...apoptosis reporter, ZipGFP, which utilized the self-assembling split GFP (GFP1-10 and GFP11) to increase fluorescence...animals. To do this they modified a different self-assembling split GFP (GFP1-9 and GFP10-11) by flipping a...rectify this the Shu lab first improved the self-assembling capability of sfCherry approximately 20 fold ...FlipGFP to study apoptosis in HeLa cells, zebrafish embryos, and Drosophila. They treated HeLa cells expressing...the spatiotemporal dynamics of apoptosis during embryo development. GFP fluorescence increased over time...

...Receptor: GPCRs are also known as seven-(pass)-transmembrane domain receptors, due to the seven alpha helices...helices that transverse the cellular membrane. The receptor recognizes the appropriate ligand to activate...proteins on the intracellular side of the cell membrane. Heterotrimeric G proteins are important molecular...kit at Addgene! TRUPATH: Bryan Roth’s lab also assembled a collection of plasmids to elucidate the role...DREADDs) (Roth, 2016). DREADDs were engineered from members of the human muscarinic receptor family and are...

...they hear in their classes,” she said, “but they remember the moments when they discover something new for...focused on an emerging scientific field, BioBuilder embraces new opportunities for scientific exploration, ... Lab is a fully equipped, state-of-the-art lab embedded in LabCentral’s community of innovators that opened... Blog Synthetic Biology and the Importance of Assembly Standards Synthetic Photobiology: Optogentics ...Posts Resources on Addgene.org Guide to SynBio Assembly Standards Find Plasmids for Creating and Controlling...

...cell divisions of the C. elegans embryo when all essential embryonic genes are individually depleted via...reminded of the extent that C. elegans researchers embrace open science and share resources and tools. That...flybase. microPublication is run and maintained by members of the worm community but there are Drosophila ... lab. SapTrap is a reagent toolkit and plasmid assembly line for the creation of CRISPR targeting vectors...

...Cultured rat hippocampal neuron expressing pan-membrane ASAP5. Solid blue circle marks the initiation ...via the Kv2.1 targeting motif as well as a pan-membrane construct without the Kv motif. Altogether, if...kinetics and high responsivity near the resting membrane potential, ASAP5 is a great new option. Find ASAP5...specifically binds to active (GTP-bound) Gα. Gα-YFP assembles into functional heterotrimers with endogenous ...optional accessory plasmids, the collection targets members of all four G protein families: Gs, Gi/o, Gq/11...

...fundamental takeaway. Writing these points will help you remember what to say and provide your audience with a clear...venue and professionalism of the conference, and remember to wear comfortable shoes, as you will likely ...your breath before diving in to the next figure. Remember your outline: Instead of getting bogged down in... the minutia or coming off as a broken record, remember the outline you created for yourself. Hit the ... Resources on the Addgene website Become of a member of the Addgene depositor community Learn about ...

...start using. Prior experimenting with open source assembly algorithms meant that we already had a clear idea... individual steps to automate plasmid sequence assembly, and possible solutions for each in the form of...quality score analysis, k-mer counting, or sequence assembly), but the order and usage of the scripts needs...the freedom to use multiple open-source sequence assembly tools at the appropriate place in our workflow...Tower to augment our pipeline and enable our team members to automate their weekly sequencing tasks. More...

...Anyone could fill out the form at any time and a member of our Business Development team would review it.... Identify your enemies In Felicia Day’s book Embrace your Weird, Day identifies nine enemies that stand...Frida Kahlo or you’re stuck drawing stick figures, remember your training as a scientist has been building...world. Penguin Publishing Group. Day, F. (2019). Embrace your weird face your fears and unleash creativity...creativity. Gallery Books. Boyes, A. (2019, November 13). 5 mental mistakes that kill your productivity. Harvard...

...pic.twitter.com/sIIoIsIFd1 — Henry Lab (@EvoEmbryo) September 24, 2018   Xenopus tropicalis (Gary Gorbsky..., mix them in a tank, and observe the beauty of embryo development in real time. I was already a biology...nucleus and put it in a fertilized egg to get edited embryos (replacing the nucleus is still very difficult)...female mosquitoes. Now they are trying to replace embryo injection (hard to do, expensive, not every lab...

...specific RNA site. The EMBO Journal, 17(17), 5201–5213. https://doi.org/10.1093/emboj/17.17.5201 del Solar...plasmid copy number. The EMBO Journal, 20(24), 7323–7332. https://doi.org/10.1093/emboj/20.24.7323 Resources...post was originally written by Jason Niehaus in December 2015 and was updated by Emily P. Bentley in June...

