We narrowed to 321 results for: POR C

...our guest, Darrell Kotton. Dr. Kotton is the David C. Seldin Professor in the Department of Medicine and...iPS cells derived from patients. Chapter 8: The importance of a mix of research approaches. Why researchers...

...site). Finally, in addition to the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X FLAG tag at...are some additional features. The first and most imporant is that, upstream of the Cbh promoter, this plasmid...

...reaction takes an hour or less to complete at 50 °C. After an hour or so, the sample is immediately ready...PMID: 25861928. 3. Jiang W, Zhao X, Gabrieli T, Lou C, Ebenstein Y, Zhu TF. Cas9-Assisted Targeting of CHromosome...primers for PCR amplification of fragments to incorporate such regions of homology. This video gives a ...

...ask at least one question to clarify the situation; C = confirm that you have understood the speaker accurately...focus on an aspect of communication especially important for a manager – getting feedback from the team... This can take practice and training. This is important because it creates a culture of engagement in ...  Seeking feedback frequently One of the most important paths of information flow is the one that brings...

...fluorophores with nonoverlapping emission spectra (N- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus, ... of transcription factors OCT3/4, SOX2, KLF4, and c-MYC is the established model for reprogramming somatic...coselection, analogous to methods previously used in C. elegans, targets the sodium/potassium pump ATP1A1...phosphatase-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked mCherry...are available at Addgene. Kim, et al. Stem Cell Reports. 2015. PubMed PMID: 25772473 Targeting and manipulating...Genetically modified organisms like transgenic mice are important tools for understanding the function of cell types...enhancer elements and combined it with a variety of reporters (i.e. eGFP) and effectors (i.e. DREADDs). The ...

..., G. S. Parker, S. J. Qualls, B. M. Lewis, E. Ke, C. T. Dargitz, R. Wright, A. Khanna, F. H. Gage, and... Mikkers, J. Kool, L. van der Weyden, A. H. Lund, C. H. Wilson, R. Rance, J. Jonkers, M. van Lohuizen,...useful for genetic engineering approaches. Most importantly for this technology, however, is that the transposase...should be tested in the cell line of interest. Importantly, by varying these ratios, you can either maximize...obstacle, a positive selection cassette can be incorporated, and used during the differentiation process...homologous recombination, which results in the incorporation of your specific edit into the host genome. ...transposon sequences.  The use of PBx is particularly important to avoid any possible random re-integration from...

...little to no signal in the liver sample, which you do (C), so your antibody is validated! Genetic Strategies...10.1080/19420862.2015.1100787  Taussig MJ, Fonseca C, Trimmer JS (2018) Antibody validation: A view from...Reproducibility First, let’s review why validation is so important. Antibodies are a critical component of the researcher...or plan your own validation experiments, it’s important to know what you are aiming to achieve. In general...as whole cell lysate or tissue sections. It is important to confirm that your antibody is not binding to...proteins bound to the target protein, so it’s important to understand if protein:protein interactions ...

...Plans Aren't Just More Paperwork. Vincent BJ, Scholes C, Staller MV, Wunderlich Z, Estrada J, Park J, Bragdon...and PI, it’s easy to miss out on beneficial opportunities or to find oneself unsure of the next steps....planning meetings with each trainee to create a supportive, growth-oriented lab culture. The aim of these...the overall tone of the meeting is almost more important. As DePace says, science is full of “terrible ...postdoc training. On the other side, the PI has an opportunity to check in with lab members, determine how to...

...PubMed Central PMCID: PMC1559948. Brusca, Richard C., and Matthew R. Gilligan. "Tongue replacement in ... please! At Addgene, we love finding ways to incorporate science themes into our fun activities and Halloween...body. The fruiting bodies then rupture and release spores onto the forest below for the next unsuspecting...

...Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y, Lin C, et al. An erythroid enhancer of BCL11A subject to...work was with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began...correlative biochemical characteristics and ectopic reporter assays. We have been interested in exploring the...transfect CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via FACS, primer...

