We narrowed to 433 results for: Hal
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TypeBlog Post... demethylase to demethylate enhancer regions (Mendenhall et al., 2013). By comparing gene activation when...sorting and selection of transduced cells. Grant Challen’s lab also created constitutive (pCMV-dCas9-D3A)...contributed to updating this article. References Haldeman JM, Conway AE, Arlotto ME, et al (2018) Creation...Fowler T, Rees R, Kramer A, Martens A, Edwards JR, Challen GA (2016) Reprogrammable CRISPR/Cas9-based system...Find plasmids from this publication at Addgene. Mendenhall EM, Williamson KE, Reyon D, Zou JY, Ram O, Joung...
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Technologies Enabled by NanoLuc® Luciferase
TypeBlog Post...information. Many of our technology platforms like HaloTag® Fusion Protein, codon-optimized Firefly luciferase...concentrations of biological molecules. FRET sensors face challenges of photobleaching, autofluorescence, and, in ...cyan-excitable donors, phototoxicity. Another challenge to using FRET sensors comes when employing optogenetic...Researchers have sought to alleviate many of these challenges by exchanging the fluorescent donor for a bioluminescent... analysis products at Promega. References 1. Hall, M.P., et al. (2012) Engineered luciferase reporter... -
Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands
TypeBlog Post...interchangeable labeling using the HaloTag® system, the new Janelia Fluor HaloTag Ligands expand the self-labeling... self-labeling proteins would be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme...Hayashi-Takanaka et al., 2014). Other dyes may prove challenging to stain intracellular compartments in in vivo...2017). These dyes were adapted into ligands for HaloTag® and are currently offered at Promega. Due to the... -
Our Most Popular Blog Posts from 2019
TypeBlog Post...and represent the diversity of tools we share on behalf of scientists. 9. Plasmids 101: Positive and Negative...fluorescent proteins at Addgene. 7. Designing Your Chalk Talk for the Academic Job Interview by Erik Snapp...blogger Erik Snapp breaks down how to prepare a chalk talk for an academic job interview including defining... research vision, organizing your vision into a chalk talk, and tips for practicing. 6. The Fluorescent... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...annurev-neuro-111020-100834 Challis, R.C., Ravindra Kumar, S., Chan, K.Y., Challis, C., Beadle, K., Jang, ...or lumbar spine. Image and caption courtesy of Challis et al., 2022. Where do they come from? Systemic...https://clover.caltech.edu. Figure adapted from Challis et al. 2022. Image created with BioRender.com. ...low doses a regional bias towards the cortex and thalamus emerges. AAV-PHP.eC effectively transduces neurons... Resources and references Recommended reading Challis R.C., Ravindra Kumar S., Chen X., Goertsen D., ...https://doi.org/10.1016/j.crneur.2023.100086. Challis, R. C., Ravindra Kumar, S., Chen, X., Goertsen,... -
Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools
TypeBlog Post...promoting domain. Each half was fused with one of the LOVTRAP domains. When both halves of the modified EB1...growth and extension. Now, they split EB1 into two halves (Fig. 1): one containing the microtubule binding...expansion of the genome editing toolkit. However, a challenge in optimizing base editing technologies across... -
Antibody Validation for Flow Cytometry
TypeBlog Post...antibody selectivity. Additionally, it can be a real challenge to troubleshoot RNAi failures. Potential explanations...cell survival or proliferation, an additional challenge is that only partial or ephemeral knockdown will...confirmation of antibody selectivity even more challenging. Correlation with RNA or Proteomic data from ...supportive of their specificity for the protein. A challenge to this approach is that the identity of the epitope...the specific sample and protocol. This can be a challenge to confirm. As such, the approach used may need... -
Plasmids 101: Blue-white Screening
TypeBlog Post...vectors encode a gene which, when expressed, is lethal to the cell. Cloning fragments are inserted into...MCS in the center of this gene, disrupting the lethality. This is similar to α-peptide DNA disruption in...positive colonies do have the plasmid containing the lethal gene. An example is the ccdB gene system commonly...auxotrophic cell lines), or genes which rescue a lethal phenotype. Resources on the Addgene Blog Read... -
Delivery Methods for Generating iPSCs
TypeBlog Post...be removed by re-expressing the transposase. A challenge of using PB to generate iPSCs is that genomic ...the cytoplasm independent of the cell cycle. A challenge of using Sendai is that since it’s replication...clones tested had plasmid integration. Another challenge is that multiple transfection makes it difficult...successful reprogramming. If cells contain molecular hallmarks of reprogramming, then they are typically assessed...induced pluripotency: Recent progress and future challenges." Journal of Cellular Physiology 228, no. 2 (... -
