We narrowed to 1,042 results for: URE

...applications.  Third, even in the most optimized tissue-culture based system, the efficiency of gene editing is...limitations, the AAV approach is best suited for tissue culture based approaches that require modification of only...mutations of a gene into the endogenous allele in cultured cells. AAV genome editing combined with targeted...T-cells and hematopoietic stem cells respectively. Future researchers could apply this technique to efficiently...Hirata. "Human gene targeting by viral vectors." Nature genetics 18.4 (1998): 325. PubMed PMID: 9537413...editing via postnatal administration of AAV vector cures haemophilia B mice." Scientific Reports 7.1 (2017...

...Scientists. Back when I used to do mammalian cell culture, I would often respond to friends asking me to ...achieving your goals. However, you should have a good mixture of useful tasks that can be accomplished quickly...Monitoring task progress. For this, I always make sure I have explicit goals in mind for the endpoints ...extensively, my lack of desire to go sit in the tissue culture hood may have prevented me from splitting cells...splitting cells boring, hanging out in the tissue culture room also gave me time to listen to some of my ...any other time management suggestions, please be sure to note them in the comments! Additional Resources...

...biomedical research and drug development. Integrin structure and conformational complexity There are 24 known...With the right research reagents, researchers can measure this conformational change to investigate the role...specific integrins in cellular processes.   Figure 1: Integrins can shape into three conformations...much lower affinity than their open states (C). Figure courtesy of IPI. IPI anti-integrin toolset IPI...biology talks on the IPI website and stay informed on future events by signing up for the IPI newsletter. The...characterization of subtype-specific β1 integrin mechanics. Nature Communications, 13(1), 7471. https://doi.org/10.1038...

...Make sure to check out our validated gRNAs to avoid rolling the dice on a new gRNA in the future! References...target cells with Cas9. Hooray! Now it’s time to make sure your genome edits went according to plan. In this...validation on your bulk population. This way you can ensure you aren’t looking for a needle in a haystack before...sgRNA-Cas9 combo. Pro tip: When designing your PCR, ensure there is at least ~200 base pairs of sequence flanking... inserted or deleted accordingly. Pro tip: make sure your PCR amplicon isn’t too large so that you can...

...to recapitulate the true steric and charge-based nature of the phosphoryl-modification; they are both much...defined three components required for translation (Figure 1): A tRNA that recognizes a specific ‘open’ codon...encoding the open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the...be optimized for parameters such as induction temperature and length of induction, which will vary between...Philip. "The origins of protein phosphorylation." Nature cell biology4.5 (2002): E127-E130. PubMed PMID:... permits programmable protein phosphorylation." Nature communications 6 (2015). PubMed PMID: 26350500....

...editing efficiency. Read the paper in Nature Find the SECURE base editors All-in-one AAV delivery ...inhibition Covalent tethering of DNA template to Cas9 SECURE base editors Nme2Cas9 CRISPR interference in Candida...genome as a negative control. Read the paper in Nature Communications Find the E. coli genome-wide CRISPR...plasmids Selective curbing of unwanted RNA editing (SECURE) base editors Keith Joung’s lab recently found ...

...perform transformations. A typical 200mL bacterial culture produces 20mL of competent cells, enough for 800...bacteria, the bacteria is typically grown in liquid culture without antibiotics to give the antibiotic resistance...plate your cells without adding plasmid) to make sure you don’t have contamination.  Also conduct a ...that transforms well in other competent cells) to ensure that your cells can be readily transformed. Compare...previous batches of homemade competent cells to ensure good efficiency.  Want even more tips for transforming...

...popular any one of them will become. One thing is for sure, as is obvious from our most requested plasmid, ...modified for viral trasduction (see the Zhang Lab's Nature Protocols Paper for more information). V2.0 This...included in this plasmid's name are some additional features. The first and most imporant is that, upstream...these variants, as well as links to the Zhang Lab's Nature Protocols Publication describing them here. References...Genome engineering using the CRISPR-Cas9 system." Nature protocols 8.11 (2013): 2281-2308. PubMed PMID: ...

...AacCas12b) exhibits optimal DNA cleavage at 48 °C, a temperature incompatible to genome editing in human cells...strand breaks at target sites. Read the paper in Nature Communications Find the Cas12b plasmids  Plant...AAV toolkit - this time using the Staphylococcus aureus Cas9 (SaCas9). This AAV-based genome editing system...highly customizable as vector backbones, and gRNA structure, numbers, and promoters can all be chosen. Read...transferred via conjugation.   Read the paper in Nature Microbiology Find the Mobile-CRISPRi plasmids...

...Based on CRISPR/Cas by Shravanti Suresh Guest blogger Shravanti Suresh who studies CRISPR at Iowa State...launched the cloning grade DNA service. This service features 210 plasmids: empty backbones, fluorescent protein...plasmid in TE, Tris, or water? And what about temperature? Find the details in this blog post. 9. Droplet...stably maintain them in the cell. This blog post features two mechanisms that cause plasmids to be incompatible...

...narrowly focused lectures, interactive panels, and a student poster session. All DayCon lecture content is ...variety of programs, including fall and spring lecture series, science cafes, and Reddit AMAs. Their website... engaged. The day kicked off with introductory lectures targeting three broad concepts: the genetic basis... what life is like as a student researcher. The future of DayCon DayCon and other similar events show ...