...Addgene.   References Ke, Q., Ahn, Y., & Sugimoto, C. R. (2017). A systematic identification and analysis...physical scientists are underrepresented. It’s important to note that these results could be biased for...domains more often than other scientists   Figure 5: Portion of Tweets with Scientific Domains, Categorized ...

...attach the gene of interest to either the N-or the C-terminus. Even if a researcher’s gene of interest ...reagents. "We are proud to be able to bring such an important collection to the scientific community," says ...scientists, and the FSU team who have made this large, important plasmid deposit possible.   ...

...annurev-pharmtox-010814-124803 Vlasov, K., Van Dort, C. J., & Solt, K. (2018). Optogenetics and Chemogenetics...   Timing Optogenetics offers a high degree of temporal resolution as receptors can be turned on and off... meet these requirements. TL;DR If precise spatiotemporal control of a small subset of neurons is necessary...

...proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins. While the unique...capsid, “pumps” the genome into the capsid through a pore in the capsid. Rep proteins are also involved in...2020) Adeno-Associated Virus Genome Interactions Important for Vector Production and Transduction. Human ...

...transfection. This supernatant can be stored at 4 °C or can be directly added to your target cells. However...and plasmids.  References 1. Miller, A. DUSTY, and C. A. R. O. L. Buttimore. "Redesign of retrovirus packaging...or electrical (electric current forming membrane pores) transfection. These techniques are still used and...has been removed, Rev (which facilitates nuclear export) is expressed from a separate plasmid, and the ...you clone your gene of interest into and is an important feature for your experiment; it must be chosen...using them.   Regarding biosafety, it is also important to be aware that lentiviral particles are a powerful...

...PubMed Central PMCID: PMC4349094. 3. Bell, Charles C., et al. "A high-throughput screening strategy for... 22963480. PubMed Central. 5. Tsiatis, Athanasios C., et al. "Comparison of Sanger sequencing, pyrosequencing... the University of Cambridge. One of the most important steps in the CRISPR experimental process is validating...silico prediction. These techniques differ in important ways, but can also complement one another by providing... unbiased methods, high background BLESS Not reported Cell-based (in vitro) Can be used on tissue from...senstive to time since cell fixing HTGTS Not reported Cell-based Identifies translocations Limited by...

...neuronal phenotype in a mixed population (Burger, C. 2004, Aschauer, DF. 2013, O’Carroll, SJ. 2021). This...https://doi.org/10.1371/journal.pone.0076310 Burger C, Gorbatyuk OS, Velardo MJ, Peden CS, Williams P, Zolotukhin...fibrillary acidic protein (GFAP) for astrocytes it is important to test it in the brain region of interest, especially...of Transduction Efficiency, Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse... the Addgene blog Plasmids 101: FLEx Vectors Important Considerations When Using AAVs Three Key Considerations...

.... DOI: https://doi.org/10.1080/rsob.180241 Vanni, C., et al., Unifying the known and unknown microbial...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as... screens to introduce reporters throughout an organism’s genome. These reporters have historically been...stimulate the expression of the reporter. The system is made up of a reporter gene flanked by a minimal promoter...promoter – all rely on the random integration of a reporter construct in close proximity to an endogenous ...example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed in the kidneys... for with an uncharacterized gene. The actual reporter construct can be something phenotypic (such as...

...of our most popular mammalian expression vectors, c-Flag pcDNA3, comes from the pcDNA series. It has the...consider as you make your choice. The first and most important thing you need to know is your expression system...have to make sure that your plasmid has been incorporated into the host organism, usually achieved with...basal expression level, induce expression at an opportune time or go all out with constitutive high level...Some of the more popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. ...animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g...

... Deverman BE, Matho KS, Cetin A, Woodard K, Cepko C, Guerin KI, Rego MA, Ersing I, Bachle SM, Kamens J...’s ~4.4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such...the Cre recombinase, are used to spatially and temporally control transgene expression. Lox sites are directional...sites lets researchers control the spatial and temporal activation of a transgene. Figure 2: FLEX...structure when transcribed which aids in nuclear export of mRNA and helps increase the expression of your...polyadenylation or polyA signal aids in the nuclear export of RNA and RNA translation, and promotes RNA transcript...

