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...disciplines – you can select the disciplines and the years. Which leads to the question of how we can support researchers...wesharescience.org. Sharing your research with the world can be challenging. After months, or years, of grueling...uncited. Creative titles and controversial topics can garner some attention, though in reality much of ... explore new connections to our research. So how can we fault others for failing to find our work?  I ...in the article is an interactive chart where you can see the yearly interdisciplinarity index of research...short, 3 to 5 minute videos about research. Videos can be shared by research scientists, faculty, students...challenging even when it is of quality). On the site you can, for example, learn about how researchers are using...

...common activities can be done straight from the new search bar, but the menu just above it can help you quickly...make your experience on our website better so you can get the resources you need more easily.  Here’s ...if you’re in the mood to browse our catalog, you can use the links under the search box to go straight...   Figure 1: The new search bar.   Pro tip! You can use a manuscript title to search for plasmids from... our science options into a single place, so you can quickly navigate around the Addgene website without...

...- links to articles citing this plasmid. Now you can learn how a plasmid has been used by multiple labs...listing the articles that cite this plasmid. You can use the dropdown to increase the length of the list...list (purple oval in the screenshot below). You can also use the "Search Table" box at the upper right...list of citing articles. From the article list you can click on the PubMed link to find the article abstract... database - and we always appreciate any help we can get. If you know of an article that used a plasmid...plasmid from Addgene, we want to hear about it so we can link to it in the "Articles Citing This Plasmid" ...

...tissue-specific diseases, as you can target your library to a significant degree using different types of...       In vivo, in vitro, ex vivo CRISPR screens can be carried out using a few different methods — in...infected with the CRISPR library. In vivo screens can become costly, so having an efficient and effective...delivery method for in vivo CRISPR screens. AAV vectors can be made at high titers equal to those achieved with...affect which is chosen for certain experiments. AAV can be safely handled at BSL-1, while lentivirus requires...vectors being packaged using VSV-G. In contrast, AAV can be packaged in a variety of capsids. To date, at ... interest — for example, AAV8 for kidney cells — can help improve infection rates in your experiment. ...

...secondaries, can improve signal strength (Sherry and Stiles, 2022). Fourth, signal continuity can appear reduced...they can achieve. This means that regardless of how powerful of an objective you have, you can’t resolve...is limited in the number of specific targets you can label in a sample. Super-resolution microscopy (SRM...original size. Depending on the protocol, samples can be expanded to ~4-10x linearly! This expansion physically...said, the anchoring, digestion, and expansion steps can impact the integrity of different targets and labels..., et al. 2020). With any variation, your samples can tear or expand unevenly, introducing distortions ...expansion results in dimmer samples. Reduced signal can be caused by loss of protein during digestion, or...

...fluorophore can emit a limited number of photons before bleaching. The amount of information one can extract...overexpression of protein fusions to gene-edited cells can decrease the number of fluorophores, compromising...budget, which determines how long and precise we can track individual molecules. Poor photon budgets are...fluorescent proteins—even the thriftiest cell biologist can feel like a college student searching couch cushions...more photons from a sample. Chemical fluorophores can be substantially brighter and more photostable than...transcyclooctene–tetrazine)– Nonnatural amino acids can be incorporated into a protein structure and then...1e) are inherently fluorogenic and several others can be made fluorogenic using environmentally sensitive...

... of a particular observation can be easily studied as each component can be examined independently. In...thus modeling genome replication. These products can be detected by Northern blot analysis. In addition...producing firefly luciferase. Firefly luciferase can readily be measured to quantify (indirectly) transcription...by the viral polymerase. Alternatively, the mRNA can be detected directly by Northern blot analysis or...In addition, experiments can be performed in the absence of some viral proteins (compared to viral infection...Therefore, the roles/functions of individual proteins can be studied independently of other viral proteins ...simplifying the interpretation of results. This system can be used to develop an understanding of and manipulate...

