...proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching an old antibody...on-target delivery of genetic cargo  By Alyssa NeuhausTo improve delivery and reduce off-target effects....1101/2023.02.07.527548.    Tracking cellular history with protein ticker tape By Brian O’Neill The Cohen...created a system for recording the transcriptional history of a cell. Lin et al. generated a fusion protein...ticker tape images could recover transcriptional histories, accurate to within 40 minutes. Figure ...their initial tests with thick slice (70 μm) immunohistochemistry (IHC) (Figure 5). PSD-95 is a scaffolding...

...control by Addgene before being distributed to customers. Details about our production protocols, titering...vivo studies. Preparations are then aliquoted and stored at -80 °C. Titer Titering is either performed by...preparations is assayed by comparing the relative stoichiometric ratios of the viral capsid proteins VP1, VP2...The preparations are then aliquoted, frozen, and stored at -80 °C. Concentrated lentiviral preparations...then resuspended in PBS, aliquoted, frozen, and stored at -80 °C. Titer All titering is performed on lentiviral... lentiviral preparations that have been stored at -80 °C and thawed. This ensures that any loss of titer...

...structures (at < 5 Å resolution) of the linker histone H1.8 in Xenopus nucleosomes and in complex with...has developed a new epigenetic editor, Coupled Histone tail for Autoinhibition Release of Methyltransferase...can be toxic, CHARM uses a cofactor fused to a histone tail to recruit and stimulate the endogenous methyltransferase...uses a fusion of the DNMT3L cofactor (D3L) with a histone tail (H3K4me0) to a DNA-binding protein (dCas9 ...team to develop self-silencing CHARM variants that stop their own expression after silencing their target... a familiar challenge for anyone who has done histology on brain tissue is identifying the cells you are...

.... We are the Customer Support Specialists here at Addgene. If you’ve called our customer service line ...'Regan Ordering jeans online from your favorite store is easy. Ordering genes on plasmids online is a ...really getting at one bigger question: How does a customer place an Addgene order using a PO number? Why ...full of information about how to deposit plasmids, store your plasmids or viral items, technical support,...

... Center for Regenerative Medicine of Boston University and Boston Medical Center. Special thanks to our...director of the Center for Regenerative Medicine at Boston University Medical School. Darrell’s research focuses...Chapter 1: Introduction to Darrell Kotton and the Boston University Center for Regenerative Medicine. Darrell...Center, and the Pulmonary outpatient clinic, all at Boston Medical Center. In addition he conducts basic stem...

...my training, I dove straight into my capstone project. A capstone project is a major project given to ...Figure 1: Addgene's summer interns presenting their capstone projects during the Summer Intern Poster Presentation... scientist. But I expected something wrong. My capstone project was based on plasmid cloning and I, the... Once everything was confirmed, I worked on my capstone project report and presentation, during the final...

...you questions. Remember that people like a good story and science involves characters – in this case you...or other places online, and consider how those stories are being told. Twitter can be a good way to follow...surprised to find that his friends and family finally understood why he cared to study mice on some distant island...the East Coast, there’s programs at both MIT and Boston University. At Addgene we're committed to scientific...sharing – both your plasmids and your research stories. Consider practicing communicating your research...

...resistance gene (missing the first two codons). Two stop codons are inserted between the truncated gene and...ligated into the MCS/SacI site in pUCXKT, removing the stop codons during backbone digestion and providing the...Toolkit for constructing fusion proteins with customizable N- and C-terminal modules   Thanks to the lab...Technologies. The MultiSite Gateway Kit is a customizable Gateway®-based toolkit that allows scientists...bound to the mRNA since they are tethered AFTER the stop codon. Thus a non-translated mRNA will appear yellow...: 10.1126/science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector...LacZ, puromycin or CreERT2 can be combined with custom-cloned vectors in a Multisite Gateway reaction ...

...IP and ChIP assays. Tag 96 genes at a time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein (csRNP...first step of optimization, we started off by using custom synthetic two-part cr/tracrRNAs for CRISPR experiments...tagging, pick a crRNA that cuts 8-15 bp downstream of Stop codon; for N-terminus tagging, a crRNA cutting 8...csRNP experiments, it can be ordered in bulk and stored in small aliquots at -80 oC. Assemble csRNP: Anneal...near the insertion site (within ~8-15 bp from the stop codon for C-terminus tagging) GC content of the ... hands, delivering Cas9 protein in complex with custom synthetic cr:tracrRNAs (csRNP) is the most efficient...

