... nuclease domain. These mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 ...the next 3 ranked sgRNAs. Dolcetto was able to discriminate between non-essential and essential genes similarly... an improved activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA aptamers, ... active sgRNAs for CRISPR-Cas9–mediated gene inactivation." Nature biotechnology 32.12 (2014): 1262. PubMed...

...proteins of interest has occurred. Although the original Y2H systems utilized the yeast Gal4 activator,...for screening greater numbers, but the latter eliminates the step of identifying prey proteins after screening... the experiment to reduce the likelihood of indiscriminate reporter gene activation. Including a prey-... libraries can be screened using both N- and C-terminal fusions of these proteins. This way both ‘ends...

... bit of Addgene history... As you may be aware, Melina Fan, her brother Ken Fan, and her husband Benjie... corn toss boards, Kan Jam, and Ladder Ball (I finally discovered my hidden talent in Ladder Ball... there...popularly- cookies). 3. Do something outside of your ordinary environment Team building events like Escape the...Veckerelli is the Administrative and Operations Coordinator at Addgene. In addition to her operational responsibilities...

...last month it’s been viewed by users in the US, China, India, Canada, the UK, South Korea, and beyond!...protocols So that’s great, but what if someone in China improved upon that protocol? How would the thousands...prior to publication, and this will delay the dissemination of your improved protocol. Many scientists, ...need for quicker, more effective updating and dissemination of experimental protocols, simultaneously allowing...

...suitable partner, connect the relevant teams, coordinate implementation, and now must focus on managing...some microscopy enthusiasts to start a microscopy seminar to connect the imaging community and to create ...university approachable, friendly, and helpful. Finally, don’t forget about collaborations and projects...for lunch, make a point of talking to them at a seminars, or even set up regular meetings. Meeting people...

...exposing you to relevant researchers in the field. Finally, you can sound credible and cool when you casually...research papers on these particular themes, for seminal papers on these themes, and also for articles from...citations so that people can go back and read the original reference for the data. For example, we summarized...how big the studies were, and you could find the original paper if you wanted to learn more. Another example...

... Originally published Jul 14, 2015 and last updated Sep 16, 2020 by Beth Kenkel.  CRISPR genome editing...catalytically dead dCas13b to the catalytic domain of RNA deaminase ADAR2. Constructs containing the ADAR2 truncation...orthologs works for some applications, it doesn’t eliminate the need to deliver larger cargo such as base ...Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated...

...Microbe meeting, but it was my favorite because I finally felt like I had a network of people to talk to ..., Kat Milligan-Myhre, and Pragati Chengappa.  Financial and moral supporters Denise Akob, Sara Alexander...Heather Hendrickson, Jonathan Jacobs, Mo Kaze, Christina Kellogg, Heather Martinez, Pat Schloss, Tara Smith...McGovern, and Julie Wolf. Apologies to anyone I’ve inadvertently left off this list. Many thanks to our guest...

... in directly controlling muscle activity and coordinating movement, several additional brain areas that...dark areas (protected) as opposed to brightly illuminated ones (risky). They also prefer to explore novel... (Howerton at al. 2014). Addgene has several recombinase-expressing ready-made AAV vectors which can be...scientists study such as sleep, food intake, mobility/coordination, learning, and memory. We’ll cover these in ...

...strain distribution centers in the US, Europe, and China. In addition, the Zebrafish Model Organism Database...screens. Microscopy-based lineage tracing can help illuminate how signaling pathways—such as those regulated...patterns of gene activity and cell divisions that coordinate development of the body plan (Farrell et al.,...transgenesis with Tol2 and application for Cre/lox recombination experiments." Methods in cell biology. Vol. ...

...this is where most of our conference room names originated from. Let’s take a look at some of the stories...known to play a role in neural development, sex determination, and anterior-posterior segmentation (Jennings... SB transposon system including those from the original publication, Ivics et al 1997. Spaghetti Squash...may think that a gene called "spaghetti squash" originated from the squash itself, but think again. Spaghetti...

...their best interest to do so, sure, but it also eliminates that feeling of floundering one can have when...also need to talk to Tony from legal, Diana in finance, and Wu in graphic design. You could dread these...career options are expanding. As someone who coordinates cross-team projects at Addgene, I find that my... that led you into science in the first place. Finally, if you still find the idea of not holding a pipette...

... a second recombination event between the two flanking repeats. In cells where recombination has not occured... serves as a counterselectable marker in the destination vector. Successful cloning occurs when the ccdB...6308648. PubMed Central PMCID: PMC384129. Aizenman, Einat, Hanna Engelberg-Kulka, and Gad Glaser. "An Escherichia...

...but some applications have been challenging or inaccessible. The Ting lab recently made two deposits that... Figure 3: A) LOV-Turbo becomes active when illuminated with blue light, catalyzing the biotinylation...that had been trafficked from the “source” to “destination”. Images adapted from Lee and Cheah et al. 2023.... (2019). A toolbox of IgG subclass-switched recombinant monoclonal antibodies for enhanced multiplex ...

..., homology dependent repair (HR, homologous recombination; SSA, single strand annealing) took place and... with RNAs encoding Cas9 into the oocyte. This original method was developed as the use of CRISPR/Cas9... RNA and therefore there has less chance of contamination with RNase leading to degradation that can set...pups. Taking a toe, a finger, a tail tip, or a combination of these will allow you to determine which pup...

...circuit, while a nose-poke at a separate hole (inactive hole) does not. If activation of the circuit is...unwanted phenomenon or it could reveal something fascinating. Behavior is based primarily on careful observation...experimenting and hopefully optogenetics will help illuminate (is that pun worn out yet?) your research. Derek...PMCID: PMC4165888. 6.Witten, Ilana B., et al. "Recombinase-driver rat lines: tools, techniques, and optogenetic...

... practice, the term "promoter" describes the combination of the promoter (RNA polymerase binding site)...synthetic promoters, which typically include some combination of other promoter elements, and tend to be more...expression mRNA Mammalian promoter from phosphoglycerate kinase gene.  Constitutive  Widespread expression, but...optogenetics mRNA  Ca2+/calmodulin-dependent protein kinase II promoter Specific Used for neuronal/CNS expression...

...To acquire these measurements, the cytometer illuminates the passing cell with a laser and detects the...colleagues.   Figure 1: The laser pulse illuminates the cell and is scattered forward at a low angle...represents the time and intensity of the cell’s illumination (see Figure 2). H describes the maximum signal...For our last example plot, I will show you a combination of two stains, in this case through conjugated...

...This post was originally written by Melina Fan and updated Nov 3, 2022 by Susanna Stroik. You’ve designed...outlined below, the technique can be used as a preliminary validation on your bulk population. This way ...TIDER (Tracking of insertions, deletions, and recombination events). TIDER can screen for specific homology-directed...

...the correct size, and purify the DNA fragments. Finally, you simply incubate these three fragments together...loop (as might be anticipated to occur within a terminator sequence), since this would directly compete ...one step with no scar sequences remaining in the final product. The long overlapping region between fragments...different fragments. One way around this is to use a combination of standard sequences with overlap regions, such...