...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...

...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 (...

...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,...

...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.,...

... 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 ...

... 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 ...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

... some challenges during my internship, but nothing could have prepared me that those challenges would ...exploring my different interests but also a way of challenging what I am used to. Brandon Solis, marketing...

...possible before,” said the Broad Institute's Ophir Shalem. “Whole regions of the genome which were not accessible...depends on your experiment and method of selection. Shalem and Sanjana say they kept a high representation... CRISPR-Cas9 Knockout Screening in Human Cells. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson...

...cycle of one step. Challenges in the LEGO science video process One major challenge when we started was...will bring people together and make these daily challenges a little easier to overcome. We want to encourage...

...researchers involved. Communication is a major challenge when researchers are communicating across disciplines...disciplines or doing different tasks. But these challenges can be exaggerated when communicating across ... help other teams successfully overcome these challenges, too. Schedule regular meetings Scheduling a...

...team called Running for Rare Disease that runs on behalf of the National Organization for Rare Disorders...HOPE; a fitting superpower for those running on behalf of rare disease sufferers like AJ. As the Blue ... First-In-Man studies. Kurt has been running on behalf of AJ Gonzales, a young boy suffering from FOP,...

...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs ...tools for studying translation in living cells. Halstead, et al. Science. 2015 Mar 20;347(6228):1367-671...genes involved in neurogenesis, which had proven challenging using the existing dCas9-VP64 activator. The ...Designing customized fluorescent reporters can be challenging enough, but incorporating multiple reporters ...tools for all-optical electrophysiology A major challenge in the field of all-optical electrophysiology ...destabilizing sequences were added to shorten the half-life of both the luciferase mRNA (3’UTR AUUUA [5...

...translation inhibitors cycloheximide and puromycin, Halstead et al. verified that translation is necessary ...translation is known to be important in development, and Halstead et al. used TRICK to examine translation of Drosophila...Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor...