...their research leading up to the discovery of the mechanism of dual-RNA-guided DNA cleavage in adaptive ...crRNA could be combined into one RNA to direct cleavage the result was a simpler 1 protein - 1 RNA system...cells but this can’t be used in people until we learn more about safety and more about the how to use ...cool biology can be and how much we have yet to learn. As Dr. Mello said, “Cells have been in the information...sophisticated”.     New to CRISPR/Cas9 Technology? Learn More: Addgene's CRISPR/Cas9 Guide History of CRISPR...

...England Biolabs and Labconscious. I was eager to learn about practices I could bring both to Addgene, and...impact as scientists. Many scientists are already leading the cultural change toward responsibility in our...polystyrene will likely end up in a landfill, but at least we can reduce the rate at which that happens and... it offsets the cost of regular waste services. Learn about how to start a glove recycling program at ...on culture. Encourage your workplace to become a leader in sustainability. Sign up for blog posts and...companies, and universities to be sustainability leaders. Take initiative to bring ideas to your workplace...

...exploration. Here, we delve into the importance of leaving room for serendipity during your PhD studies and...often happen when we allow ourselves moments of release and mental freedom. Practical approaches Engaging...not just a stroke of luck; it's a mindset that can lead to unexpected discoveries and innovative solutions...discoveries. So, on your academic journey, remember to leave a little space for the unexpected — it might just...Academia: Stories and Strategies for PhDs Making the Leap to Industry." He successfully transitioned to industry...Experimenting with New Careers While in Grad School How to Lead a Great Meeting Form Your Own Peer Mentoring Group...

... sites in a promoter lead to higher transcriptional activation. Multiple nuclear localization signals ...a higher signal-to-noise ratio with a lower photobleaching rate. Given the power of SunTag, they attempted... ratio Less chance of phototoxicity Reduced photobleaching Simplified, long-term single-molecule tracking...SunTag plasmids have coveted “blue flames,” it’s clear that the system is broadly applicable across many...eBook Avoid Fluorescent Protein Oligomerization Learn about Small Molecule Fluorphores Resources on Addgene.org...

...Flame Awards. What's in a flame?     We’re always pleased when scientists around the world choose to share... serves to congratulate researchers who have at least one plasmid deposited at Addgene that has been distributed...Sainsbury Laboratory in Norwich, UK: “We’re very pleased to distribute our plasmids via Addgene. Given the... us hours of work responding to requests from colleagues.” In addition to helping labs save time, we’re... plasmids and have not yet received your award, please contact [email protected] to claim your Blue Flame...deposit. Begin yours here. It has been my genuine pleasure to send these small tokens of thanks for the incredible...

...serve them better. Here’s a little bit of what we learned. Check Out Our Microbiology Page! The start of ...improving lives worldwide and the presentation made it clear how important it is for us to improve our resources...science education and outreach ourselves, we can a learn a lot from these ASM pros and hope to leverage their...was also a great place to catch up with former colleagues! I ran into past ASM President Tim Donohue (see...their materials! Resources on the Addgene Blog Learn How You Can Use Addgene's Microbiology Pages Read...CRISPR Methods for Bacterial Genome Engineering Learn how to Network at Scientific Conferences Resources...

...2017). Gam binds the free ends of DSBs, which may lead to cell death rather than NHEJ repair, thus removing...they are expressed well. The Liu lab modified the nuclear localization signals and codon usage BE4 to create...also made an adenine base editor with improved nuclear localization and expression. This base editor was... There are also RNA base editors, which you can learn more about in this blog post: CRISPR 101: RNA Editing...al., 2018 BE4max, ArcBe4max, ABEmax   Improved nuclear localization and expression Gaudelli et al., ...editing of A•T to G•C in genomic DNA without DNA cleavage. Nature, 551(7681), 464–471. https://doi.org/10.1038...base in genomic DNA without double-stranded DNA cleavage. Nature, 533(7603), 420–424. https://doi.org/10.1038...