... place, learning something new. So how can you remember everything? Write it all down! And I mean everything...second rack in the corner freezer sounds easy to remember in the moment….but you’ll probably forget and ...going to each space in order to double-check you remember where the equipment, reagents, and samples are...to ask! Especially for those new to the bench, remember to let your bench mentor know what you don’t know...polite, appreciative, and clear about their needs. Remember, your bench mentor’s training time is time set...

... common misconception is that search committee members are experts in your field. Not true. Even if you..., it's a safe bet that at least some committee members will NOT be in your area. Therefore, documents ...Talk? It is a 60-90 min session with department members to discuss your research plans for the next 5-10...," – you should show interest in other faculty members’ research. Still a Grad Student? Get Tips on Getting... running shoes and single malt whiskies, but I remember the days of modest postdoc salaries all too well...

...and breed and they have large, easily modifiable embryos, which is ideal for observing and manipulating ...majority of studies involving axolotls focus on embryonics and biochemistry. Though limited at first, advances...humans so they are often used to study vertebrate embryology and cell development. One major resource for ...understand human disease alleles during and after embryogenesis. Addgene has over 600 plasmids that contain ... a Model Organism. News-Medical. Retrieved on September 18, 2020 from https://www.news-medical.net/life-sciences...

...By breaking the transgene into smaller pieces. Reassembly of fragmented genomes is the key to fitting large...full-expression cassette. AAV’s natural ability to reassemble fragmented genomes is the basis of several split...with both part A and part B AAVs, the fragments reassemble and produce the full-length gene. Below are overviews...Vector, Big Gene Transduction: Fragmented Genome Reassembly of Adeno-associated Virus. Molecular Therapy ...of large genes to the retina by dual AAV vectors. EMBO Mol Med 6:194–211 . https://doi.org/10.1002/emmm...

...mutagenesis, and low throughput.       Gibson Assembly: An easy-to-use technique that joins homologous...be cloned into an expression vector with Gibson assembly. While this is a comprehensive mutagenesis method...method, a drawback to using Gibson assembly is that the longer the oligo insert used, the more likely it...especially single basepair deletions. Therefore, Gibson assembly is better suited for creating small, focused scanning...in the first digest (step 1 in above diagram). Remember that the BbvCI enzymes nick the DNA strand that...

...pYBL3aLK4f — Tanne van der Wal (@TannevanderWal) December 18, 2019   Holiday door The Kravitz Lab went ....com/J5bjHQo565 — Jess Tooley 🧠 (@toodlejess) December 17, 2019   Lab material tree We often get submissions..._MCDB #christmastaco #frogsnowflake #xenopus #ASCBEMBO19 #MCDB 🎄🎅🎁🐸🐸🐸🐸 pic.twitter.com/2MPLawgNQl...2MPLawgNQl — Jennifer Landino (@jenelandino) December 3, 2019 Hope you’ve enjoyed the festivities in the ...

...success, a true testament to the commitment of SITN members. SITN and DayCon Currently in its 16th year, Science...one organize a public scientific conference? As a member of the DayCon Executive Board, I was fortunate ...interactive lunch with student presenters and board members enabled attendees to engage in small group discussions...source and available on SITN’s website. 65 audience members gave up a beautiful summer Saturday to learn about...

...time, it was possible to quickly and efficiently assemble a large number of building blocks with just two... the PLoS ONE paper that introduced the kit in December, GreenGate includes the six most frequently used...Plant Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013). 2. Carola Engler et al. "A One Pot, One...Throughput Capability." PLoS ONE. 3(11): e3647 (5 November 2008).  ...

...contains the MISTIC (membrane-integrating sequence for translation of integral-membrane protein constructs...includes a special feature for difficult-to-purify membrane proteins. pCri vector pCri-13a, designed for use...protein tag. MISTIC drags tagged proteins into the membrane and may improve expression of proteins that natively...

...wanted to be able to build assemblies through covalent bonds so that the assemblies wouldn’t fall apart, even...advantage of peptide tags for building larger protein assemblies? Howarth: Peptide tags can be very generic; you...wanted to be able to build a highly multivalent assembly connected by covalent bonds, so there were no ...

...about asking hard questions. Talk to as many lab members as possible. Try to get them in a setting out of...websites, ResearchGate and LinkedIn to find past lab members. Most scientists love to talk about their past ... lab meeting. Spend some time hanging out with members of the lab. It is a warning sign if the PI does...in Mentoring" Don’t ignore warning signs If lab members, neighbors or alumni give you information about...

...as part of performing a specific function. A transmembrane protein that is being used as a marker for flow...cell, instead of when it is attached to a cell membrane.  Picking the Right Antibody  It can get even ...capture the protein bound to anything? Is it a transmembrane protein, or is it know to translocate or fold...doing or changing in different conditions. Just remember to keep good records on which antibodies are suitable...