Transferable Skills Guide: Problem Solving
TypeBlog Post...matter of applying that framework to the myriad challenges we encounter. As you spend time in a lab learning...yourself if you missed any important details. Or challenge yourself to think of more creative solutions. ... know-how and more about how you approach new challenges. Know your strengths! Research experience...employer that you are well equipped to work through challenges from day one in your new position. Of course,... -
Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics
TypeBlog Post...each half with PhyB or PIF6, phytochrome-like fusion proteins from the plant Arabidopsis thaliana. In ...identical in sequence to the unmodified enzyme. Each half of the base editor is fused to a fast-splicing split...induces PhyBΔ and PIF6APB binding, bringing the two halves of Cre together. Image from Eisenhoffer et al.,... -
Gaining Leadership Skills Volunteering at a Professional Organization
TypeBlog Post... the Association for Research in Vision and Ophthalmology Research (ARVO). Hopefully sharing my experience...online and on monthly phone calls. This has its challenges as meeting in person is often much more productive... quarterly and annual reports I had to write on behalf of the Advocacy Committee would change format depending.... You just need to be prepared to take on the challenges and be proactive and engaged once you are in ... -
Transferable Skills Guide: Creativity
TypeBlog Post... needing to brainstorm with a team. One major challenge with these brainstorming sessions is avoiding ... review it. This survey generated 26 ideas, over half of which, when implemented, impacted the entire ...you can, set aside time to prepare for future challenges. This doesn’t have to be time-consuming or even... keeping your science going throughout these challenging times. Take a Break! If all else fails, one ... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog Post...cassette using tamoxifen-inducible Cre not only halted the neurological damage but completely reversed... Rett Syndrome, respectively. One of the main challenges facing Rett researchers is how to deliver the...adeno-associated viral (AAV) vector. The editase repaired half of the mutant MECP2 RNA in each of three neuronal... by Gail Mandel. There are numerous technical challenges to overcome for both of these approaches, such... -
What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program
TypeBlog Post...share all of these new organisms continues to be challenging. Organism specific viral vector systems would...thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community...proteins, and DNA. Euprymna scolopes (Joshua Rosenthal) The Hawaiian bobtail squid has an extreme number... a different protein. Did you know that most cephalopods die after they lay their 1 clutch of eggs? Propagation... -
Building Global Connections with the International Mentorship Program USA-EUROPE
TypeBlog Post...PhD, achieving our career goals can be a real challenge. On top of that, in the midst of a rapidly changing...mentees with a broader, first-hand vision of the challenges and career opportunities that truly exist outside...was much more than that. I discovered that the challenges that I am facing in my career are common problems... that many others have faced yet, still, this challenge did not hinder them from advancing their careers... -
Providing Plasmids to Researchers in Developing Countries: Addgene and Seeding Labs Team Up
TypeBlog Post...to face global challenges, we hope to empower those that need to face these challenges locally. We look...themselves on the front lines of emerging global challenges. However, these same researchers face significant... -
Addgene Stands with the Scientific Community
TypeBlog Post...and increased tariffs present an unprecedented challenge to the scientific research ecosystem, not just...years to come. We Are Here to Help Despite these challenges, Addgene remains steadfast in our mission to ...resilience and creativity in the face of funding challenges. By strengthening our commitment to collaboration... -
Your Top Requested Plasmid in 2015!
TypeBlog Post...only been available for a little over a year and a half, a small sampling of the research it has already...another great year of sharing! References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide... -
Fluorescent Proteins 101: History of Fluorescent Proteins
TypeBlog Post...after that, in 1994, Prasher’s collaborator Martin Chalfie expressed GFP in exogenous organisms (E. coli, ...and later C. elegans) for the first time. After Chalfie’s work, scientists really started to take notice...Nobel Prize in chemistry was awarded to Shimomura, Chalfie, and Tsien for their work on GFP. This great honor...