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We narrowed to 466 results for: cat.3

Showing: 181 - 200 of 466 results

  1. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    Published
    Feb. 28, 2017, 3:30 p.m.
    ...PMC4364232. Find the plasmids from this publication at Addgene. 3. Ma, Hanhui, et al. “Multiplexed labeling...produce yellow, cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created... every genomic locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot induce gene editing...Cas9 ortholog, and the target sequence must be located adjacent to that PAM. To scale up fluorescent CRISPR...this rainbow of colors, Ma et al. performed sophisticated tracking of multiple chromosomal loci in live... do not induce genome editing. If used with catalytically active Cas9, the short, CRISPRainbow gRNA will... PMCID: PMC3918502. Find plasmids from this publication at Addgene. 2. Ma, Hanhui, et al. “Multicolor...

  2. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ...shown below, a fragment with 5’ overhang TGGA and 3’ overhang TCCG can be ligated into a vector containing...to specify assembly order, so fragments with 5’ or 3’ sequence homology cannot be assembled using this ... work around this is to "domesticate" your fragment: PCR-based amplification can be used to create silent...and exciting applications, this system is an incredibly powerful tool for cloning complicated constructs...Type IIS sites (such as BsaI or BbsI), that are located on the outside of the cleavage site. As a result...plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning...restriction sites that may not be amenable to "domestication", you might want to consider using an alternative...

  3. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...genetically targetable fluorescent indicators. Nature protocols. 2006 Aug;1(3):1057-65. Resources on Addgene.org...(2) allosteric rearrangements of the sensor, and (3) subsequent change in fluorescence only occurs at ... a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium...in living cells and organisms. Unlike chemical indicators, these DNA constructs can be delivered to intact...the sensors discussed below fall into this GECI category.   For calcium sensors, one size doesn...:509-18. Inoue M. Genetically encoded calcium indicators to probe complex brain circuit dynamics in vivo...Kanemaru K, Iino M. Genetically encoded fluorescent indicators for organellar calcium imaging. Biophysical journal...

  4. Celebrating 15 Years of Scientific Sharing

    Type
    Blog Post
    Published
    Jan. 14, 2019, 2:08 p.m.
    ...like The Atlantic. We’ve been in documentaries in 3 languages (Japanese, German, and English) and even...we need to do it quickly. Hiring is hard but as a 3 year Boston Globe and BBJ Best Place to Work with ...enough space! There’s enough space here at our new location to grow. We are hiring again and getting ready...to keep up with the demand with excellence and dedication. We don’t have worry or wonder if providing...research, hosting conferences, and are expanding our catalog of materials to better serve the community. We ...projects allows us to support more new projects. Diversification is happening. The Addgene team will have some...we might do. I am grateful every day for their dedication, flexibility, innovative thinking, and openness...

  5. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    Published
    July 25, 2017, 1:20 p.m.
    ...pulse of 500 nm light (turn on psRaf1) followed by 3 seconds of 400 nm light (turn off psRaf1). After this...is responsible for this MEK1/2 negative feedback. 3. Studying signaling pathways in vivo Expressing photoswitchable...members of their pathways, but where the kinases are located, their signal strength, and how long their signals...and MEK1 is active or “on.” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when...domains were inserted at sites homologous to the locations they were attached to in MEK. The photoswitchable...kinases good for? There are a number of potential applications for photoswitchable kinases.  Below are a few...signaling by using psMEK1 and an ERK kinase translocation reporter with a mRuby2 fluorescent reporter ...

  6. Human Germline Editing Using CRISPR

    Type
    Blog Post
    Published
    Aug. 10, 2017, 2:19 p.m.
    ...place before DNA replication. Indeed, they observed that 41/42 embryos were non-mosaic at 3 days post fertilization...showed a 3% mosaic rate, and this rate would need to be much, much lower before clinical applications could... embryos carried indels at the mutation site, indicating that repair proceeded through NHEJ rather than...attributable to CRISPR editing. Conclusions & implications One striking feature of this paper is the failure...experiments are needed before we can consider clinical applications for germline editing. Although this paper represents...