...of your colleagues. The young scientists that are reading your peer-review comments will learn from them...delivering constructive critique can help your colleagues at the receiving end, if not for this specific...repositories will ensure that the results of your colleague’s hard work will be maintained and made available...comment differently if I would be speaking to a colleague in the office next door. So be kind, helpful, ...writing a review article Resources on Addgene.org Learn about depositing plasmids pre-publication Find molecular...

...of scientists from MIT tapped into this data to learn about trends in synthetic biology and DNA synthesis...database. Through a series of simulations and machine learning, they created a classifier that predicts whether...gene and a synthetic version of that gene was a clear predictor of whether the gene is synthetic or natural...applications in monitoring the accidental or intentional release of engineered organisms. “Is [the organism] engineered...Nielsen, Alec AK, and Christopher A. Voigt. "Deep learning to predict the lab-of-origin of engineered DNA...Addgene blog Read blog posts about synthetic biology Learn more about codon usage biases Resources on Addgene.org...

...on Precision Genome Editing with Programmable Nucleases. Check out #KSgenome on Twitter if you missed ...the CRISPR field forward. To do that, we need to learn much more about cellular DNA repair pathways. An...An emphasis on cellular repair post-CRISPR cleavage When I first looked at the conference agenda, I was...would open not with an introduction to CRISPR nucleases, but with a session on DNA repair. What stuck ...) Delivery The RNP delivery method consistently leads to high on-target editing and low off-target editing... tool supports over 250 genomes, and they just released a brand new CRISPOR manual to kick off 2018! "...

...industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends why he would...in the Boston area that demonstrate that 85% of leaving postdocs pursue a career outside the traditional... like this without sufficient data? Starting to learn about the diverse options and how to take advantage...there. Additional Resources on the Addgene Blog Learn about Possible Science Careers How to Negotiate ...for Scientists Series Resources at Addgene.org Learn About Careers at Addgene ...

...out-of-the-box ideas that might jump start your learning. 1. Create a peer mentoring group We don’t all...matter why, now is a great time to find a few colleagues to work together on career and life goals. For...scientist for show wrestling, founded a nonprofit, learned to play the ukelele...perhaps you get the idea....hope you all know by now that people get jobs at least 85% of the time because of relationships. It’s much...conversations. If you set yourself a goal of doing at least two informational interviews a month you will make...academia and not in academia is run by grant funding. Learning to write grants quickly, efficiently and effectively...

...start – it is the discussion after that will lead to learning. Listen to our podcast interview with Harvard... take informal minutes on what was covered and learned, especially noting action items that are identified...Harvard Medical School researcher Connie Cepko to learn about her mentoring style. Set goals with a mentor.... There are many advantages to being a Mentor: Learn other perspectives Develop coaching skills Encourage...Encourage your creative thinking Enhance leadership skills Support inclusion, diversity, and open communication...

...back: David Gordon, Jeff Guo, Fengbo Zhou, Paddy O’Leary (UCSF). Our approach is to discover key human...FDA-approved). This is a big job, but under the leadership of Nevan Krogan at UCSF we’ve brought together...preclinical studies already. We are following up on these leads as we speak, with collaborators in Paris and New...affected in various ways by the pandemic, it is clear that we have to stick together (6 feet apart), to...shout out to Margaret Soucheray, Jeff Guo, Paddy O’Leary, Fengbo Zhou, Helene Foussard, and Dave Gordon)... plasmids for coronavirus or COVID-19 research, please email Addgene at [email protected] with your order...

...drug discovery, and synthetic biology. Read on to learn more about the five awardees and their promising...their use as food, infusions of L. taraxacifolia leaves are widely used to treat several diseases. Her ..., the most common type of stroke and the third leading cause of death and disability worldwide. An ischemic...ischemic stroke occurs when arteries leading to the brain become blocked, resulting in reduced blood flow...awards. Additional Resources on the Addgene Blog Learn More About the Addgene-Seeding Labs Plasmid Grant...Expression Systems Synthetic Biology Resources Learn about Global Plasmid Distribution ...

