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...Fig. 2). Individual gRNAs are designed in silico using publicly available gRNA design software and synthesized...highlight some of the reagents that have been made publicly available through Addgene.  What makes CRISPR...over previous genome editing techniques is its simplicity and versatility. CRISPR/Cas9 consists of two ...essential for a given phenotype. Image Credit: www.flickr.com/photos/72005145@N00/5600978712/ What CRISPR-based...derivatives of Cas9. Several CRISPR libraries are publicly available through Addgene, with more being added...experiment, the use of appropriate controls, multiple replicates and several cell types can strengthen your results...information on CRISPR screens check out Addgene’s 2016 publication “Practical Considerations for Using Pooled Lentiviral...

...very little data on which might work in your application. It can be quite difficult to decide on which...be searched and sorted by gene/target, name, application, pass/fail rating, sample species, antibody species... how the antibody performed in the submitted application, along with a brief description of the experiment...experiment. If that data was included in a publication, you'll find that information in the report as well... section will vary slightly depending on the application — flow cytometry, western blot, immunohistochemistry...data showing how the antibody worked in your application. The submission process is easy, with simple,... through it at your own pace. To submit, simply click "Contribute Data."    Figure 6: Contributing data...

...in “crispr” and see what comes up. Before even clicking the ‘Search’ button a new content-aware suggestion...ll search for the calcium biosensor, GCaMP, and click through to the plasmid search results: More than... your lab. Instead of starting a new search or clicking through to a different part of the website, you...papers. Maybe they started their deposit prior to publication and were able to include specific plasmid ID ...terms we can use to find materials from this publication. Searching for the title of a paper works You...filters Not every plasmid search will start from a publication. You may have an idea for a plasmid that would...filters you do not need to start a new search. Simply click the ‘X’ next to any filter to clear it. Here’s ...

...transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the early genes ...encapsidate new virions) are not expressed. Note that replication-competent AdV vectors also exist and are used...reused from Kremer and Nemerow 2015 under CC-BY license.   The promise of adenoviral vectors This brings...the final topic, which is that natural AdVs, replication-competent viruses with a recombinant genome, ...Hermiston, T. W. (2003). Multigene expression from a replicating adenovirus using native viral promoters. Mol ...does not require de novo viral gene expression: Implications for gene therapy. Proc Natl Acad Sci U S A, ...in vivo and maintain activated CD8+ T cells: implications for their use as vaccines. Blood, 110(6), 1916...

...the candidate application process, the candidate’s ability to write a grant application through a written... much more than writing. In fact, many grant applications and grant makers have moved away from long written...Now they require grant seekers to use online applications with significant character limits for the written...written portion of the grant application. They have increased their demand for stronger budgets with clear...duties can range from coordinating the grant application and submitting it, to evaluating the capacity...grants profession? The number of organizations, both public and private, that qualify for grants, the amount...grant readiness, funding opportunity research, application writing and development, project implementation...

...lentiviral plasmids created for a variety of applications including cDNA expression, shRNA-mediated knockdown... thus decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells...tissues are areas of active study (for a review click here). Alternative envelopes are powerful tools ...type or tissues; this is useful in gene therapy applications where a distinct subset of the cellular population...needs to be targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all)...all) Addgene transfer plasmids are replication-deficient meaning they can be used to create virus that is...Which genes are deleted or modified to result in replication deficiency? What percent of the vector components...

... are replication incompetent and cannot readily be passed on to subsequent cells via replication like ...intracerebral application, which is more invasive and has a much higher risk of complications. References...intracellularly spread away from the site of application and aid in visualization of the morphology of...it (i.e. where the signals come from and what implications their signaling is likely to have). Anterograde...infection to the central nervous system where it replicates, spreads, causes neurotoxicity, and is lethal...HSV) also migrates through neurons where it can replicate and by doing so spread across several synaptic...cells are labeled overtime due to the continued replication of the virus. While this can be useful, if too...

... are not ideal for every application. Disadvantages include their duplicated (i.e., redundant) genome,...mutant, which regulates a developmental pathway implicated in congenital heart defects and cancer. These...its translation either by blocking initiation or splicing. (RNAi, in contrast, engages existing gene regulatory...DNA into the genome is useful for a variety of applications. For example, specific tissues such as blood...pathways control this process. Additional zebrafish applications and conclusions Zebrafish are also popular outside...—have proven useful in a variety of research applications. Lineage tracing studies highlight how these...Contemporary zebrafish transgenesis with Tol2 and application for Cre/lox recombination experiments." Methods...

...usually yes! A good cover letter won’t hurt your application, but it can certainly help you stand out.  In...consider cover letters an important part of the application. Even within an organization, there can be variance...good cover letter!) As with any part of the job application process, it's a good idea to talk to people in... to proactively address questions about your application. These could be logistical, like briefly explaining...work on [phenomenon X], I read several of your publications on [specific topic]. I am extremely interested...include a brief summary of why you are a good applicant. If you haven't talked about it yet, it's also...experiences fit the job requirements, strengthened an application. Even so, there were examples of great hires ...

...Not only is this in direct contradiction to explicit policies set up other institutions (like University...Labor issued a Notice of Proposed Rulemaking, soliciting feedback by Sept 4th.  They proposed raising ...centrally administered, which leads to many complications, including the inability of some institutions... how much they get paid. Institutional salary policies are one indicatator of how salaries may changes... 2014, 89% of institutions had a minimum salary policy: of those, 51% of institutions set their postdoctoral...National Postdoctoral Association Institutional Policy Report 2014). However, ~30% set their minima lower...lower. Furthermore, 7% did not enforce these policies. It is unclear how many postdocs are at each of ...