...the ‘bait’) can be ‘fished out’ for further study. In Y2H screens, a group of prey fusions can either be...experiments. False positives can occur even in small scale experiments, although these can often be accounted for...cellular compartment or growth stage). A prey library can either be applied to the bait in a single population...false negatives; up to 75% of known interactions can be missed by single-pass two-hybrid screening (6)...levels of bait and prey proteins - overexpression can lead to false interactions. Lowered expression levels...for problems with one or more of these parameters can lead to a high number of false negatives in Y2H screens...responsible for the modification in the assay host strain can help. In order to reduce the chance that the configuration...

...titers can drop 10-fold for every 2kb increase in insert size. To overcome this, lentivirus can be concentrated...popular gene delivery tools, producing lentivirus, can pose certain challenges. Whether choosing a system...competent lentivirus (RCL). Replication competence can arise from recombination events during viral production...fourth plasmid. The requirement for additional DNA can impact the transfection efficiency of the packaging...Requiring an additional plasmid for transfection can make these optimization experiments a bit more challenging...transfection conditions. As mentioned above, plasmid ratios can be adjusted to enhance titer. In addition the transfection...for an example of how changing transfection ratios can affect transfection efficiency. For some transgenes...

... where one can publish brief novel findings. These results are normally one figure and can be negative...Using SapTrap, this CRISPR plasmid can be created in a single tube. You can also find additional SapTrap plasmids...and Sibernetic, a fluid mechanics simulator that can be used to simulate muscle tissue as it applies to...microPublications are peer-reviewed and assigned a DOI that can be referenced. In addition the data published in ..., 2015). In this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin dependent..., a GAL-4 based bipartite expression system that can regulate gene expression in C.elegans. In this system...promoter controls the expression of a GAL-4 driver that can bind to activation sequences upstream of genes to...

... primary cells, embryos, and animals. Nanoblades can also carry different types of CRISPR cargo including...Nanoblades then bud into the cell culture supernatant and can be purified by centrifugation. Gag-Cas9 fusion protein...the two proteins, thus releasing Cas9. Nanoblades can also deliver Cas9 variants, such as a transcriptional...Gag-Cas9 fusion protein. A single Nanoblade particle can be loaded with 1-4 different gRNAs. Viral envelope...showing that the method of Cas9 and gRNA delivery can influence the level of off-target effects, with electroporation...carries a gRNA, Cas9, and a HDR donor template DNA can be generated by incubating Nanoblade particles with...zygote which is both technically challenging and can negatively impact zygote viability. Nanoblades, however...

...remodeling can occur by two main mechanisms, either through histone modifications, which can influence ...hunger that can lead to excessive eating and obesity. What is fascinating is that both syndromes can present...on top of the genome’). Epigenetic modifications can result in phenotypic changes without any change in... have very different structure and function, and can have a specialized role within an organism. Mechanisms...wrapped around histones changes, then gene expression can change as well. The more frequently and tightly bound...in the promoter region of tumor suppressor genes can repress their transcription and inactivate them. ...bisulfite sequencing. However none of these techniques can be used for in-situ research of dynamic modifications...

...decide to share them using Addgene’s repository, you can also share your associated protocols. When you upload.... And more plasmid-related educational resources can be found in our Plasmids 101 blog series. Want to... Addgene? Check out all the benefits and how you can get started on our deposit page. Protocols.io Website...a life science protocol repository. The software can be used with the ZappyLabs Bench Tools mobile app...to use, and add your own comments. Currently you can find protocols from NEB, various academic labs, and...allows labs to create accounts in which their members can share protcols, sign-up for shared equipment, and...software designed with scientists workflow in mind and can offer a one-stop solution to your lab's organization...

... available. Without a comprehensive resource, it can be overwhelming to choose a Cas for your experiment...planning, you need more nitty-gritty details — how can I multiplex this enzyme? What other RNA components...embedded in the References lists on every page you can find direct links to plasmids from the article if... a Cas entry (SpyCas9a) from CasPEDIA.   How can I use it? CasPEDIA operates like many search engines...For the advanced search options, this is where you can really identify the best Cas for the job! There are... are a series of drop-down menus in which you can manually enter details to refine your search (trans ...ability, gRNA design, etc.). With this feature, you can compare different Cas enzymes to make sure you are...