  7. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    Published
    Jan. 31, 2019, 1:32 p.m.
    ...microenvironments made of?  A tumor’s genesis is divided into 3 phases: initiation, progression, and metastasis. The...microenvironment’s physiology is deeply connected to these 3 stages and contains multiple cell types: fibroblasts... these stromal cell modifications, stromal cells are also constantly communicating with a tumor, proving... was contributed by Bárbara Pinho, a science communicator at the Portuguese science museum "Fábrica Centro...Proficient in transportation, exosomes are tools of communication and provide malignancy to the microenvironment... Pinho! Bárbara Pinho is currently a science communicator at the Portuguese science museum "Fábrica Centro...

  8. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...an improved design of the guide RNA that caps the 3’ end with a structured RNA motif. These engineered...editing tools, the desired edits are templated by the 3′ extension of the pegRNAs. PEDAR is capable of correcting...AsCas12f1. They found that this system, without any modifications, is effective in genome editing in both bacteria...therapeutically relevant gene targets and expand the application to the possible treatment of genetic disease....

  9. New Videos: Addgene Lab Tips

    Type
    Blog Post
    Published
    Feb. 15, 2018, 2:41 p.m.
    ...help ensure ensure equal distribution. Lab tip #3: Barcode everything Use barcodes and scanners to keep...With barcode tracking, you will always be able to locate materials quickly and easily. Learn more about ...Additional Resources on the Addgene Blog New Educational Resource: The Addgene Videos Page 5 Science Rap...

  10. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...Minimal Tol2 or miniTol2 is a truncated version of the original Tol2 and has ~3-fold increase in transposition...the QCascade DNA-targeting module with the crRNA, 3) a transposase complex made up by three transposase... up to 10 kB in size in bacteria.   Figure 3: INTEGRATE system for RNA guided transposon insertion... It generally stays put, which makes it easy to locate in a genome. But there is a type of mobile DNA,... hyperactive and likes to jump around from one location in the genome to another. This jumping is what...most transposable elements (TEs) results in the duplication of this target site sequence at the site of insertion...This is similar to how retroviruses, like HIV, replicate. Class I TEs do not encode a transposase enzyme...

  11. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    Published
    Aug. 23, 2022, 1:15 p.m.
    ... motivated reasoning. Psychological bulletin, 108(3), 480. More resources on the Addgene blog Summer SciComm...Further, when science is communicated in a format that is accessible, that communication often neglects the ...SciCommSummer Scicomm Series: Modes of CommunicationCareers in Science Communication: Science Writing... to save a few days’ worth of capsules to self-medicate with when they get a cold. A primary school teacher...dihydrogen monoxide (H2O). Studying science communication as a PhD student Fortunately, by the time I ...Otago had established the Centre for Science communication and I was accepted as a grad student. I chose...a phenomenon called “motivated reasoning,” the catchall term for a range of neat little tricks our minds...

  12. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...GpNLuc and OgNLuc are available through Addgene (Cat.# 70185 and 70186, respectively). Antares Chu, J...construct for Antares is available through Addgene (Cat. # 74279). Enhanced Nano-Lanterns  Suzuki, K., et...wavelength above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed two imaging probes. One consists of NLuc...PMID: 27465542. PubMed Central PMCID: PMC4974478. 3. Schaub, F.X., et al. (2015) Fluorophore-NanoLuc BRET...et al (2). In this post, I’ll cover two great applications of the NLuc technology. The first couples NLuc...fluorescence. The paper reports a wide variety of applications of the Enhanced Nano-Lanterns including monitoring...2017) Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging...

