We narrowed to 911 results for: CAN

...constructs can be achieved by fluorescence-activated cell sorting, and stable gRNA expression can be achieved... But other enzymatic fusions, such as dCas9-TET, can, like the iron ball-and-chain that they resemble,...site is not as straightforward as expected. TET1 can have many effects independent of dCas9 targeting,...ve got some good news: complete DNA demethylation can be achieved by dCas9 alone, without TET (or any demethylase...demethylase) fusion. This may seem counterintuitive: how can a DNA-binding protein with no enzymatic activity ...DNA demethylation! With optimization, this method can produce nearly 100% demethylation without the confounds...block that unmethylated CpG so that nothing else can interact with it. Remember, the goal of this experiment...

...hybridoma cell lines can produce more than one mAb (Bradbury et al., 2018). Hybridomas can lose gene expression...into a fully functional antibody. These antibodies can be used just as you would use antibodies made from...purified antibodies from a patient’s blood), scientists can use mass spectrometry to determine the amino acid...from an established hybridoma line, the antibody can be converted into recombinant form through DNA sequencing...2010; Andrews et al., 2019). Finally, scientists can perform the entire process of antibody selection ...been cloned into an expression plasmid, the plasmid can be introduced into host cells, such as bacterial,...engineer them. Given the ease with which scientists can now improve genetically-encoded research tools, it...

... – Get your voice out there Here’s one thing you can do today: Sign up for a Twitter account. Check out...in local museums, zoos or botanical gardens. You can also work or volunteer for a science communication...you don’t have time to maintain your own blog, you can probably write for your institute’s blog. The Wellcome...Use your unique talent to communicate science. You can draw live sketches at conferences like @ATJCagan ... by taking a media or journalism course. Courses can help you gain work experience and build your network... expect them to be the “be-all-and-end-all”. You can, for example, get an MSc Science Communication or...involve the public in your research projects, you can set up a citizen science project. Answering the big...

...what facet of academia you are in, grant writing can be the lynch pin that results in your success or ..., and perhaps the most important bit of advice I can suggest, is to give yourself plenty of time when ...and why these findings will be worth knowing? This can sometimes be tricky, as most of our research builds...collaborate with specialists in different fields that can provide different insight into an area - not just...Furthermore, using new technologies to solve a problem can allow you to approach that problem in a novel way...results published in a paper, but positive outcomes can include poster presentations, talks about your work...grant application to your peers and collaborators can also help you get some critical insight into what...

... tools can distinguish between global strains of the Zika virus (Asian-, African- and American-lineages...monitor and slow this spread until vaccine programs can be put in place to provide community protection. ...allow for the rational design of RNA sensors that can detect virtually any RNA sequence (Green et al, 2014... later. The Zika publication timeline I think we can speak for all the authors on the paper when we say... of a rapid design and prototyping pipeline that can take pathogen sequence information from public databases...technology on active virus, as well as shown that we can detect the virus in a clinically relevant setting...patient blood and urine samples; we showed that we can detect the virus directly from infected monkey blood...

...Experiments using enzymatic labeling can be difficult to quantify accurately, but can be especially helpful to generate...imaging, repeating for each target. Alternatively, you can perform simultaneous detection with a mixture of ...secondary has a different color fluorophore that you can image separately on your microscope. Abcam's sample...important consideration when deciding whether you can do simultaneous or sequential labeling is what conjugated... fluorophores, then labeling with the other, you can still record the signal separately for each target...specificity, and dye meets your needs and budget, or you can make your own. For example, to label the rat brain... automatically from the microscope software, you can always recolor them! In Fiji, for example, you’d ...

...replication can involve an RNA or dsDNA intermediary, depending on the class of virus. RNA genomes can be transcribed...removed. This means the vector can deliver a genetic payload to a cell but cannot create new particles. As ...have a wide variety of possible genome types that can be either double stranded (ds) or single stranded...Image credit: Louten, 2016 Viruses with DNA genomes can either follow central dogma (gDNA to RNA to proteins...were able to create far safer viral vectors that can be used in BSL-1 or BSL-2 labs. Viral vectors are...shell (Taylor, 2014). They’re so simple that we can’t even decide if they’re alive or not. Yet these simple...

...as yeast. HEKs can also grow in suspension, which allows for large scalability, and can also be adapted...cells can catch a cold—or worse! HEK-293T and its derivatives are immortalized, but cell health can degrade...reproducible results. It’s also widely available, which can make for a seamless transition of biological tools...degrade over time in culture. This can affect factors that could skew research results (gene expression... tumorigenic, which makes them a candidate cell line model for cancer-specific research. Membrane receptors...adapted for serum-free growth, which significantly reduces the cost of production. Research applications ...

...include CRISPR machinery (which can be supplied separately). Libraries can target specific pathways or the...lineage tracing, among other applications. Cells can be identified using fluorescence, like in the pMuSIC...that prevent their expression. The knockout cells can be used to identify genes that underlie a particular...CRISPR isn’t the only game in town! Pooled libraries can be designed for all kinds of different high-throughput...with a pooled library. Surface display screening can also be conducted with yeast. Created with BioRender.com...

...information so we can follow up with you if we have any questions and so that we can give you recognition...experimental details in research articles, scientists can use the AAV Data Hub to quickly find details they...next experiments using a specific AAV tool. They can search the database by certain features like serotype... the community. There are a few simple steps you can take to input your data for sharing on the site. ...injection and detection methods. For example, you can provide location, titer dose (Example: 1E13 GC/mL...