...not necessarily represent the views of Addgene. Boston, MA, USA 42.3601°N 71.0589°W – Alina Chan The ...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...to join the public conversation and to tell the stories behind our research.  That's why I marched for ...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...behind these two fingers the ninety-nine cubits of Aristotle, nor would you mention my small error and at the...second and that it is our duty to tell our science stories in a manner that encourages open discourse with...

...Emission Depletion (STED), and Stochastic Optical Reconstruction Microscopy (STORM). These dyes have specific...Throughout their history, fluorescent dyes have enabled the visualization of both organic tissues and...than was absorbed. This phenomenon is known as the Stokes Shift, or the difference in wavelength between ...technologies in fluorescent dyes/ligands Throughout the history of cell biology, fluorescent labeling techniques...fluorescent proteinFluorescent Proteins 101: A History of Fluorescent Proteins  ...

...skeptical reader of historical fiction (I can personally attest to this!). Following history from the Ice Age...Wrong - and the New Research That's Rewriting the Story by Angela Saini "This book is a smart and carefully...Strange & Mr. Norrell by Susanna Clarke "A quasi-historical/fantasy fiction. Two dueling magicians in the...of England by Edward Rutherfurd "A 10,000 year history of southern England. While the length may be intimidating...

... examples with plasmids deposited at Addgene.  PRESTO-TANGO: The ligands that activate many GPCRs are...Roth’s lab created an open source resource called PRESTO-TANGO (parallel receptorome expression and screening...transcriptional output - TANGO) (Kroeze et al., 2015). PRESTO-TANGO uses a modified Tango beta-arrestin recruitment...ligands for unknown GPCRs. You can find the entire PRESTO-TANGO kit at Addgene! TRUPATH: Bryan Roth’s lab...McCorvy JD, Giguère PM, Sciaky N, Roth BL (2015) PRESTO-Tango as an open-source resource for interrogation...

...initial reversal drive cannot restore the exact original sequence, it can restore the original protein-coding...are eliminated by natural selection, gene drives distort inheritance in their favor and consequently can...they should not be relied on exclusively because historical studies of pathogen research have conclusively...using a recoding strategy; a subsequent drive can restore the wild-type sequence (save for the residual sgRNAs...

...Fast-FT, and mK-GO. Large Stokes Shift (LSS): Stokes shift (named after George G. Stokes) is the shift in wavelength... most FPs, Stokes shift is less than 50nm (often much less).  For LSS proteins, the Stokes shift is ≥ ...

... wrong. It brought the whole field to a grinding stop and only now, 2 decades later, is it finally back...showing that AAV-mediated delivery of the RPE65 gene restores vision in the dog model of LCA (Lancelot, one ... 10 more years to obtain FDA approval and make history in the form of Luxturna™. A better vector for ... again 3. Acland G. et al., 2001. Gene therapy restores vision in a canine model of childhood blindness...PubMed PMID: 11326284. 4. Bennett J, 2017. Taking Stock of Retinal Gene Therapy: Looking Back and Moving...

...when balancing a handful of demanding courses, a capstone paper I really wanted to hit out of the park, .... I need it just as much in my current role as Customer Support and Operations Manager at Addgene. In ...about to keep silent and move on to other things…. Stop! Take a step back and think of a time you’ve lead...collaborator or co-worker and showed that you really understood their side of an issue. Maybe you also showed...article from The Muse. Kai Strickland is a former Customer Support and Operations Manager at Addgene and ...

...Ampicillin Ampicillin, often just called amp, is the historical king of the antibiotic resistance realm. Amp ... factor. Zeocin is handy because it can be a one-stop shop for selection across systems and species. The...entirely detoxified, even when Sh ble is expressed (Trastoy et al). Thus, it may not be a good choice for experiments...Hadley, W. K., & Brooks, G. F. (1986). Effect of storage temperature and pH on the stability of antimicrobial...11, 56. https://doi.org/10.1186/1475-2859-11-56 Trastoy, M. O., Defais, M., & Larminat, F. (2005). Resistance...

...yourself needing to brainstorm with a team. One major challenge with these brainstorming sessions is avoiding...with unlimited time? Another strategy for your brainstorm session is to start with something completely... ideas that I have found helpful in my team brainstorming sessions.  Allow the group time to think about...position or the longest tenure, but don’t let that stop you from speaking up.  If your ideas are not well...

...celebration, we thought it only fitting to share some stories of ways we’ve turned the available into the accessible...countries, import and export regulations, differing customs processes, and the distances the packages needed...other things, could ensure our packages navigated customs and arrived at their destination in a timely manner... to Design a Primer Creating Bacterial Glycerol Stocks Inoculating a Liquid Bacteria Culture Collections...scientists, from techs to grad students to professors, stopping by to say, “Everything I know about virus, I learned...