...facilitating recovery of edited animals. Figure 1: In the background are 504 restriction digests... demonstrated that selection for repair of a temperature-sensitive pha-1 mutation significantly enriches...making screening for knock-ins much more efficient. Pure PCR repair templates are critical for efficient ...editing efficiency improved by using more dilute mixtures (Doksin et al., 2018). The Mello lab also reported...visit Addgene’s C. elegans CRISPR plasmids page. Future perspectives It is interesting to note that recipients...and PCR-based homologous recombination donors in cultured Drosophila cells. Nucleic Acids Research 42:e89...

...can be tricky, and it takes extra effort to make sure everyone in the meeting has an equal opportunity...emailing documents to one person at a time. Just be sure there is a point person responsible for incorporating...will come from the collaboration. Some things to figure out are timelines for experiments and manuscripts...from each other, access specialized equipment, or ensure that techniques are being performed the same way...feedback with everyone involved can speed up the future directions of the project. The state of research...

...discoveries, the world of academia often emphasizes structured methodologies and rigorous research plans. However...strategy could entail immersing yourself in diverse literature. By delving into new topics and expanding your...taking field trips, and spending time outdoors in nature or visiting museums can be excellent methods to... times (or use the notes app on your phone) to capture fleeting thoughts and connections that emerge during...recognizes marvel in all things.   Conclusion  In the structured world of lab research and PhD studies, where ...

...molecular biology toolbox organized, today’s post features a list of free online molecular biology tools ...provides more tools for plasmid mapping and design (Figure 1). Benchling: While you might think of Benchling...Wayne Davis from the University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame...tools Addgene's analyze sequence tool: Annotates features, BLASTs, aligns, digests, and translates a sequence...access this tool, set up an IDT account. Figure 2: Addgene's Analyze Sequence tool.   DNA sequence...

...invading phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins... immune system seems like the winning ticket to ensure prokaryotic survival; however, viruses, too, are...strongest will to survive. Genome engineering and future directions CRISPR/Cas inspired genome editing was..., Mojica FJM, Wolf YI, Yakunin JvdO, Koonin EV. Nature Reviews Microbiology. 2011 Jun; 9, 467-477. Bondy-Denomy...Bondy-Denomy J, Pawluk A, Maxwell KL, Davidson AR.  Nature. 2013 Jan; 493, 429–432. Cong L, Ran FA, Cox ...

...because it turns a dynamic process into a static measurement and it provides limited information about what...fluorescent autophagy reporters now allows for the measurement of autophagic flux, or the changes in autophagic...strength excited at 586 nm to 440 nm serves as a measure of autophagy induction: when Keima is in the cytosol...is high. When untargeted, Keima can be used to measure bulk autophagy, but the addition of a mitochondria-targeting...18094608. Sargsyan, A., et al. "Rapid parallel measurements of macroautophagy and mitophagy in mammalian...

...2010). Rather than being released into the cell culture supernatant, the viral particles associate with.... Coroadinha, A. S., et al. "Effect of osmotic pressure on the production of retroviral vectors: Enhancement...overproduction of antibodies by applying osmotic pressure and sodium butyrate." Biotechnology and bioengineering...Shen, Chun Fang, and Amine Kamen. "Hyperosmotic pressure on HEK 293 cells during the growth phase, but ...serotype-dependent release of functional vector into the culture medium during adeno-associated virus manufacturing...

...we've been able to measure their relative ability to neutralize.”   Figure 2: Sera used in the...was not tested at the time of this study. Figure 1: Neutralization levels against spike pseudovirus...mutations was surprising,” says Balazs.   Figure 3: Neutralization outcomes of convalescent sera....V2 from neutralization by convalescent plasma. Nature 593:142–146. https://doi.org/10.1038/s41586-021...

...🎶 “Bye bye L-arabinose drivePut your cultures in the shakerTurn the LEDs onAnd when you wantYou can ... a light-responsive domain that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible...light-inducible dimerization domain, Vaucheria frigida aureochrome1 (VfAu1), led to functional BLADE variants, too...expression needed for most applications.   Figure 1: With blue light, the VVD domains of the BLADE...how spatially precise the activation is.   Figure 2: A bacteriograph of Michelangelo's "The Creation...

... lines (hybridomas) that secrete antibodies in culture and provide renewable reagents with reliable and...plasmid-based and offer numerous advantages, including more secure archiving, unambiguous molecular definition, and...ready-to-use-proteins to facilitate use. Dr. Trimmer’s lab will ensure the quality of the antibodies using quantitative...Reproducibility: Standardize antibodies used in research. Nature 518:27–29 . https://doi.org/10.1038/518027a Bradbury... (2015) Antibodies: validate recombinants once. Nature 520:295–295 . https://doi.org/10.1038/520295b Bradbury...

...introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! Image credit...more. This could be by outright stating the big-picture problem, by asking your audience a simple question...’t need to be anything fancy; it just needs to capture your audience’s attention enough to make them decide... new questions are next or how this will impact future research or policy.  It is worth mentioning that...patients or other affected people, man/animal vs nature, anthropomorphized animals, or even seemingly unrelated...

...CPP-tagged protein and an untagged control could enter cultured cells and activate expression of a luciferase ...After iTOP treatment, both primary (hESCs) and cultured cells showed up to 70% resistance to diphtheria...enter the nucleus and cleave the specified target. Figure adapted from D’Astolfo et al. In certain instances...elsewhere in the genome. Second, expression is by nature transient, which may lower nuclease toxicity or...Techniques: Introduction of macromolecules into cultured mammalian cells by osmotic lysis of pinocytic ...

...abilities, or other phenotypes that can be efficiently measured at the single cell level), are separated by, for...use ~400X more cells than gRNAs in a screen, to ensure that each gRNA has a good chance to contribute ...can be used: Design a screen with an easily measureable cellular phenotype (e.g. survival of a cancer...and minimize off-target effects of CRISPR-Cas9." Nature biotechnology 34.2 (2016): 184-191. PubMed PMID...and genome-wide libraries for CRISPR screening." Nature methods 11.8 (2014): 783-784. PubMed PMID: 25075903... parameters via a library-on-library approach." Nature methods 12.9 (2015): 823-826. PubMed PMID: 26167643...

... susceptibility of biofilms formed on various structured surfaces. Plasmids expressing fluorescent proteins...deletions on biofilm formation and growth will then be measured to link the deleted genes to specific functions...several diseases. Her goal is to identify and isolate pure plant compounds with antidiabetic properties that...river, take water samples at specific locations, measure fluorescent signals from the sensor, and send a.... Subscribe to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees...

...research group in Scotland, who had just the right infrastructure and offered to collaborate with them. The partnership...cutting-edge research, is still one of the most valuable features of modern science in our view. And it was this...that prompted us to create Crowdfight.   Figure 1: Crowdfight is a community of scientists who ...collaboration and join efforts to shift scientific culture towards a more generous and rewarding way of making...receiving dozens of requests every day.    Figure 2: When a researcher needs a collaborator for a...

...through the literature, to go and talk to the people who actually have these plasmids, figure out what they...heard about RNAi at his MIT interview during a lecture given by Phil Sharp. After a few rotations, Doench... screening technology.” Doench thinks that the future of CRISPR will be in high throughput functional...resonates with me.” About Addgene’s role in the future, Doench says, Addgene is “a central resource for...

...postdoc come to an end and your next professional adventure begins. While many career pathways exist for scientists...ideas every day and decide on the best way to structure roles and responsibilities as we grew.” Initially...first hires will set the tone and atmosphere that future hires will look up to." She also recommends to ...rather than repel and hoard.” If you've already secured a faculty position, start studying up before you...process. Read Genevive Bjorn's "Ready, Set, Hire" on Nature Jobs for some quick advice and resources for hiring...

...inflammatory state by inducing immunosuppressive measures, for example the upregulation of the immunosuppressive... favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive... analysis platform focused on analysing immune measures and correlating them with tumor mutations and ...locus with CRISPR/Cas9 enhances tumour rejection." Nature 543.7643 (2017): 113-117. PubMed PMID: 28225754...novel mouse glioma model using lentiviral vectors." Nature medicine 15.1 (2009): 110-116. PubMed PMID: 19122659...

...diagram below for an example of how this works. Make sure to order 5’ phosphorylated oligos or treat with ...plus a Cpf1 expression vector.  See the graph in figure 2 for a comparison of editing frequency resulting...plasmids are generally more efficient. Pro-Tip Make sure to reverse the orientation of the crRNA’s direct... used to multiplex edit 3-4 genes in a primary culture of mouse neurons and in vivo. For these experiments...Editing by CRISPR–Cpf1 Asing a Single crRNA Array." Nature Biotechnology 35.1 (2016): 31-34. Pubmed PMID: ...