...following year. Geoff Schiebinger, another Quake Lab member joined for half of the trip. Preparations ahead...sure the vehicle battery didn't run down. In September we arrived in YNP just as the weather was starting...Stephanie Schmidt not only produced partially assembled genomes, but also led to the discovery of IgnaviCas9...complexity with higher throughput and rigorous assembly. There, they found a CRISPR array in the resulting...

...the widely used 293T. During vector production, assembled AAV vector particles accumulate inside the producer... are often found with cellular exosomes, small membrane vesicles or microvesicles (approx. 40 - 150 nm... diameter) that are secreted from the cellular membrane into the culture medium. AAV particles are either...found that the association of AAV particles with membrane vesicles protects them from antibody recognition...

...by incubating embryos in a MG-2I solution, or injecting adult fish with the dye. Embryos exposed to just...experiments with MG-2I-dL5** were performed with embryos and adult fish. Zebrafish are an ideal model organism...organism for these experiments because developing embryos are transparent, so it’s easy to watch development...

...The Addgene Advisory Board consists of prominent members of the bioscience and tech transfer communities...apply to graduate school at MIT. Doench excitedly remembers: “When I came to MIT to interview, part of the...process was several short talks by current faculty members and it just blew me away….  I have to come here...Find CRISPR pooled libraries Learn about other members of the Addgene Advisory Board Additional Resources...

...ideas, initiative and creative thinking. The team members will be more invested in the projects and outcomes... tasks are distributed appropriately. If team members have opportunities to develop and stretch then ...then facilitate appropriate communication between members of the team to ensure that no component of the ...management style. Recipes for delegation success Remember that there are always things that the manager ...

... hires initially based on those who notice and embrace the “odd and extreme openness” of the lab (i.e ...interests, and spends one-on-one time with the current members to share short-term goals and chat about awesome...questions for a candidate to ask lab heads, lab members and lab neighbors during the interview process....alumni taken diverse career paths? Ask other lab members how the advisor treats such inquiries. If you know...

...material (BOM) of an assembly, the feasibility of manufacturing, and subsequent assembly can be difficult ... a similar way to the biological protocol, and assembly designs based around the use of commercially available...developed techniques are the key enablers for making assemblies open-source. The sharing of digital representations...

...pseudotyped to alter infectivity. Next, the virus must assemble and have appropriate factors displayed on the ...nucleocapsid viral components needed for viral packaging, assembly, and display. Pol, the last component, contains...structural capsid protein that helps with virion assembly and particle release from late endosomes when ...viruses discussed. LXs/Iva2 are primarily capsid assembly proteins required for packaging of the virus into...

...were designed by our team member Kallol Mozumdar. Sreepadmanabh M is a member of the team from IISER Bhopal...This post was contributed by Sreepadmanabh M, a member of the iGEM team from IISER Bhopal. Following up...in extensive outreach programs to encourage and embolden the spirit of scientific innovation amongst school...

...let the team detect its co-localization with a membrane-localized mCherry or GFP protein. With this system...bind, it co-localizes with its target at the cell membrane and when the OptoNB can’t bind, it localizes to...proliferation and differentiation.  Since Ras is a membrane localized GTPase, the team fused a mCherry OptoNB...OptoNB-SOS fusion protein in cells that expressed membrane-localized mCherry. With darkness, SOS binds Ras...

...quickly string together all the words that I could remember related to this paper and/or biology in general...perspective. Most people won't notice, or won't remember and those who do are very likely to be sympathetic... diving board charades, I doubt anyone but me remembers any of the incidents above–and the dumpster noises...if it doesn’t go perfectly,” can help. (And do remember to breathe!)   Public speaking as a transferable...

...-opsin GEVIs. With an increase in the neuron’s membrane potential, Ace absorbance increases and results...biologists to use Type IIS restriction sites to easily assemble multiple DNA fragments. This system has been used... for greater reproducibility as scientists can assemble parts from different labs and experimental systems...biology tools for cyanobacteria. Figure 3: Assembly of plasmids using the Cyanogate Kit and subsequent...

...interested in TS when I was a newly independent faculty member studying sex differences in thoracic aortic disease...emphasis on oligomerization, signalling complex assembly, and localization to the nucleus,” says Dr. Terry...consist of many parts that work together on the cell membrane. Germline mutations in one of these parts, the...to Addgene. Congratulations Terry Hébert! Members of Terry Hébert's lab. Additional Resources ...

..., executive director of Future of Research. On December 1st 2016, many postdocs working more than 40 hours...recently in a paper published in F1000Research on November 2016. Due to the nature of the postdoc position...been made publicly available (below). However, on November 22nd a preliminary injunction against the FLSA...for their postdoctoral salary information as of December 1st. Salary inconsistencies across geographical...

