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We narrowed to 337 results for: c rel

Showing: 61 - 80 of 337 results
  1. Hot Plasmids and Viral Preps - July 2021

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    Blog Post
    ...activity on NAG PAMS and reduced activity on NGG PAMs relative to wild type. A new genome-wide CRISPR knockout...) 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...opportunities for investigating genetic mechanisms related to morphogenesis, cell fate specification, and ...the internal control (Fig 1). Because this method relies on the fluorescence ratio between the 2 fluorophores...
  2. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ... amplification in the host cell. This enables relatively high and long-term expression of the reprogramming...the reprogramming factors- OCT3/4, SOX2, KLF4, and C-MYC [1]. For the derivation of clinical grade hiPSCs...vectors that encoded six factors - OCT3/4, SOX2, KLF4, C-MYC, LIN28 and SV40 large T antigen (SV40LT). The ...of TP53 enhances iPSC generation as does replacing C-MYC with L-MYC [10, 11]. While the mechanisms underlying...understood, the addition of an shRNA for P53 and replacing C-MYC with L-MYC in the episomal reprogramming cocktail...patients are reprogrammed less efficiently, no such correlation has been found between the age of the patients...Models for Disease & Open Science: Interview with Darrell Kotton Four Factors that Differentiate the Stem...
  3. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

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    Blog Post
    ...fragments: N-terminal BirA* (N-BirA*) and C-terminal BiraA* (C-BirA*). These two fragments are fused to... are enriched relative to controls using Split-BioID-GFP and BioID run on six unrelated proteins. One ...also which fragment of BirA*, either the N- or the C-terminus, the bait proteins were tagged with. It’s...These interactions are often modeled as binary relationships, i.e. protein A interacts with protein B; but...transient PPIs. Many PPI identification techniques rely on protein interactors to stay in close contact ...
  4. Design Tips for Prime Editing

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    Blog Post
    ... with 40–60% G/C content are most likely to be successful, although sequences with G/C content outside...of the 3′ extension of the pegRNA should not be C. A C base is speculated to base pair with G81 of the..., J., Oyler-Castrillo, P., Ravisankar, P., Ward, C. C., Levesque, S., Jing, Y., Simpson, D., Zhao, A.,...Figure 3: pegRNAs whose 3′ extension begins with a C base can cause unwanted base pairing (left), while... Koblan, L. W., Levy, J. M., Chen, P. J., Wilson, C., Newby, G. A., Raguram, A., & Liu, D. R. (2019). ... Knipping, F., Ravisankar, P., Chen, P.-F., Chen, C., Nelson, J. W., Newby, G. A., Sahin, M., Osborn, ..., Anzalone, A. V., An, M., Newby, G. A., Chen, J. C., Hsu, A., & Liu, D. R. (2022). Engineered pegRNAs...
  5. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...Time: ~40 minutes Equipment required: 37°C heat block and 62°C heat block Approach: Viral RNA is extracted...range Time: ~1 hour Equipment required: 37°C water bath and 42°C water bath Approach: RNA is extracted from...10.1038/s41596-019-0210-2 Lucia, C., Federico, P.-B., & Alejandra, G. C. (2020). An ultrasensitive, rapid...Valois, E., Solley, S. C., Braig, F., Lach, R. S., Baxter, N. J., Kosik, K. S., Arias, C., Acosta-Alvear, D...incubation  Time: ~90 minutes Equipment needed: 37°C heat block, LED blue light illuminator or UV light... in a range of temperatures varying between 10-50°C. Visit Addgene's COVID-19 Plasmids and Resources ...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) ...
  6. Rinehart Lab Phosphoprotein Reagents

    Type
    Collection
    ...Ter Haar, C. M., Rogulina, S., Amrofell, M. B., Isaacs, F. J., Rinehart, J., & Jewett, M. C. (2015). Robust...mCherry, while a phosphobinding domain is fused to the C-terminal mCherry fragment. Upon interaction between...Link opens in a new window) Barber, K. W., Miller, C. J., Jun, J. W., Lou, H. J., Turk, B. E., & Rinehart..., T., Guo, L., Osborne, E. M., Benner, J., Noren, C. J., Rinehart, J., & Söll, D. (2011). Expanding the...synthesis via partial UAG codon reassignment and release factor 1 deletion . FEBS Letters , 586 (20), 3716...
  7. Protocol - Bacterial Transformation