...Boeckman JX, Manning CF, Nguyen JT, Bechtold H, Dumitras C, Gong B, Nguyen K, van der List D, Murray KD, Engebrecht...and have always been, a community resource that supports both toolmakers and tool users. We encourage you...Educational Resources As part of our commitment to supporting your research, Addgene is creating an array of...Through our incredible community, including your support, we have enabled scientists to easily share plasmids...Studies Hybridoma Bank, a nonprofit that has been supporting hybridoma sharing within the community for decades...Immunofluorescence Addgene’s Antibody Guide The project is supported by a NIH BRAIN Initiative grant (U24NS119916 )...

...∆G, however, can not be conjugated to PE due to a C-terminal glycine deletion, so it remains in the cytosol... development of several fluorescent autophagy reporters now allows for the measurement of autophagic flux...makes Keima useful for monitoring mitophagy. It’s important to remember that because Keima’s fluorescent relies... al. "Rosella: a fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in...using a single fluorescent biosensor." Scientific reports 5 (2015): 12397. PubMed PMID: 26215030. PubMed ...

... antibodies aren’t free to bind endogenous IgGs. (C) A third solution is to use Fab fragments against ...primary antibody that is directly conjugated to your reporter circumvents the issue. However, directly conjugated...which uses just a primary antibody that has your reporter (in this case a fluorophore) directly attached... as effective for other techniques, but it is important to be aware that the issue is not unique to IHC...

...under the United States Internal Revenue Code 501(c)(3). That means we were formed to benefit the public...do. From all of us to all of you. Thanks for supporting nonprofit, open science and Addgene in our mission...Collaboration with Fellow Nonprofit Seeding Labs Addgene Supports Rare Disease Research Listen to Our Interview ...

...released from nuclei by a short incubation at 37 °C. These short DNA fragments can then be purified for...MNase to antibody bound proteins (Figure 1). Importantly, since Protein A has weaker affinity for mouse... rather than to rely on nuclei extraction. They report that this change offers a significant improvement...

...) antibody-dependent cellular cytotoxicity (ADCC) c) antibody-dependent cellular phagocytosis (ADCP). ...starting a cascade of events that results in the pores being opened in the pathogen’s cell membrane (Dunkelberger... when holes are punched in their membranes, the pore'd pathogens quickly lyse and die.  Antibody dependent...

.... Scientific Reports, 10(1), 21842. https://doi.org/10.1038/s41598-020-78827-9  Breen, C. J., Raverdeau...Muthusamy, A. K., Liang, Y., Brown, T. A., Lemon, W. C., Patel, R., Lu, R., Macklin, J. J., Keller, P. J....protein function and cellular components: It's important to assess how dye-conjugation affects protein ... interpretation of experimental results. It's important to consider how dyes can influence receptor conformation...efficacy of these ligands can be affected by the incorporation of fluorophores, as seen in studies involving...wavelengths for green, red, and far-red dyes is important for intracellular fluorescence live imaging ​​(...from cells under certain conditions, which is an important consideration for assay design (Breen et al., ...

...T-regs) into their microenvironment by producing C-C motif chemokine 22 (CCL22) and TGFβ. T-regs are suppressive...still many important questions in this exciting field of research. One of the most important: How will ...commute to and invade multiple organs, and most importantly escape the immune mechanisms of the host. The...against cancer. Sipuleucel-T by the Dendreon Corporation is currently the only FDA-approved DC therapy...therapies themselves can be made more effective. Some important factors researchers are beginning to focus on ... of cell types. This cell heterogeneity is an important element of successful tumor progression. A tumor...), and ability to metastasize. There have been reports of clinical trials going amiss and patient deaths...

..., larval zebrafish react by performing a distinct C-bend to escape possible danger (see figure). Larvae...: 19805086. PubMed Central PMCID: PMC2764931. Portugues, Ruben, et al. "Optogenetics in a transparent ...

...allow the digestion to take place for an hour at 37°C. This should be enough time to completely digest the...overhangs, insert and vector purification, and, most importantly, excessive time requirements. FastCloning is ...

