...of the Cas9 nuclease (PDB ID: 4OO8 and 4UN3), Slaymaker et al. hypothesized that Cas9 cutting efficiency...enough to keep off-target sites from being cut. Slaymaker et al reasoned that, if they decreased the positive...the electropositivity of the HNH/RuvC groove, Slaymaker et al. made a variety of alanine substitutions...off-target effects genome-wide. Further work by Slaymaker et al. showed that these mutants are not toxic...SpCas9-HF1, and HypaCas9 should enable researchers to make precise edits in mammalian cells and may decrease...look forward to seeing how the research community makes use of these tools and are excited to see continued...improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally engineered Cas9 nucleases...

...technology and synthetic biology, the tools he’s making available through Addgene, and where all of it ...CRISPR is so orthogonal and programmable that you can make a very large number of regulators. That basically...have the solution. Our paper shows we can easily make orthogonal variation. The first one worked in each...off-target interactions. That’s relevant if you want to make sure you don’t target anywhere else, but for synthetic... been putting together very large toolboxes and making them available. We’ll also be submitting parts ...you want. The last few years, we’ve really been making tons of parts and characterizing the software to...sophisticated as those that living cells and animals make. That will be an outcome of genetic engineering....

...few things you can do to make the experience less stressful. First, you can make arrangements with those...company’s area of expertise. This has potential to make your audience really sit up and pay attention. Your...demonstrate technical and transferable skills that make you qualified for the position. Highlight the outcomes...improving a process or procedure, solving a problem, or making a key decision. Even deciding to stop pursuing ...4.5 hours, which was a lot of extraversion for me. Make sure to get enough sleep the night before. Have ... look professional  First impressions matter, so make sure yours counts. Dress professionally but still...

..., need to stay active and involved if we wish to make positive change. Below I present perspectives on...science happens when scientists have 1) the courage to make mistakes and 2) the integrity to recognize these...scientific evidence. We need representatives and policymakers who not only believe in the separation of church...-two-new-sciences). This is the approach policy makers take in abusing climate science skepticism to promote...averting a potential climate disaster. However, policy makers use these two fingers of skepticism to defend an...world. Justin Sherman is a Scientific Instrument Maker at Intellectual Ventures Laboratory. Vancouver, ...ongoing and difficult job. I urge other marchers to make this their first step, and continue this work by...

...tips on how to make a successful event at your company. The point of this post isn’t to make you all jealous...are busy with many other work responsibilities, we make team building activities a priority and have organized...though we are hiring!) but to encourage everyone to make an effort to build relationships with their colleagues... it simple This may be an obvious one, but don’t make things too complicated for yourself (especially ...for help My grandma used the old adage “many hands make light work” all the time. She was a wise lady for...will save you time and sanity!    10. Communicate Make sure people know that there is something fun happening...

...Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox DBT, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov...://doi.org/10.1261/rna.054098.115 Shmakov S, Abudayyeh OO, Makarova KS, Wolf YI, Gootenberg JS, Semenova...Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J,...

.... Scientists can make a new line of flies in only 6 weeks compared to months to make a transgenic mouse...in the lab, and have short generation times that make it easy to study the effects of genetic manipulations... be easily manipulated with tools like CRISPR to make transgenic lines. The fruit fly (Drosophila melanogaster...genetic material (Jennings, 2011).   The fruit fly makes a great model organism because they are easy and...C. elegans are also self-fertile hermaphrodites making it easy to conduct large experiments over multiple...60% of human genes have an ortholog in C. elegans making it ideal to study basic molecular biology processes...

...between different projects, you might not be able to make any reasonable progress on any one of them. That...another. Monitoring task progress. For this, I always make sure I have explicit goals in mind for the endpoints...everything in a Word or google doc that I can comment and make notes on as I progress. I’ve also recently started...world, procrastination can easily prevent you from making progress on a task. For instance, even if I had...over the activation energy barrier that comes with making ANY progress, the task will likely be much easier...easier. Having a formal plan for your day will make this easier - if you know you have dinner plans at 6...pm. Hopefully you’ll find this post useful for making your days, experiments, and projects more manageable...

... grow quickly and make more of the plasmids as they grow, scientists can easily make large amounts of ...guidance to understand the specific components that make up a plasmid and why each is important. Our “Plasmids...ability of plasmids to self-replicate within a cell make them attractive tools for the life scientist or ...particular environment Produce enzymes that will make specific, controlled changes to an organism’s genome...contains multiple protocols and troubleshooting tips to make plasmid usage as simple and straightforward as possible...