    Type
    Protocol
    ...Shaking incubator at 37 °C Stationary incubator at 37 °C Water bath at 42 °C Ice bucket filled with ice... storage at 4°C and let warm up to room temperature and then (optional) incubate in 37°C incubator. Mix...also make their own competent cells. This is a relatively simple procedure and is useful for performing...transform Procedure Take competent cells out of -80°C and thaw on ice (approximately 20-30 mins). Remove...placing the bottom 1/2 to 2/3 of the tube into a 42°C water bath for 30-60 secs (45 secs is usually ideal...without antibiotic) to the bacteria and grow in 37°C shaking incubator for 45 min. Pro-Tip This outgrowth...and won't grow in colonies. Incubate plates at 37°C overnight. Tips and FAQ How can I save time when carrying...
  8. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ... 25°C. At low temperatures like 18°C, flies take longer to develop. At high temperatures like 30°C, flies...genomes Compared to humans and mice, flies have a relatively simple genome. They have four chromosomes: the...females do not. Also, males have sex combs on their forelegs which are black, bristly hairs. Females do not...
  9. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    Blog Post
    ...While this method can be time-consuming, it is relatively effective in reducing off-target effects since...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...
  10. An Introduction to Adenovirus

    Type
    Blog Post
    ...., Pan, C. Y., Rizzo, K., Bertumen, J. B., Morales, C., Oluniyi, P. E., Nguyen, J., … Chiu, C. Y. (2023...36996871. Tatsis, N., Fitzgerald, J. C., Reyes-Sandoval, A., Harris-McCoy, K. C., Hensley, S. E., Zhou, D., Lin..., Iparraguirre, A., Lopez-Camacho, C., Wherry, E. J., & Ertl, H. C. J. (2007). Adenoviral vectors persist...Borkenhagen, L. K., Fieldhouse, J. K., Seto, D., & Gray, G. C. (2019). Are adenoviruses zoonotic? A systematic review....org/10.1016/B978-0-12-801238-3.02523-X. Hofmann, C., Löser, P., Cichon, G., Arnold, W., Both, G. W., ...AdVs are ubiquitous (Schmitz et al., 1983), yet rarely cause severe disease in humans (and no vaccines...stages of infection for efficient cell lysis and release of adenovirus from infected cells. J Virol, 70(...
  11. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...2018). For C>T and A>G base editors, the intended edit must be in a 5 - 10 nt window relative to the PAM...target sites that code for amino acids close to the C’ terminus of the protein, to maximize the chances ...bystander edits are possible if there is another target C or A in the window. Another technology, prime editing...doi.org/10.1038/s41587-020-0561-9 Dempster JM, Pacini C, Pantel S, Behan FM, Green T, Krill-Burger J, Beaver...JL, De Los Angeles A, Mali P, Aach J, Kim-Kiselak C, Briggs AW, Rios X, Huang P-Y, Daley G, Church G (...introduction of a specific mutation, one generally relies on homology directed repair (HDR) to incorporate...along with the single cell clone (although this is rarely done). When targeting a dsDNA break for HDR, the...
  12. Lentivirus ddPCR Titration

    Type
    Protocol
    ...parameters: Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Enzyme Activation 95 10...titer: $$T = {V*C*D\over v}$$ Where: T = Infectious titer, TU/mL V = Viruses per genome C = # Cells per ... across the dilutions. RPP30 copies should be relatively constant across samples. In the RRE example below...concentration of RPP30 positive droplets stays relatively even across samples (green). To increase the ...Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 Pierceable...Benzonase in DMEM complete to each well. Incubate at 37 °C for 30 min. Gently aspirate media from wells, using...can be used for ddPCR immediately or stored at -20 °C until ready to use. Preparing for ddPCR Thaw samples...
  13. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

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    Blog Post
    ...raw %}{"no_follow":false,"open_in_new_tab":true,"rel":"noopener","sponsored":false,"url":{"content_id"...https://doi.org/10.1016/j.cell.2025.05.002 Hunker, A. C., Wirthlin, M. E., Gill, G., Johansen, N. J., Hooper...Larsen, R., Gore, B. B., Szelenyi, E. R., Trader, C., . . . Tasic, B. (2021). Enhancer viruses for combinatorial...Somasundaram, S., Miller, J. A., Kalmbach, B. E., Radaelli, C., Gore, B. B., Weed, N., Omstead, V., Bishaw, Y., ...10.1007/s00018-021-03903-w  Thomas, H. F., & Buecker, C. (2023). What is an enhancer? BioEssays, 45(10). https...knowledge. We need to continue to develop improved, reliable tools to help carry out that research. Organizations...
  14. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...jcm.23.5.959-961.1986 Lobstein, J., Emrich, C. A., Jeans, C., Faulkner, M., Riggs, P., & Berkmen, M. (2012... some people still use amp? Carb is expensive (relative to amp), and for some applications, the benefit...References Hwang, J. M., Piccinini, T. E., Lammel, C. J., Hadley, W. K., & Brooks, G. F. (1986). Effect...their mode of action and have a very similar relationship as amp and carb (they are interchangeable). ...low rate, there is evidence that zeocin is never entirely detoxified, even when Sh ble is expressed (Trastoy...
  15. Hot Plasmids: Fall 2024