...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysS...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysE...Expression vector cannot have p15A origin of replication F- ompT hsdSB(rB- mB-) gal dcm (DE3) pRARE2 (...Expression vector cannot have p15A origin of replication fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo...Expression vector cannot have p15A origin of replication F-, Φ80ΔlacM15, thi, lac-, mtl-, recA+, KmR Expression... for positive selection and a p15A origin of replication, making it incompatible with other p15A plasmids...purification protocols are designed for soluble, cystosolic protein products, but this is not always achievable...

... of Visualized Experiments (JOVE). The video publication by Stuart Orkin and Daniel Bauer's labs details... size and frequency [3]. There are several applications in which the production of deletions by paired...second use of targeted deletions is to produce monoallelic deletion clones. Cells with just a single intact... boundaries may be produced to generate clonal allelic series. Single DSBs resolved by NHEJ at coding ...for deletion screening by conventional PCR. Biallelically disrupted cells can be easily identified. Even...Even in the case of clones with monoallelic deletion, the nondeleted allele is likely to bear an indel..., primer validation, and screening by PCR for biallelic deletion clones:    Like the video? The Orkin...

...of interest in separate PCR amplication reactions for recombination in SLIC cloning to form the desired...retained after SLIC cloning. The general cloning strategy described in the associated publication can be used... a critical role in development and are often implicated in disease progression. One of the major challenges...challenges in signaling research is the inability to replicate the spatio-temporal precision with which signaling...APEX (enhanced ascorbate peroxidase) reporter from Alice Ting’s lab has enabled progress on both fronts –... distinction of neighboring cells in imaging applications and the mapping of neuronal projections to their... with Sequence and Ligation Independent Cloning (SLIC). The pCoofy series of plasmids contain a variety...

...Utilizing existing relationships as part of the job application process can dramatically increase the chances... of a resume can be crucial for a successful application. It takes time to prepare an excellent resume... Many job postings will result in hundreds of applicants the day they are posted.  If your resume is not... here for more on timely submission of a job application. 3) Read job descriptions carefully Be open minded...wish list and the hiring manager knows that no applicant is likely to meet all the criteria. Note the criteria...strength you can draw on and demonstrate in your application). Beware the myths of scientist “overqualification...bench research. I am always surprised at how many applicants ignore this in their cover letters. It is also...

...et al. 2024 under CC-BY-NC-ND 4.0 International license.  Arimura et al. used MagIC-cryo-EM to determine...chaperone NPM2, but the technique should be widely applicable to other targets. The modular system of linkers...scFvs for other targets or could be used for applications beyond cryo-EM.   Find plasmids for MagIC-Cryo-EM...right). B) Protein samples protected from AWI by helical LEAs. C-E) cryo-EM single-particle analysis of ...adapted from Abe & Lim, 2024 under CC-BY-NC-ND 4.0 license. Protection by the LEAs allowed the authors...optimizing prime editing systems for specific applications! Delivering prime editors (PEs) via engineered...be delivered in a single package. In a recent publication, the Liu lab showed that this method of delivery...

... unnatural head groups for applications like fluorescent labeling via click chemistry.    Figure 4: SuperPLD...adapted from Meiring et al., 2022, under CC-BY 4.0 license. A naturally occurring microtubule-severing protein...adapted from Mahlandt et al., 2023, under CC-BY 4.0 license.   Mahlandt et al. used these Opto-RhoGEFs to study... binding of the plasma membrane. In previous publications, they described tools to control formation of...adapted from Aljiboury et al., 2023, under CC-BY 4.0 license.   Building off their earlier work on Rab11, the... et al. 2022, bioRxiv, under a CC-BY-NC-ND 4.0 license.   Just make sure you don’t overdo it! The moderately-enhanced...constructs are great for in vitro biocatalytic applications where toxicity doesn’t matter. Plus, for folks...

...Commons 3.0 license, which requires users to give appropriate credit, provide a link to the license, and indicate... have access to through your university or your public library. If you want to use pre-drawn images in...without the watermark, which are better suited for publications, if you sign up for a paid account.  I’m proud...easier to give attribution, than for journal publications. Google Image Search Google image search is ...images. For example, by selecting “Tools” and then clicking the “Usage rights” dropdown menu, you can search...always a good idea to double-check the image’s license information before using it. Have a tip for using...

...on this topic, the authors state, “Both explicit and implicit aspects of today’s postdoctoral training..., management consulting, science communication, policy and the diverse options in nonprofit science. No...science is much like industry but the “product” is publications—the pressure to produce is the same You get ...and teach students Collaborations may be less “complicated” - for example, easy to initiate a collaboration...is different about non-academia? Scientific publications are a nice to have, not a need to have Must ...every 3-6 years—a short and exciting ride Complicated matrix structures are common Small integrated...

... This allows them to replicate viral plasmids with the SV40 origin of replication. To generate a virus...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first...cell biology, there are thankfully many other applications that they are useful for! What the HEK is it...cell line and we will highlight some of those applications here.   Virus production HEKs are the go-to ...proteins at high levels. In a case where clinical applications of recombinant proteins are necessary, HEKs ...significantly reduces the cost of production. Research applications There are two areas of research where HEK cells...

...consistent, and controllable pulses of light. Complicated patterns of light intensity and wavelength can...; only two plasmids are required per system. Applications of these systems The large dynamic range of ...suitable for various scientific or engineering applications.   References Refactoring and optimization ...Oct 11. PubMed. Find the plasmids from this publication at Addgene. Synthetic biology: engineering Escherichia...:441-2. PubMed. Find the plasmids from this publication at Addgene. A synthetic genetic edge detection....2009.04.048. PubMed. Find the plasmids from this publication at Addgene. Multichromatic control of gene expression...Oct 28. PubMed. Find the plasmids from this publication at Addgene. More Resources at Addgene: Check...