as...). One-Step generation of mice carrying reporter and conditional alleles by CRISPR/CAS-Mediated Genome...
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