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...toolbox. However, both of these proteins are quite large — comprising approximately 1000–1400 amino acids...challenge for efficient delivery into cells, particularly via viral vectors. While viral delivery is a... cell types, it has a limited capacity to carry large DNA cargos. As a result, the search for smaller,... Cas12a, uses just one to cleave both strands. Similar to how Cas proteins require a PAM (protospacer ...capacity, which makes it difficult to accommodate the large coding sequence of commonly used nucleases like ..., Nasti, R. A., Vollbrecht, M., Starker, C. G., Clark, M. D., & Voytas, D. F. (2019). Plant gene editing...an Old Favorite A History of Genome Editing in Popular Culture Additional Resources on addgene.org Genome...

...contributed by Kutubuddin Molla, a Fulbright Visiting Scholar at the Pennsylvania State University. Imagine you...Later the resultant U-G pair is either repaired by cellular mismatch repair machinery making an original C-G...’ to ‘I’ (inosine), which is treated as ‘G’ by cellular polymerases. Subsequently, an original genomic... is converted to a G-C pair (Figure 1). As the cellular inosine excision repair is not as active as uracil...Figure 2: Plant base editor platforms and their molecular components. Adenine base editors for plant.... Currently Kutubuddin is a Fulbright Visiting Scholar at the Pennsylvania State University, University...· T to G· C base editing in the rice genome." Molecular plant 11.4 (2018): 627-630. PubMed PMID: 29476916...

...publications Online tools like ResearchGate, GoogleScholar, Academia, ORCID, and MyScienceWork all help... especially published research, have grown in popularity over the last decade. Research by Scott Spicer...abstracts identified. The journal Cell had the largest average view count per video abstract at 2,947 ...being used to share science. TedTalks are very popular, though they focus mostly on highly innovative ...even use a computer.  Currently, two of the more popular biology videos are:     “Singing” Fish Rely ...Ryan at [email protected]   References 1. Larivière, Vincent, Yves Gingras, and Éric Archambault. ...and case study." Journal of Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources...

...scientific or scholarly research contributions in the field Evidence of authorship of scholarly books or articles...should not be relied upon for legal advice in any particular circumstance or fact situation.  The information...agencies to propose reforms to the H-1B visa system. Large firms and firms that work on projects with an international... extraordinary ability category The EB-1A - is similar to national interest waiver criteria however the...showcases (not usually applicable to scientists). High Salary –not relevant to most postdocs. Commercial successes... articles (in scholarly journals with international circulation) in the field EB-1Bs historically have...documents that are not in English. Create a Google Scholar account Sanitize your LinkedIn, Facebook profile...

...Biology have shown similarly disappointing rates of reproducibility. These alarming results are due, at...publications and funding. If any of these three pillars crashes, the lab's success and the careers of its...effective, and our schedules more organized. From the alarm in the morning, to the daily news briefing, to quick...solution that fits the most labs, our team conducted a large study which focused on the adoption of ELNs in labs...talking about ELN costs and limited budgets, “a large percentage of survey respondents indicated that ...support that. In addition, if you are working for a large institution or company, site licenses might be available...

...to efficiently deliver large repair templates for applications that require large gene fusions. The current...vehicles for episomal gene expression and are particularly useful for gene delivery to the nervous system...is composed of single stranded DNA, which is particularly well suited for strand invasion, a key step ...edit your gene of choice. These vectors are very similar, both with polylinkers for the addition of gene-specific...gene targeting in human pluripotent stem cells." Molecular Therapy20.2 (2012): 329-338. PubMed PMID: 22108859...Matthew H. Porteus. "CRISPR-mediated integration of large gene cassettes using AAV donor vectors." Cell reports...

...plasmids together on a tight budget. To help popularize our kit resources,  we’re highlighting just a...seven-repeat proteins: Sjuts, Hanno, et al. "Molecular basis for inhibition of AcrB multidrug efflux ...open (2016): bio-017129. Hsia, Justin, et al. "Modular Synthetic Inverters from Zinc Finger Proteins and...translation initiation sequences are optimised for a particular system. The Alexandrov lab at the University ...University of Queensland’s Institute for Molecular Bioscience has provided a solution for this problem by developing...changes in enzyme function can lead to surprisingly large fitness effects during adaptive evolution of antibiotic...specific RNA bases. By swapping out these different modular repeat segments, you can create a PUF protein that...

...relatively low postdoc salaries as opposed to Staff Scientists Increase the Starting Salary for Postdocs to ...organizers wanted to touch on in organizer David Riglar’s blog post. Essentially the goal was to determine... base of all these questions as there is not particularly good data on how many post-docs there are in...positions outside of academia Require that postdoc salaries be paid on fellowships instead of grants such ...to $50,000 (the current median postdoc starting salary as reported by Paula Stephan is $40,000) Increase... Found Here Resources at Addgene  Read David Riglar's FOR Boston 2015 Blog Post Read Our Career Blog...

...personnel salary ranges, go to online job information sites (like, indeed.com, or salary.com). Search ...institute is located. For example, salaries in Kansas versus salaries in NYC will vary widely. Therefore...institution/funding source. Major equipment includes large centrifuges, sequencers and analyzers, and any ultra...candidates, and, more importantly, it’ll help you define salary costs. 4. Ascribe a cost structure associated to...Therefore, start up awards should reflect salaries in the region you’ll be working. At this point, your budget...

