...observe what’s happening inside cells for at least a couple of decades. Green is the classic color, but fluorescent...signal-to-background ratio, thus, allowing higher resolution in samples containing substantial autofluorescence background...direction of biosensors capable of revealing, for example, how muscles work inside a living animal. For those...

...CRISPR Antimicrobials Day 7: Multiplexing and Cas9 variants: CRISPR 101: Multiplex Expression of gRNAs and ...Cpf1: A New Tool for CRISPR Genome Editing and Multiplex Genome Editing with CRISPR-Cpf1 Day 4: Edit validation...

...kind of walking them through the steps of, for example, cutting open a heart to look at the inside. We...a lot of interactives and videos, as well, to supplement the hands-on [aspect of the labs],” says Caitlin...new technologies in education and ask ‘Can we implement that?’ [And think about] where we [eScience] are...

...Iowa in the Department of Pharmacology, I taught a couple of lectures in the undergraduate pharmacology course... editing sphere.” But things aren’t always that simple, especially when your spouse is also in science... stumbled upon eScience by luck, by chance, by complete happenstance. It was on Indeed or one of the job...

...helps improve your interpersonal relationships with people you’re collaborating with remotely. Depending on...as Slack. Share data and documents While having multiple scientists edit the same document may sound like...collaborators. Being able to present data from multiple aspects of a collaboration and immediately discuss...

...University of Akron, Ohio. Congratulations! There was plenty of geeky fun to be had at ASM Microbe as well as...science theme, the Artwalk on the exhibit floor had plenty of wonderfully nerdy art for sale, and photos of...presented at the conference, it looks like there are plenty more exciting plasmid tools on the horizon for ...

...skills help when it comes to finding a job. Many people in science will work in industry or take on collaborative...and their interest in science more generally to people outside of academia. Here are my recommendations... they start asking you questions. Remember that people like a good story and science involves characters...

...4 of the Cong, et. al, 2013 Science paper. For example, if you want to use double nickase, you could express...selection marker to select transfected cells (for example, plasmid with a puromycin resistance cassette, ...individual genotyping (using Sanger sequencing, for example) on each colony in order to screen for positive...

...common lexicon for genome editing studies. These complementary goals are aimed at improving translation of ... loosely paraphrased in the tweet below. The complexity and challenges of homology directed repair (HDR...editing and low off-target editing rates across multiple systems. Read our introductory blog post, then...

...an ITR during routine plasmid propagation, with complete ITR loss abolishing AAV packaging. To prevent ... like as long as it’s ~4.4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas...also drive different levels of expression. For example, CAG is a strong promoter that drives very high...

...Balazs. “By combining those with [sera] we got from people that have been given the vaccine, we've been able...used live virus (Cele et al., 2021). “We weren't completely shocked [with these results] because we were ... AG, Iafrate AJ, Naranbhai V, Balazs AB (2021) Multiple SARS-CoV-2 variants escape neutralization by vaccine-induced...

...jGCaMP7 sensors with different properties (for example, overall brightness or on/off kinetics) that makes...quantify targeting of particular cell types. For example, the mDlx enhancer restricts expression to GABAergic... particular catalog item is available for order please email [email protected] with the vector name and...

...incorporate into their own labs. This week we are pleased to announce the submission of the results of our...be examined. Increased yields with sucrose supplementation is serotype specific In our study we observed...preparations for, we wanted to be certain that this supplementation strategy would have no effect on the purity...

...ever like to write about a recent plasmid deposit please sign up here.   Listen this episode of Hot Plasmids...APOBEC3A-Cas9n-UGI and APOBEC3B-Cas9n-UGI base editing complexes are more efficient than the BE3 plasmid containing...exhibiting its versatile utility in quantifying multiple gene editing activities. Click here to find the...

...manipulated with mutations in the replicon. For example, the pMB1 ori maintains about 20 copies per cell... any questions? Need something clarified? If so please, post in the comments.  References Chatterjee A...Schultz PG. A versatile platform for single- and multiple-unnatural amino acid mutagenesis in Escherichia...

...many different things in parallel.” Alternating Hemiplegia of Childhood Melodi McSweeney (right), a graduate...responsible for the disease known as Alternating Hemiplegia of Childhood (AHC). Patients affected by AHC ...from recurrent attacks of unilateral paralysis (hemiplegia) that are often painful and accompanied by migraines...