...really made me think about my future. They made me rethink having a structured career because our mindsets...how to translate them into real world careers. I figured this internship would be a perfect way of exploring...different experience, It was helpful to have Addgenies reassure me that someone would always be there to help ...experience at Addgene, I am excited to see what my future looks like. ...

... toolkit SECURE adenine base editor Earlier this year, Keith Joung’s lab developed SECURE cytosine base...compared to rat APOBEC1. Now, they’ve engineered SECURE adenine base editor variants using a protein-truncation...RNA-recognition capabilities.  Read the paper in Nature Biotechnology Find the base editors   Single base...editing efficiency of a target site compared to a mixture of two base editor enzymes in mammalian cells. ...

...et al., 2024).   Figure 1: Schematic showing the X-CHIME approach. Figure from LaFleur, et al. 2024...sequential knockout of genes in the immune system. Nature Immunology, 25(1), Article 1. https://doi.org/10.1038...cell subpopulations and restrains tumor immunity. Nature Immunology 20, 1335–1347. https://doi.org/10.1038...in vivo screening of genes in the immune system. Nature Communications, 10(1), 1668. https://doi.org/10.1038...

...climb upwards until they reach a point where the temperature and humidity favors fungal growth. The ants bites...out of the ant’s body. The fruiting bodies then rupture and release spores onto the forest below for the...Vampirovibrio chlorellavorus will not grown in culture alone and relies on its algae host for reproduction...through its gills. It attaches there until they mature and changes sex from male to female. Then, it detaches...resources on the Addgene blog Read about company culture at Addgene See some of our previous Halloween costume...

... on beneficial opportunities or to find oneself unsure of the next steps. Harvard PI Angela DePace has...personal development at the center of her lab (pictured above). DePace holds yearly planning meetings ...trainee to create a supportive, growth-oriented lab culture. The aim of these meetings is to give honest, constructive...PI-trainee communication builds a positive lab culture Although they may take some getting used to, yearly... lab culture...

...screens, but not to know even the life cycle of the creature they deal with.” So photography can be used as...2008. His work exposes and explores the hidden structures within flowers with the use of x-rays. His haunting...and have gathered much acclaim - including being featured in the National Geographic and Smithsonian Magazine...scientist with a new method to explore the inner structures of delicate plants and other organisms without...interest for its stunning images that is has been featured in Time magazine . Other smaller competitions ...

...nLightR (Kagiampaki et al., 2023).     Figure 1: Properties of nLightG and nLightR. A, Structural.... Scale bars, 10 μm (HEK293T), 20 μm (neurons). Figure and caption adapted from Kagiampaki et al., 2023...enhanced response time allows for more precise measurement of norepinephrine uptake throughout the brain...indicators for monitoring norepinephrine in vivo. Nature Methods, 20(9), Article 9. https://doi.org/10.1038...ColabFold: Making protein folding accessible to all. Nature Methods, 19(6), Article 6. https://doi.org/10.1038...

...It was immediately clear that Addgene wanted to ensure applicants understood what the expectations were...responsible for helping other scientists on the team ensure the deposits we received are what the depositing...for me at Addgene. There is a clear effort to make sure people feel connected.    As I became accustomed...aided by the efforts being put in by Addgene to ensure their employees are feeling comfortable.     Lucie... Tips for Writing a Good Cover Letter 7 Tips to Secure a STEAM Internship This Summer  ...

...They generated these editors using the BE4max architecture. The new editors reported in this study have... levels relative to BE4max.  Read the paper in Nature Biotechnology Find the base editor plasmids CRISPRi...CRISPRi in Caulobacter crescentus Figure 1: Serial dilutions of CRISPRi in Caulobacter cells for...Enhanced CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout... methylation-silenced genes. Read the paper in Nature Communications Find Casilio-ME plasmids If you...

...called “self-service.” The idea here is to build features that allow users to help themselves, saving time... will continually update the Help Center to make sure that, whatever your support needs are, we’ve got...to the Help Center, you may have noticed a new feature on our website; towards the bottom right corner...www.addgene.org. We’re excited to make these new features available to the Addgene community, and we’d love...Read other Inside Addgene Posts Learn about New Features on Our Plasmid Pages Resources at Addgene Visit...

...was delighted to be invited to the ceremony for surely what is one of many awards that will be bestowed...in markers on a whiteboard. The final image is pictured here…she started from a blank white slate and ...experimentation to the eventual translation to the clinic and cures. There is no doubt that companies are swarming to...computer to fight the known viruses. Bacteria have figured out how to download a bit of virus sequence to ...example, the technology is already available today to cure sickle cell anemia in stem cells but this can’t ...

...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...

...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...

...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 ...

...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...

...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 ...

...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...

...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...

...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...

...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...

... 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...

...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...

... 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...

...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...

...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...

... to increase capture. Longer histidine tags are also recommended if you need a very pure sample. Because..., alternative fusion tags were used, such as S. aureus’ Protein A (Moks 1987). These tags, however, tended...denaturant. Not all proteins will refold once denatured, however, so take care when using this method....purification of human insulin-like growth factor I from culture medium of E.coli. Bio/Technology. 1987, 5:379-382...