  13. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...the stricter sequence requirement at the terminal 3’ PAM site (Fu et al. 2013; Pattanayak et al. 2013)...every CRISPR experiment. Off-target events due to >3 bp mismatches or that are sequence-independent are...PMID: 25566793. PubMed Central PMCID: PMC4349094. 3. Bell, Charles C., et al. "A high-throughput screening...off-target sites across the genome and pinpoint the location of mismatches based on the sequences of the genome... Figure 1: Schematic representations of the indicated unbiased genotyping options. Figure reproduced...those relating to DNA structure (e.g. epigenetic modification, bulges) that may also impact off-target edits...cell-based methods are genome-wide, unbiased identification of double-strand breaks (DSBs) evaluated by...

  15. 6 Tips for Grant Writing

    Type
    Blog Post
    Published
    Oct. 18, 2016, 2:30 p.m.
    ...alterations before submitting - handing them your application 3 days before the deadline isn’t going to be much...work and possibly a new perspective on the field. 3. Highlight your network and your collaborators It’...Series 4. Become a science communicator More and more fellowship applications are asking for a 200-300 ...good quality application. Even following this advice, only about 50% of my grant applications have been ...do need to be able to research and write your application, then give it to someone else to read and / or...it). It’s easy for reviewers to spot when an application is rushed (whether the idea isn’t properly researched...researched / developed or the application is full of typos) and these are unlikely to get past the initial...

  16. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    Published
    Sept. 17, 2015, 2:30 p.m.
    ...materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various steps ...doi: 10.1038/nprot.2011.413. PubMed PMID: 22157972. 3. Britt JP, et al. Use of channelrhodopsin for activation...There are several important considerations with fabrication of the fiber optics needed for optogenetics. ... for the implant depends on the dorsal/ventral location of your region of stimulation (the z-axis bregma...emitted from the end of the fiber optic implant scatters a great deal once it enters tissue, placement ...0.5mm above the site of your stimulation. The scatter effect is also an important consideration when ...Lasers can be purchased through Laserglow, which caters to optogenetics users and adjusts the lasers appropriately...

  17. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...knockout, activation, repression, and barcoding.  3) Are you trying to target every gene in the genome...sequencing. Representation - Most libraries contain 3-6 gRNAs per target gene, and maintaining the distribution... unique compared to the rest of the genome and located immediately 5’ to a protospacer adjacent motif ...coli, and T. gondii genes.  2) What genetic modification are you trying to make? Addgene carries CRISPR...next-generation sequencing to check the quality of your amplification. If you've obtained a ready-to-use lentiviral... As such, sequencing CRISPR libraries after amplification and after a screen requires the use of next-...experiment, the use of appropriate controls, multiple replicates and several cell types can strengthen your results...

  18. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually encodes...vessel size. Opting for a larger vessel, such as a 3 or 5-layer flask will require some initial optimization...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from...end-users, size of the transgene, and downstream applications should all be considered when choosing the correct... as resources available, cost and downstream applications. For example, while the PEG precipitation method...

  19. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ...expression (Sapozhnikov and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation...work well, and up to 10 base pairs away (especially 3’ of PAM) might work too.  High dCas9 and gRNA expression...approach (spoiler alert: it’s dCas9… with no modifications). Ready to get started?   Figure 1:... when DNMT1 methylates the genome during DNA replication, it doesn’t methylate sites blocked by dCas9....for dCas9 to interfere with DNMT1 even through replication. The dCas9 also needs to be removeable, otherwise...vivo using dCas9–peptide repeat and scFv–TET1 catalytic domain fusions. Nat Biotechnol 34, 1060–1065. ...

  20. Imaging Tools and Following a Passion for Basic Biology: Interview with Joachim Goεdhart

    Type
    Blog Post
    Published
    March 9, 2018, 2 p.m.
    ...passion and pay attention to the details. Chapter 3 - Answering interesting biological questions using...Twitter and has helped us update many of our educational resources. Listen to hear all about Professor...speeds science. Chapter 8 - The importance of communicating research findings through social media and other...
Showing: 181 - 200 of 466 results