...semi-synthetic or fully synthetic compounds. Antibiotics can be categorized based on whether they directly kill...bit of a gray area as some bacteriostatic reagents can kill bacteria when used at high concentrations (and...antibiotic stocks, see Addgene's Reference Page. How else can antibiotics be used in the lab? Historically, antibiotics... for plasmid selection. Additionally, scientists can use the loss of resistance as a marker for successful...Carbenicillin is more stable than Ampicillin and can be used in place of Ampicillin in most applications... Simplify Your Cloning Experiments See Where You Can Find Antibiotic Resistance on Our Plasmid Pages ...

...email to get information if you can help it. Observe as much as you can in person. Rotate in the lab, if...Connie Cepko to learn about her mentoring style. How can I find out if an advisor is a good mentor? You have...for scientists to change labs – do as much as you can to ensure a good fit in advance. When a lab is interviewing... – I am different.”  Unfortunately, a bad mentor can be bad news for anyone. Try to speak to them more...excellent mentors out there. With some good pre-work you can find a lab that will deserve your hard work and an...

...dual nickases to generate a knockout or edit. This can help reduce off-target activity. Deleting a large... multiple genes at once. Using multiplexed gRNAs can target multiple locations in the genome to modify...common question Addgene Senior Scientists receive is: can I express more than one gRNA from a single promoter...plasmid is available from the Bullock lab, and gRNAs can be inserted using Gibson Assembly or SLIC cloning...sequence, creating flanking overhangs. These overhangs can be customized to link together multiple fragments...IIS enzyme, the unique flanking overhang sequences can link together to allow for ordered assembly into ...Lentiviral Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing wt...

...cleavage, the fluorescence is no longer quenched and can be visually detected using lateral flow strips. Since...technique Sensitivity: Single molecule per milliliter can be detected – in the zeptomolar range Time: ~1 hour...Cleavage results in fluorescence and this approach can be incorporated into a lateral-flow readout slip ...Protospacer Adjacent Motif) for target cleavage. This test can be performed in two steps for challenging samples...the ssDNA-FQ reporter. The resulting fluorescence can be observed under either no excitation light, UV ...field-ready, Bluetooth-enabled PCR machines that can even be battery operated, made accessible by the ...sequences. Upon binding, changes in fluorescence can be visualised. Another approach was to apply the ...

...measured via the light produced. A similar approach can be used to analyze cell signaling pathways or miRNA...have other advantageous properties. This approach can improve live-cell imaging by avoiding autofluorescence...luciferase subunit to other proteins, researchers can test whether those proteins interact, since light...luciferase components together. A similar approach can test protein-protein interactions using BRET. A luciferase...Considerations for luciferase assays Luciferases can only produce light in the presence of a compatible...however, secreted luciferase or alternative substrates can permit measurements of luminescence from live cells...Luciferase variants with incompatible substrates can also allow multiplexing with experiments that supply...

...was successful in your organism or cell type? You can use DNA sequencing or other molecular cloning techniques...without any off target effects. However, these methods can be labor intensive and quite time consuming.   As...activity.  By using the RFP:GFP ratio, edited cells can be sorted from unedited cells. In an experiment that... once a mutant is identified, the PB transposase can easily and seamlessly remove the integrated GFP reporter...fluorescent protein (RFP), named GEMCherry2, that can rapidly and efficiently evaluate Cas9 cutting sites...genomic region inserted into the N-terminus of mCherry can be targeted by a sgRNA guided Cas9 causing a DSB ...depending on the nucleotides inserted or deleted, can result in an in-frame shift of mCherry and thus expression...

...But base editors aren’t perfect. They may be slow, can only target certain sites, or make only a subset ...with a faster deamination rate so that deamination can happen before Cas homologs let go. The Liu lab developed...editor, a functional gene III product is produced and can infect bacterial cells. If a phage does not deliver... The editing speed dictates which phage variants can propagate faster than the rate of dilution (passage...editor was named ABE8e. ABE8e is not only fast, it can also be multiplexed: when tasked to correct two point...mutations. Together, cytosine and adenine base editors can correct all four of these kinds of point mutations...of ABE8e to the base editing toolbox, scientists can now make these edits much more efficiently and in...

...GPCR signaling can also modify other properties such as endogenous gene expression, which can lead to changes...chemogenetic receptors hijack cell signaling pathways and can lead to such responses as decreased heart rate (Redfern... is important to consider that the same receptor can elicit different responses depending on what cell...manipulation are neurons where chemogenetic receptors can be used to alter excitability by lowering or raising...). While some scientists argue that these issues can be mitigated by using low doses and running appropriate...such as C21, CLZ, Perlapine, and Olanzapine, that can also activate muscarinic-derived DREADDs. A ligand... of off target effects. These synthetic agonists can penetrate the brain and some, such as varenicline...

...link genotype to phenotype that can be used with mutational scanning: ligand binding assays, phage display...saturation mutagenesis Several methods can be used to create mutational scanning libraries. While each of these...saturation mutagenesis as a quick and easy technique that can be used to generate complex libraries of mutant plasmids...thousands of protein variants can be tested in one experiment. There are many functional assays that link... choosing and validating a selection system. You can test the effect of multiple mutations in combination... bases (NNN) at any given codon site of interest can be cloned into an expression vector with Gibson assembly...mutagenesis FAQ What’s the maximum length of codons that can be mutated? 88 codons, or 264 basepairs, was the ...

... happen before you even receive the product? How can we consider the environmental impact of those processes... waste directly into the oceans.As consumers, we can choose products that are better for the environment...sacrificing performance or quality. To do this, we can use the ACT label, which is an eco-nutrition label...sciences, some of these plastics are high grade and can be washed and reused. This might sound risky in terms... incentives (for example, by grant agencies), we can shift toward a more sustainable one. The math on ...