...Double-Strand Break Repair in Mammalian Cells. Molecular and Cellular Biology, 18(1), 93–101. https://doi.org...search for "CRISPR", short-hand for Clustered Regularly Interspaced Short Palindromic Repeats, in Pubmed...Matt Ferenc, to organize the CRISPR forum's most popular questions and answers. Designing your CRISPR genome...introduce cleavage of the target site with efficiency similar to using a single chimeric gRNA. At the same time...with the Cas9/sgRNAs vector (like PX330). For large changes (>100 bp insertions or deletions), we typically...lab typically use PAGE purified long oligos. For large changes (>100 bp insertions or deletions), the Zhang...Addgene blog CRISPR Cheat Sheet Build Your CRISPR Vocabulary CRISPR 101: Off-Target Effects Visit the CRISPR...

...possibilities enabled by this highly modular AAV - that’s probably why it’s so popular! We’re excited to see how ...serotypes AAV1, AAV5, and AAV retrograde (the most popular form), this viral vector is used to deliver DNA...specific Cre can provide you with that control in a modular way. For example, say you would like to label ...Cre-controlled gene of interest makes pmSyn-BFP-Cre highly modular. Further uses of pmSyn1-EBFP-Cre Of course there...there are also more complicated ways to use this particular AAV. For instance, you might use it as part of...

...reinventing the wheel is inefficient. An effective team regularly seeks expertise wherever it can be found. Listen...allies" – these are employees in your group that regularly step up to give feedback and fill you in on percolating...constructive input from your team: Solicit feedback regularly and often A one-time event doesn't work to get...in non-public settings. Ask over coffee.  Have regular 1:1 meetings with team members (don’t become known...experts recommend using anonymous surveys with regular frequency. Anonymity can be a great way to get ...patience, pause; A = ask at least one question to clarify the situation; C = confirm that you have understood...

...scientists want to transfer large genes, smaller binary vectors with a larger cloning capacity would be ... of DNA that will be transferred to the plant, similar to viral vectors’ terminal repeats. It’s a lot ... most first-generation binary vectors are quite large (>10 kb.) Since transformation efficiency is inversely...pKYLX vectors, respectively, making it ideal for large gene transfer. The M24 promoter provides constitutive... the plasmid can also be customized due to its modular structure. Two multiple cloning sites (MCS's) flank...

...cells infected with malaria or other parasites. Addgene: Do you have any particular tips or tricks for ...generating new tools to manipulate biology based on molecular features found in nature, with the ultimate goal...improve the diagnosis of disease, and cancer in particular. They recently introduced the SpyTag/SpyCatcher...What is the advantage of peptide tags for building larger protein assemblies? Howarth: Peptide tags can be...systems and they shouldn’t get in the way much. Particularly we wanted to be able to build assemblies through...

...over the years, he and his colleagues built a Molecular Expressions collection including some 3,300 plasmids...designed in the same vector backbone with very similar restriction enzymes. You can essentially cut and...of many proteins known to localize to specific cellular structures.The Davidson lab even included some...his team created with the research community at large, "[Fluorescent proteins are] a whole new way of ...everywhere to get a clearer picture of that intracellular world, even as the Davidson lab prepares to ...scientists, and the FSU team who have made this large, important plasmid deposit possible.   ...

...Haoyi Wang! Wenning Qin is a molecular biologist with Biogen and is particularly interested in applying CRISPR...the go-to choice for studying mammalian biology. Similar to any model, mice are not without their problems...manipulate the mouse genome using CRISPR/Cas9. Similar to the human genome, the mouse genome is made up...exciting – the cell sends out an “SOS” signal and cellular repair mechanisms rush in to repair the damage...generating small mutations with HDR, when it comes to larger scale genome editing, such as replacement of a ...degradation by the omnipresent RNase. You may want to declare your bench “RNase free” and refrain from talking...

...Transposagen. He has a long-standing interest in molecular and cellular biology and brings 10 years of research...mutations behind. Furthermore, piggyBac™ offers a large cargo-carrying capacity (over 200 kb has been demonstrated...Controlling the expression levels of your gene may be particularly useful when the gene being expressed has threshold...accessory transposon sequences.  The use of PBx is particularly important to avoid any possible random re-integration...a cell line with the edit of interest. With the larger piggyBac™ donor vector, it is possible to make ...recombinase. However, the use of negative selection largely overcomes this obstacle. Furthermore, we have found...

...Resources on Addgene.org Molecular Biology Reference Pages Basic Molecular Biology Protocols Diagnostic...feature detection with an easy to use interface. We regularly use Snapgene for our quality control process because...will confirm for us is whether it is a linear or circular sequence. Next, we use the “Detect Common Features...analyzing deposits with many plasmids that contain similar features and it allows you to more quickly identify...annotated in our plasmid maps, including our most popular primers and certain restriction enzymes, but you...

...Drawings is similar to using PowerPoint to draw figures. It’s part of Google Drive so it has a similar interface...figures quickly. Inkscape Inkscape is the most similar to Adobe Illustrator out of the three options for...like lab meeting presentations. You can download larger images without the watermark, which are better ...four main categories: anatomy and the human body, cellular biology, medical specialities, and general items... tools settings let you tailor your search to particular types of images. For example, by selecting “Tools...

...scientific sharing worldwide but a select few are particularly special. Over 1000 of our 60,000+ deposited ... the way in which we share plasmids and other molecular tools with the scientific community worldwide,...disseminates throughout the research community. These popular plasmids are categorized into a 3-level system ...can be a time-consuming chore for labs that have popular plasmids but lack the resources or personnel to...our plasmids via Addgene. Given the increasing popularity of CRISPR in the plant community, this has saved... as a whole. To all of our depositors- and in particular our Blue Flame Award winners- we’d like to thank...