... kb, ample room for most experimental needs, and can transduce (infect) a wide range of cell types with... coming in around 9 kb. Unlike gamma, lentivirus can infect both dividing and non-dividing cells. Lenti...envelope or a post-transcriptional regulatory element can be used for either type of retrovirus. Adeno-associated... is a small, single-stranded DNA parvovirus. AAV can infect humans and some other mammals but is not currently...an extrachromosomal part of the host genome that can replicate and divide along with the endogenous DNA...high immune response as retroviruses. This virus can also transduce dividing and non-dividing cells, making...routinely being developed) to choose from, which can target the virus to specific cell types of interest...

... scientific and medical tools. As scientists, we can manipulate what this cargo is and where it goes, ...cells they infect. In the lab, this means viruses can be engineered to be cell type specific. Once inside...genome, however, isn’t enough to ensure the virus can replicate and spread. Instead, viruses replicate ...one they have a high transduction efficiency and can effectively infect a wide range of cell types that...difficult to transfect’, such as primary cells. They can also be directly injected into hard-to-reach organs...Lentiviruses are a genus of the retroviral family that can infect both non-dividing and dividing cells. They... this through nuclear localization signals which can traffic their viral particles inside the nucleus,...

...platforms such as Github, users can download the software for use, or they can modify and optimize the code...have been convinced that most microscopy techniques can be replicated with commercially available consumer-grade...feasibility of manufacturing, and subsequent assembly can be difficult to overcome. Universities may not have...representations of the 3D models, which in many cases can be produced quickly with the printer, act here as... since a part developed in Germany, for example, can also be printed quickly in Madagascar. Platforms ...across laboratory and disciplinary boundaries and can allow for innovative organizations and companies ...example of how sharing of data and molecular tools can lead to faster and more efficient science. Researchers...

...LentiCas9-blast virus can be used to generate Cas9-expressing cell lines, which can then be used both for...collection of tools so that researchers around the world can accelerate their work. After all, as we like to sayat...is the headliner of most CRISPR applications and can be expressed from your cell line of choice after ...nuclease, which introduces double stranded DNA breaks, can be expressed from lentiCas9-Blast. Purpose of LentiCas9...for generating your own lentivirus. Secondly, you can use a LentiCas9-Blast viral prep to create a single...University of North Carolina, Chapel Hill. These can be used to investigate the effects of activating ...Purpose of this AAV pAAV-hSyn-DIO-hM4D(Gi)-mCherry can be used to induce neuronal silencing in vitro and...

...tags can be used to facilitate the visualization of specific proteins in cells. Tagged protein can easily...specific antibody and the proper microscope setup we can observe the whole population of one particular protein...proteins instead of methionine. The labeled proteins can be detected using click-chemistry that covalently...alkyne tag. The newly synthesized and tagged proteins can then be observed with fluorescence microscopy. This...Professor Schuman’s lab showed that FUNCAT’s usefulness can be extended to determine the turnover of a specific...of interest are in close proximity to each other can the DNA strands take part in rolling circle DNA amplification...discover new restaurants, go to concerts and bake. you can contact her at [email protected]. References ...

...your experimental RNA. To solve this problem, gRNAs can be chemically modified to resist degradation while... Biotechnol; Rozners, JACS). These modifications can be done in-lab using commercially available kits ...surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency. In the same vein, ..., locked nucleic acids or bridged nucleic acids can also be introduced in the gRNA. These modifications... 20-bp targeting region of the gRNA. These gRNAs can be cleaved and rendered useless after less than a... the gRNA is released and the CRISPR-Cas9 system can initiate targeted editing events. Both technologies...There are many suitable dyes to choose from and you can opt to do-it-yourself with a kit or purchase it commercially...