...game that would force my students to use concepts learned from previous lectures, articles and books. A game...board with them. My impression was that they had clearer ideas about gene therapy in the following lessons...perhaps the ideas spurred by this “gamification” will lead to new cures for disease someday! Accessing and ...modify BUILD YOUR PLASMID or generate games like it, please freely share your projects with us! This exchange...want to receive our modifiable powerpoint file, please feel free to contact us at: [email protected] tools for teaching developmental biology Learn how BioBuilder can help teach students about Synthetic...

...while the alcohol used in the staining process bleached another, Klebsiella pneumoniae. After realizing...safranin, as a counterstain to help visualize the bleached bacteria. The chemistry behind Gram’s crystal ...drop by drop onto the smear until the alcohol runs clear and immediately rinse. Flood the smear with the ...Rinse the slide in tap water until the runoff is clear, blot gently to dry, and then view under an oil ...peptidoglycan membrane is unable to retain the stain, leading to its decolorization. In contrast, counterstain...visualizing differentiations between bacteria, leading to safer medicine and more successful experiments...

...Avoid star activity. Some endonucleases (for example BamHI) are capable of cleaving sequences which are similar...cloned the plasmid yourself or obtained it from a colleague down the hall, it is always a good idea to take...incubating your DNA with restriction enzymes which cleave the DNA molecules at specific sites and 2) running...manufacturers make High Fidelity versions of the endonucleases and/or supply custom buffers as means to avoid...is not just necessary but also an easy process. Please vist Addgene's resource for plasmid verification...

...which cleaves the synaptic protein VAMP2, a protein essential for synaptic transmission. Cleavage of VAMP2...ever like to write about a recent plasmid deposit please sign up here.      Listen to this episode of the... it inactive, thus inhibiting neurotransmitter release. How is this light-mediated? The researchers split...split botulinum neurotoxin halves to enable VAMP2 cleavage, and suppresses neuronal transmission within minutes...differentiation. Furthermore, TALEN and ZFN require custom nucleases for each gene of interest. CRISPR-Cas9 tools have...

... or profession, you need to make sure you have learned and mastered the fundamentals.   What are your ...of the office? Katz: I have a puppy and so I am learning how to train a dog with my daughter. It’s super...with our new dog, Daisy, or out on golf course! Learn more about Karen Katz, Esq Ms. Katz is the Director...Boards of BioBuilder and Brookline Interactive. Learn more about Carl Paratore, MBA, CPA Mr. Paratore ...Auditor at Tufts Health Plan. In that capacity he leads the Internal Audit function and is responsible for... MA. Additional resources on the Addgene blog Learn more about about Addgene in our "Inside Addgene"...

...specific. Once inside the host cell, the virus unleashes its cargo – its own genome.   Fig. 1:...access to the host genome requires breakdown of the nuclear envelope, which occurs during mitosis. The most...and dividing cells. They accomplish this through nuclear localization signals which can traffic their viral...PMCID: PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human immunodeficiency virus type 1 preintegration...PMC1368960. Additional Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss...Resources on Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral ...

... two results for your antibody of interest: two clear populations or a huge smear across your FSH vs reporter...intensity. Signals any higher than this may not be clearly picked up by the sensors. You can adjust this by... positive population between 103.5–104.5 for a cleaner readout. A yes-no plot If you are using an antibody...sample (right). Negative and positive samples are clearly distinguishable as two separate populations. Created... data as a dot plot.  To summarize, if you have clear positive and negative populations for your marker...compare between samples. In cases where there is no clear distinction between the presence/absence of your...

... see by this post’s chromatin illustrations from Leah Bury of Microscopic Art. I started thinking about... we definitely don’t have that you really want, please let us know! And if you study chromatin and want...indicated gene, and you can subscribe to these pages to learn when Addgene receives new plasmids containing these...t see the gene you’re looking for on this list, please consider searching directly on our website. ... 25. 95(5):625-36. An architectural role for a nuclear noncoding RNA: NEAT1 RNA is essential for the structure...Addgene blog post about chromatin--multiple Addgene colleagues and guest bloggers have written posts describing...