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    Blog Post
    ...Optimized mScarlet-based tags for live imaging in C. elegans CRISPR/Cas9 for gene disruption in M. abscessus...tagging proteins, as it can be fused to the N- or C-terminus (or even in the middle of proteins) and performs.... E., Young, D. F., Goswami, K. K., & Russell, W. C. (1987). Isolation and characterization of monoclonal...39406745. Optimizing mScarlet tags for live imaging in C. elegans By Alyssa Neuhaus Red fluorescent proteins...maturation rate limits its use in systems like the C. elegans embryo, where developmental processes happen...recently generated new constructs specifically for C. elegans (Cao et al., 2024). The team designed coding...mScarlet3 and mScarlet-I3 that were codon-optimized for C. elegans and included artificial introns to improve...
  16. Cancer and the Immune System: Deciphering the Relationship

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    Blog Post
    ...T-regs) into their microenvironment by producing C-C motif chemokine 22 (CCL22) and TGFβ. T-regs are suppressive...tolerance-producing antigens enunciated above. Finally, signals released from the microenvironment help create tolerance...eradicate metastasis or eliminate the probability of a relapse. Chemical and radiation therapies cause a transient...
  17. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...CyPet with C-terminal His tag SCFP3A Cyan Mammalian Express a gene of interest fused to the C-terminus...YPet with C-terminal His tag SYFP2 Yellow Mammalian Express a gene of interest fused to the C-terminus...Mammalian Express a gene of interest fused to the C-terminus of monomeric Cerulean mTurquoise2 Cyan ...Mammalian Express a gene of interest fused to the C-terminus of Amber mVenus N1 Yellow Mammalian Express...Mammalian Express a gene of interest fused to the C-terminus of monomeric Venus pCEP4YPet-MAMM Yellow...Mammalian Express a gene of interest fused to the C-terminus of LSSmOrange mRuby2 Red Mammalian Expresses...FRET occurs. By photobleaching the acceptor, you release the donor's quenching and the fluorescence of the...
  18. Predicting Adverse Reactions to Monoclonal Antibody Drugs

    Type
    Blog Post
    ...Bryson, C., Fogg, M., Baker, M. P., Herskovitz, J., Goletz, T. J., Zhou, L., Moxness, M., Flynn, G. C., Narhi...trained on vast amounts of data to calculate the relative immunogenicity score (Mattei et.al., 2022).    ...Antidrug Antibody Formation in Oncology: Clinical Relevance and Challenges. The oncologist, 21(10), 1260–1268...
  19. Reflections and Looking Ahead

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    Blog Post
    ... D. W., Kumaran, R., Rego, M., Schofield, C., … Laflamme, C. (2024). A consensus platform for antibody... H., Laflamme, C., Houston, D. W., Polinski, N. K., Meijers, R., Levey, A. I., Saper, C. B., Errington...Ayoubi, R., Ryan, J., Gonzalez Bolivar, S., Alende, C., Ruiz Moleon, V., Fotouhi, M., Alqazzaz, M., Southern... strides in making science more accessible and reliable. We continue to expand our educational content...
  20. Viral Vectors 101: Producing Your rAAV

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    Blog Post
    ...S., Croft, C. L., Futch, H. S., Ryu, D., Ceballos-Diaz, C., Liu, X., Paterno, G., Mejia, C., Deng, D.,...References  Aurnhammer, C., Haase, M., Muether, N., Hausl, M., Rauschhuber, C., Huber, I., Nitschko, H...Esbin, M. N., Whitney, O., Walther, N., & Maurer, A. C. (2023). Transgene Expression in Cultured Cells Using...., Chtarto, A., Clark, K. R., Conlon, T., Darmon, C., Doria, M., Douar, A., Flotte, T. R., Francis, J....serotype, as some rAAV serotypes are abundantly released into the culture media, while others remain cell-associated...Wilson, J. M. (2010). Efficient serotype-dependent release of functional vector into the culture medium during...
Showing: 61 - 80 of 337 results