...arsenal to make sure people pay attention. Highlight your lowest points and highest points and make clear ...crucial part of interpersonal communication, which makes honing your ability to tell stories a similarly .... Whatever you’re working on, start wherever it makes sense to start and try to be as chronological as...protagonist In most cases, the protagonist will be you. Make sure to highlight what lessons you’ve learned and...failures. Foster conversation A story is not a lecture. Make yourself open to conversation — put your work on...

...find events stressful. Video chatting makes it pretty easy to make 30 minute appointments with people all...almost 6 months, you may be looking for more ways to make sure you are moving forward. Here are a few out-...hand at a video presentation? Most video platforms make it easy to record yourself. You can watch yourself...informational interviews, for more details on how to make the most of these exploratory conversations. If ...least two informational interviews a month you will make good progress on both building relationships and...

...and construction steps are complete, it’s time to make virus! The constructed viral vector containing the...particles are secreted into the cellular media, making viral collection fairly easy. Producer cell media...similar outcomes, so why and when should you opt to make a virus instead of just zapping your cells? That...transfect’ cells lines are amenable to transduction, making it a logical choice for efficiency. Where viruses... rarely done, viral pseudotype can be altered to make the virus specific for a type or subtype of cells...with alternative methods. Ready, set, transduce! Making virus can sound daunting, but it is easier than...

... are sensitive to blue light. Many have tried to make green molecules, but then alongside blue they actually.... So we developed a red molecule. We also had to make the blue light molecule much, much, much more light-sensitive... optogenetic tools in a deep part of the brain, making it sensitive to light. When they illuminated this... emotions, to see how a monkey changes decision making, to cure a human patient. We can't do it all by... I would say we still would like to find ways to make these tools more selective for different ions. Right...interesting. It would also be great if you could make neural silencers that require less power. Those ...

... ability to make discoveries." If you ever run out of ready-to-use virus and want to make new lentiviral...genome editing tool is its developers’ willingness to make their CRISPR technologies available to the academic... reduced titer associated with large inserts can make lentivirus production frustrating - a researcher...protocol only to find that the low titer of her prep makes it unuseable. With the large size of the SpCas9 ...biology you're interested in studying. You need to make sure your assay is scalable and robust. What you...

...components. Making proteins outside of a cell - cell free extracts Typically, if you want to make a protein...more and more molecular biology-based activities making their way into high school classrooms, like gel...cost-effective, as each one is as cheap as 10 cents, making activities designed with FD-CF reactions affordable...they can touch. With BioBits, we want students to make the connection between the facts they’ve learned...experiences and knowledge with each other to help make learning biology more fun – bit by bit. Ally Huang...

...over-diluting, and make sure to remeasure your titer after diluting to confirm. Luke Makana Hanley is a Research...Check out this post for a refresher on AAV titers Making contact with a cell Although an individual AAV ...see the dwindling proportion of AAV particles that make it through each part of the cell on their way to...additional best practices. Keep your cells happy - Make sure your cells are as healthy as can be to prepare...filter - When using a concentration filter column, make sure to rinse off the surface of the filter membrane...

...can revisit the Q&A in the chat box. Presenting? Make it interactive! As a presenter, it can be difficult...giving a research talk, there are still other ways to make your session more engaging, for example, by using...to keep your video on and look into the camera to make it seem more like you’re connecting with the people...over. Depending on the conference, organizers may make the poster available after the conference is over...getting to know the app or virtual platform and making sure you stay connected. If possible, test out ...experience as an on-site conference, you really have to make space for it,” says Bachle. “On the flip side, you...

...techniques. Show up on time (don’t make your bench mentor wait on you!), make sure you are prepared (read any...success prior to your departure for the new placement. Make sure you read any relevant literature for the techniques...you have one) to communicate your plan as well. Making logistical and funding arrangements Visiting scientist... a structured visiting program, you will have to make arrangements on your own. This will include, but...a guest and likely a trainee in your new lab, so make a good impression! Remember that your host is going...

...important part of being a “good scientific citizen” and making sure you do what you can to help your fellow researchers...consistent way so other researchers can use them, to making sure that methods and protocols are available and... this communication and sharing fits together to make for a transparent and collaborative research environment...for being transparent in their communication or making their data and reagents readily available. What...confident that everyone’s enthusiasm for actually making it happen would increase dramatically. I’m not ...resources on the Addgene blog Find more ways to make your research more reproducible Read all of our ...

... 2015. What makes lentiCRISPR v2 great? It produces a ~10 fold higher titer virus, making it ideal for...a point mutation in the puromycin cassette that makes it less efficient in some cell lines. pSpCas9(BB...V2.0 was made available through Addgene in 2015, making this the most recently available plasmid in the...a point mutation in the puromycin cassette that makes it less efficient in some cell lines so it may be... thank all of our depositors and requestors for making their reagents available and for supporting scientific...