...affected by light. Long-term light exposure can also lead to cellular phototoxicity, and experimental success...can be affected by both autofluorescence and photobleaching. Researchers have long been interested in using...reality, Addgene depositor Takeharu Nagai and colleagues at Osaka University have developed Nano-lantern... cells. As Nano-lanterns are not subject to photobleaching/phototoxicity, they successfully imaged multiple...applications. For long-term imaging, especially where photobleaching and phototoxicity are a concern, these constructs...Addgene. Additional Resources on the Addgene Blog Learn about Photoactivatable Fluorescent Proteins Check...

...Blog Learn About our Cancer Colleciton Pages Use the ClonTracer Library for Lineage Tracing Learn About...to well-annotated cellular pathways). It wasn’t clear whether gene-agnostic bioassays could accurately...proud to have worked closely with our Addgene colleagues to make these available to the scientific community...10.1016/j.coisb.2017.02.002. We hope that by releasing these new and exciting plasmid collections (together...clinical sequencing studies. At the Broad, my colleague Cory Johannessen and his team have recently demonstrated...

...polymerases (Pol μ and Pol λ), nucleases (Artemis), and structure-specific end cleaning enzymes (Aprataxin, Tdp2...Cas9 protein doesn’t immediately release from the broken end after cleavage, which may interfere with loading... the products of accurate repair are easily re-cleaved while indel products aren’t, so repeated cycles...latter products. As noted above, a single cycle of cleavage and accurate repair should take less than an hour...

...activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or genes to...published in Nature in December, Konermann and her colleagues in Feng Zhang’s lab describe just such a system...transcription activation processes. Konermann and her colleagues demonstrated the new system by screening for ...in those 15,977 human genes. Weissman and his colleagues demonstrated in a paper in Cell that they could...which guide RNA we use. What we have to do now is learn more about the rules.” He said that there is no ... These libraries are lentiviral in nature, so please ensure that you are equipped and authorized to ...

...review see Maday et al., 2014). It is currently unclear which proteins facilitate transport of neuronal...expansion of RABV. This improvement resulted in a clearer picture of neuronal connectivity in the mammalian... mapping of the neuronal circuitry involved in learning and memory (Kohara et al., 2014; plasmids at Addgene...particle is taken up on one side of the cell and released again at another side of the cell after intracellular...transport without actually causing infection or leading to transgene expression within the first cell. ...development of more powerful AAV serotypes, and perhaps lead to controlled trans-synaptic spread of AAV vectors...21782903. McFarland, N.R., Lee, J.S., Hyman, B.T., and McLean, P.J. (2009). Comparison of transduction efficiency...

... and ligands vary in their chemical structure, leading to differences in their excitation and emission...peak wavelength can also decrease the rate of photobleaching. Interaction with receptors: Understanding ...the choice of fluorescent dye. For instance, the cleavage of FITC-conjugated protein ligands is unaffected...experimental conditions. Common issues include photobleaching, non-specific binding, and quenching, which... reduce ‘muddying’ of the signal and facilitate clear differentiation between multiple dyes during analysis...states in response to a specific stimuli, which leaves only a small volume of fluorophores in an excited..., M., & Ghods, R. (n.d.). Comparison of the Photobleaching and Photostability Traits of Alexa Fluor 568...

...that other technologies need to be discarded but clearly we are in an era of neuroscience research where... the technique is remarkably straightforward to learn and relatively cheap (compared to many other technologies...Program (NIH-NIDA IRP), with the stated goal of learning behavioral optogenetics in order to implement ... to share some of the tips and tricks and I’ve learned from numerous scientists at NIDA and some of the...region can result in a particular neurotransmitter release or post-synaptic current. Fortunately, if your ...variants [1, 2, 4]. And, of course, the best way to learn about the many different opsin variants and how ...University.  His interests include the role of learning and memory in opioid addiction and science communication...