...discussion. The Forced Choice Analysis is a fun example and you can find it in our Mentoring for Scientists...help the Mentee feel a sense of accountability to complete action items and make progress. To illustrate ...with regular accountability checks. Focus on implementation of your goals and respect the time your Mentor...

...about FPs than is currently represented at fpvis (complete spectral information, accession numbers, sequences...the goal of encouraging user contributions (using simple web forms) while also maintaining a relatively ...in the early stages of development, and there is plenty of room for improvement and elaboration.  Current...

... SARS-CoV-2 interacts with human proteins. For example, we used HEK293T cells in our first study, but ...collaborative science, and making it accessible to all people, not just scientists! *If you are requesting plasmids...plasmids for coronavirus or COVID-19 research, please email Addgene at [email protected] with your order...

...ever like to write about a recent plasmid deposit please sign up here.        A retroviral mouse CRISPR...Thus, the bEFPs bind the UAS via the dCas9-gRNA complex which allows transcription from the promoter.  ...should serve as both a powerful tool for generating complex regulatory systems and further the study of natural...

... It doesn’t have to be this way. We can never completely eliminate the risk that a minor variation in ...collective priority list. There has been progress—for example, a number of funders (e.g., the NIH) now look for...Press.  She trained originally in cell biology, completing her PhD at the MRC LMB in Cambridge UK and then...

...size 20 µL samples of control AAV that scientists can use for serotype testing. These samples are available... particular catalog item is available for order please email [email protected] with the vector name and...

...produced? Purifying exo-AAVs is actually a bit simpler than the classical purification protocol involving...have tried to modify it to improve yields. For example, transfecting the producer cell line with an expression...were always normalized to their genomic titers). Examples include the use of exo-AAV1 for transducing sensory...

...techniques are continually being refined; it is completely plausible that scientists will eventually have...many things we can navel-gaze over from the last couple of weeks, including whether or not scientists can...history of medical research, there have always been people who unethically experiment on humans, and they ...

...ever like to write about a recent plasmid deposit please sign up here.        New toolkit for N or C terminal...binds to TIR1 of the SCF-TIR1 ubiquitin ligase complex, promoting its interaction with the degron sequence...used ChRs in that they cause a rapid and almost complete desensitization when exposed to continuous bright...

...variants. Image adapted from Aggarwal et al…, Supplementary Figure 3.   Figure 3: pH titration...variants. Image adapted from Aggarwal et al…, Supplementary Figure 2.   Improved trafficking and localization... Multiple Authors...

... most familiar bioluminescent organism for most people is the firefly (Photinus pyralis) and perhaps not...luciferase activity as a measure of signaling activity. Multiple response elements are cloned into a plasmid upstream... will result in luciferase expression and more complex cell treatments can be designed to further explore...

... enzymes, some of which are also functional in multiple species (read Arbor Biotechnologies' press release...nt direct repeat) mediating potent knockdown. Multiplexing from the CRISPR array yielded >90% knockdown... necessary. Konermann et al. compared CasRx to multiple RNA regulating methods: small hairpin RNA interference...

...For safety and production reasons, multiple plasmids with many components are used to package lab grown...with each other to allow DNA synthesis of the complementary strand to yield dsDNA. Unlike LTRs, ITRs are...): Cold Spring Harbor Laboratory Press; 1997. Principles of Retroviral Vector Design. Available from: ...

...researchers new to GCE aren’t able to successfully complete their experiments and many others are dubious ...universal techniques that researchers can easily implement in their own labs. They provide resources, knowledge...bulletin board on their website specifically so people can talk about GCE. In fact, their first protocol...

...cool because they can transmit information, for example, from one of the 180 regions to another or from...even further defined population of neurons. For example, say I wanted to access all the neurons that signaled... Multiple Authors...

...shRNAs are short RNA sequences which target complementary mRNA sequence for degradation. Viral introduction...targeting an exon in your gene of interest, for example. Some customization is required, but many viral...them an ideal delivery system for human patients. Multiple viral-based gene therapies have already been approved...

... look at how we create this content, using the example of our Antibody Guide, which was published alongside...can. We even have a Slack channel just for asking people if they can write or review things and another ...require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in the past few ...