...multimerize and spontaneously assemble into particles at the cell membrane. Mature Nanoblades then bud ...CRISPR to multiple types of targets: primary cells, embryos, and animals. Nanoblades can also carry different... where they can then fuse with the target cell membrane, and transfer their CRISPR cargo into the zygote...

... to introduce genetic modifications into mouse embryos. But once you generate a growing colony of genetically... as more mutations are introduced into a single embryo, the likelihood that a mouse will end up with the... are homozygous for all of your desired genes. Remember Punnett squares from high school biology? They...transgenic mice that can impact reproduction include embryonic lethality, a short lifespan, or infertility and...

...If you were born before 1985, you might remember going to the store and buying CDs when you wanted to...antibody and when translated into protein will assemble into a fully functional antibody. These antibodies...antibody are encoded in a plasmid. (Right) An assembled antibody protein binds an antigen. Image from ...L, Xin L, Shan B, Li M (2016) Complete De Novo Assembly of Monoclonal Antibody Sequences. Sci Rep 6: ....

...Update (November 18, 2016): Researchers from a variety of institutions recently reported their inability...NgAgo is in the spotlight and take a moment to remember that NgAgo has only been available for genome ...from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA requires incubation at 55 °C - ...conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA- and Cas9...

... them may just take some getting used to. Just remember: change is good. You can experience a real onboarding...could dread these interactions, but you could also embrace them. Talking about science to non-scientists can...often are the work. Will Arnold, one of the newer members of our Scientist team says, “a well-run meeting...that much longer. If you approach the transition embracing the opportunity for growth, you can tap into curiosity...

...hopefully this post will help you get things moving - remember you're becoming an expert in your field and are...while writing, because it helps you later on to remember where you read particular studies or experiments... For example, we summarized how frequently each member of a class of proteins was mutated, as reported... a review article and not your life’s work: so remember that done is better than perfect. Good luck! ...

...becomes embedded due to the small, hook-like shape of asbestos fibers. Over time these embedded asbestos...different ways. For example, a new drug known as pembrolizumab (Keytruda) causes T-cells to more easily recognize... and cause the destruction of cancer cells. Pembrolizumab has shown some great early success in treating...

...done by creating a split dCas9 protein that can reassemble when exposed to specific UV wavelengths or chemicals...Inspired Engineering.     Alejandro (Alex) Chavez is a member of the Addgene Scientific Advisory Board. His laboratory...Mammalian Protein Complex Structure, Function, and Membership Using Genome-Scale Fitness Screens. Cell Systems...Chem Biol 11:198–200 . https://doi.org/10.1038/nchembio.1753 Tanenbaum ME, Gilbert LA, Qi LS, Weissman...

...validating its efficacy in vitro, and using it in mouse embryos to generate gene modified mouse lines. Hopefully...mammalian cells, it’s time to use them in mouse embryos. When researchers first started using CRISPR/Cas9...It takes roughly 19 days for the injected mouse embryos to develop. Once they are born, it’s time to screen...screen them for your desired edit. Good news! Remember the primers you designed to generate your pCAG-...

...excellent choice for fusions with integral membrane proteins and membrane associated proteins (i.e. GPI-anchored...including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts, the extracellular...mapping functional domains of Drosophila hsp 70." The EMBO journal 3.13 (1984): 3087. Pubmed PMID: 6526011....

... out errors like uneven pipetting or irregular membrane transfer. Figure 1: A loading control is...directly correlate to protein abundance on your membrane, which would be directly reported by band intensity...protein measurements are taken on the same gel or membrane (depending on your approach) as your final blot...of the β-actin (B) or total protein (C) on the membrane versus the amount of protein loaded on the gel...

... the fluorophores RFP (red), YFP (yellow), and membrane-tethered CFP (cyan). Brainbow 1.1 adds OFP (orange...control of Flp recombinase. The XFPs in Flybow are membrane-tethered, making this system useful for visualization... by injecting a CMV-Brainbow construct at the embryonic stage. This technique could be used to target ...cells that randomly express either nuclear GFP, membrane CFP, cytoplasmic YFP, or cytoplasmic RFP in a ...

...introverted ), and networking (how to get people to remember you). What better way to accomplish all of these...story We are biologically programmed to listen and remember better when we hear material delivered in the ... presentation faux pas, you are more likely to remember the point.   What makes a story? A story is a ... label all graphs and data professionally). Do remember the fine points of visual aids. Here’s a great...

... charged ions to rapidly pass through the cell membrane and initiate an action potential - a very useful...and close when exposed to light and change the membrane potential. OptoXRs are modified GPCRs. When exposed...downstream signaling pathways that can change both the membrane potential and a host of other cellular functions...Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proceedings of the National Academy of...