...construction of our new BL2+ laboratory and the infrastructure capabilities to produce and distribute virus...laboratories all over the world. With this reach and exposure in the community, we saw the opportunity to leverage...distribution of viral samples. The Office Team made sure we could ship internationally and be ready to provide...formation of partnerships to expand the service and figure out what we can afford to do and when. Addgene’...the repository. The Outreach Team has been making sure scientists know about the new initiative and serving...

...it’s true that adding histones and chromatin architecture to every diagram portraying some aspect of eukaryotic...when I recently attended the 2018 Chromosome Architecture and Chromosome Organization Keystone Symposium...posters I saw at the 2018 Keystone Chromosome Architecture and Chromosome Organization. These aren’t the...noncoding RNA: NEAT1 RNA is essential for the structure of paraspeckles. Clemson CM, Hutchinson JN, Sara...posts below may steer you in the right direction! CAPTURE-ing Chromatin Interactions: Using CRISPR-dCas9 ...

...antibodies in adult rat brain. Figure from Andrews, et. al, Elife. Detailed figure legend here.     The ...chains as driven by two CMV promoters (orange). Figure from Andrews, et. al., Elife  A limitation ...the mAb becomes limited, which can be a threat to future reproducibility. The Trimmer Lab found that in ...recovering RNA from hybridomas grown in tissue culture, they were also able to isolate RNA from non-recoverable...

... The first thing you should do is record the manufacturer, lot number, and expiration date somewhere you...smaller than 10uL. You can leave them at room temperature when you’re working with them, but avoid freeze-thaw...okay, Blugene!    Buffers  You know what temperature to store your antibodies at, but it also matters... to react with other chemicals and biological structures, some of which you may need in your experiment...needed to excite them - and it’s a good idea to make sure that laser is available in your machine before you...

... side chains, all of which can help determine structure and function relationships or, in more practical...non-canonical amino acid at the site of interest (figure 1). While the codon used is often one of the three...system (including media, cell type, and growth temperature) may need to be individually optimized for each...variety of ncAAs to study the relationship between structure and function…if you can get your protocol to work...focused on turning GCE tools developed in the literature into robust and universal techniques that researchers...

...information and form it into a roadmap--a plan for future features, programs, or tools to build. After organizing...my boss and coworkers, discuss current and future features being built for the analysis software program...simplest of terms, a software product manager helps ensure that the company or software group is building ...

...printed for my physical lab notebook, along with any figures generated from the analyses: Goal: Before you ...will help you locate relevant information in the future. Approach: Outline a brief overview of the approach...repeating your work or making similar mistakes in the future. Stay organized Set up a table of contents in..., leave them in their own folder. Doing so will ensure that you are in fact using the same data for all...about the edit that was temporarily used. Just be sure that the original code remains in tact! Take notes...

...you meet will review your papers or grants in the future, and don’t be surprised to know that people think...about grants and projects that might help your work Tenure can depend on your national recognition and the...see enough of them).  Before the conference Make sure you have a business card – even students and postdocs... roll, say hello. Keep your cards handy and make sure your name tag is always visible. Some people learn...LinkedIn, especially if they are not in academia. Make sure to learn how to use LinkedIn properly. Then, follow...

...There are a variety of methods that can be used to measure your fluorescent protein’s propensity to aggregate...likely in an antiparallel configuration), typical structures known as Organized Smooth Endoplasmic Reticulum...your protein of interest will be problem-free. In figure 1 we provide an example of the OSER assay with ... positive and negative controls for proper ER structure. Based on these data, we conclude that mKOk has...gathered can be shared by publishing it as (part of) figure 1, uploading it as a preprint, or documenting it...

...I was studying a synaptic protein, I wanted to measure it’s levels in its normal neuronal context. In ...noncanonical amino acid tagging (FUNCAT, Dieterich et al. Nature Neuroscience, 2010) and allows the visualization...synthesized proteins in rat hippocampal neurons." Nature neuroscience 13.7 (2010): 897-905. PubMed PMID:...protein complexes in situ by proximity ligation." Nature methods 3.12 (2006): 995-1000. PubMed PMID: 17072308... of newly synthesized target proteins in situ." Nature methods 12.5 (2015): 411-414. PubMed PMID: 25775042...

...intellectual disability after Fragile X. There is no cure for CTD, which arises due to mutations in the SLC6A8...characterized by development delays, speech delays, seizures, and other symptoms as discussed in this educational...help the lab identify therapeutic targets in the future. Congratulations Mark Mellett! Siddharth Prakash... and is characterized by autism-like symptoms, seizures, and muscle weakness. So far, there have been ...patients with GNB1 mutations, the lab hopes to capture the signalosome of different receptors upon stimulation...