...of both AsLOV2 and the nanobody. As a proof-of-principle the Toettcher Lab monitored how AsLOV2 insertion...manipulation of the signaling pathway.  As a proof-of-principle, the team created an OptoNB to control the Ras...levers to pull to control signaling pathways. For example, if the OptoNB could bind and block a receptor ...

...specific region of your protein of interest. For example, if you are studying a protein that may bind a ...recognize the protein even though it is present in your sample. Additionally, there is a lower likelihood that...higher than that of polyclonal antibodies. For example, it takes longer to develop a hybridoma than it...

...agriculture, are now synonymous with GM to the point that people cannot imagine any use for GM technology other ... is not food, not medicine, and not mandatory – people can decide for themselves if they want this beautiful...developed a genetic tool for plant engineering, please share it with Addgene to both build the plant synthetic...

...based on size. Whether higher molecular weight complexes are detected will depend on the homodimer affinity...will be problem-free. In figure 1 we provide an example of the OSER assay with a number of fluorescent ...tested. To conclude, we recommend that you try a couple of different fluorescent proteins when protein ...

... practice to compare degradation rates between samples treated with or without an autophagy inhibitor....accumulates in the vacuole. See Figure 2 for an example of what fluorescence looks like for cytoplasmic...Fluorescent Protein Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy...

... the LoxP sites, which is more difficult than a simple knock out. Looking for some permanent knock out...transcripts. When transiently introduced, they can deplete endogenous protein levels for 1-3 days. There’... tunable control These systems can be easily implemented for exogenously introduced proteins, as many ...

...ever like to write about a recent plasmid deposit please sign up here.        AAV delivery of split base... Type IIS restriction sites to easily assemble multiple DNA fragments. This system has been used to construct...controlled by Cre recombination. Each cell can have multiple copies of the transgene that recombine independently...

... further demonstrated that PRIME-del can be “multiplexed” using pooled paired pegRNAs.  Read the paper...Xue’s lab also uses paired pegRNAs targeting complementary DNA strands to program large genomic deletions... without small insertions. But, instead, PEDAR couples this deletion process to a prime editing enzyme...

...affect it. While tumors may be the stars, there's plenty of action outside of tumor cells, and recent studies...deeply connected to these 3 stages and contains multiple cell types: fibroblasts, myofibroblasts, neuroendocrine...spread and drug resistance? Although we’ve seen how multiple types of cells affect tumor progression, enzymes...

...intensive to process a large number of microbiome samples. To enable large-scale, longitudinal studies of...and can approximate mRNA concentration within a sample. Using this system, the Collins lab was able to...lab validated their platform on clinical stool samples by comparison to qPCR and demonstrated that it ...

...preserves RNA structure and naturally occurring complexes present in the cell, it can also introduce non-physiological...non-physiological RNA-protein interactions due to sample processing. Require prior knowledge that the protein...levels seen in HHCS patients. These are just three examples of how RaPID can be used to study RNA-protein ...

...below) Because off-target sequences have less complementarity to the gRNA, they have a higher propensity ...for their on-target and off-target activity at multiple genomic locations in HEK cells. Further testing... Hiroshi, et al. "Crystal structure of Cas9 in complex with guide RNA and target DNA." Cell 156.5 (2014...

...erlotinib. Looking at barcode abundances among multiple replicate populations, they found that similar...went on to show that combination treatment with multiple drugs reveals even smaller preexisting populations...dynamics in response to cancer therapy using high-complexity barcoding." Nature medicine 21.5 (2015): 440-...

...have all heard. It’s a truth that extends to both people and the tools we use. Scientists utilize an array... to stop functioning properly or be something completely different than what was originally intended. ...Addgene receives deposited plasmids, they undergo complete plasmid sequencing using an in-house Illumina ...

...achieved with a 30-minute incubation, even with multiplexed guides. Figure 1: A) Cell-penetrating ...generated a series of StayGold mutants and identified a triple mutant, Y187F, R144I and T155Q, which they named... Multiple Authors...

...viruses that can infect people, cells, and animals. Some of these viruses are completely inert, but others ...protection are key to lab safety and the same principles apply to viral handling. Gloves are always required...

...others are studying and gives you a sense of what people are excited about. Following scientists in fields...more about authors by exploring author pages (for example, Melina Fan’s author page). These pages show all... Key features Setting up a personalized feed is simple. You can enter as many search terms as you like...