... Kunjapur, Aditya M., Philipp Pfingstag, and Neil C. Thompson. "Gene synthesis allows biologists to source...is evolving so fast that it is a really nice opportunity to study it and see what happens when technological...

... new scientific observations. References 1. Rago C, Vogelstein B, Bunz F. Genetic knockouts and knockins...Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE. Nat ...of the gene with multiple gRNAs, but the most important is that you could avoid potential off target artifacts...you could run another gel that highlights the important results (for example, if you needed to make a ... Despite the shifting landscape of research opportunities, it’s an exciting time to be in the lab. Based...

...protein of interest with a fluorescent reporter on the N- or C-terminus of the protein. One downside of...doi.org/10.1002/cpim.40 Picot J, Guerin CL, Le Van Kim C, Boulanger CM (2012) Flow cytometry: retrospective...a pressurized buffer called sheath fluid and transported through tubes or capillaries to a laser. As the...

...biochemistry. There, I had exchanged contact with Amy C., Addgene’s intern coordinator shortly after hearing...capstone project report and presentation, during the final week at Addgene. This report shared my work ...Coming from an environment with limited opportunities and support, puts into perspective just how much one...hearing her talk about Addgene’s internship opportunity taking place next summer.  I applied to Addgene ...to differentiate which clone had successfully incorporated Syntaxin3 into its backbone. Once we had gotten...

...light. Previous iterations of FLiPs only used N- and C-terminal dronpa fusions. With these various designs...back in with the Addgene blog for future award opportunities. Xin Zhou: Fluorescent Light-inducible Proteins...particularly notes their ability to be precisely spatio-temporally controlled as a key advantage over other inducible...Oligosaccharides (HMOs). HMOs are thought to play an important role in nuturing the development of beneficial...consisting of sfGFP fused to a His-tagged ABC transporter known to bind to the HMO of interest. Fatima ...

...Antonini, M.-J., Rosenfeld, D., Park, J., Garwood, I. C., Yan, W., Niroui, F., Fink, Y., Chandrakasan, A.,...included in them for downstream control – this is important later! Next, additional components are added -...times more flexible than the brain fibers and importantly distortion of the fibers didn’t affect their ...temperature proximal to the implanted fibers. As important as its capabilities are the practical considerations...

... I as this enzyme recognizes the DNA sequence 5´-(C/T)CCTT-3' and digests double stranded DNA at this ...salt causes arcing during electroporation which would cause the electroporation to fail). When incubating...limit (lower transformation efficiencies have been reported with longer incubation); however, you may need...

...Gelsinger, D. R., Vo, P. L. H., Klompe, S. E., Ronda, C., Wang, H. H., & Sternberg, S. H. (2024). Bacterial...bacterial genome engineering tools, with early reports of 40%–60% efficient integration (Klompe et al....streamlined, one-size-fits-most approach. evoCAST is an important step toward that kind of gene treatment. The researchers...

...PubMed Central PMCID: PMC6070312. 2. Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent synthetic...encodes it. That DNA sequence then needs to be incorporated into a host cell (through a process called transformation...FD-CF reactions affordable to many schools. Most importantly, experiments for FD-CF reactions can be fun and...

...Lindenbaum P, Martin-Fontaine M, Broucque F, Robin C, Hebben M, Merten O-W, Blouin V, François A, Redon...sequencing and analysis approach can be easily incorporated into standard QC regimens and will provide scientists...

... system is capable of robust expression of high G+C content genes, as validated through monitoring the...plasmids use “Cas9-mediated adenosine transient reporter for editing enrichment”, or XMAS-TREE, to identify...

...on designing and using RNA scaffolds: Delebecque, C. J. et al. Designing and using RNA scaffolds to assemble... source library of characterized aptamers and supporting publication: http://www.freebase.com/base/aptamer...

...arrays (e.g. for spacers A, B, and C, you could obtain arrays A, B, C, AB, AC, BC, and ABC). Nielsen Lab...expressed, the gRNAs cannot be released, adding temporal and/or spatial control to the system. pSQT1313...plasmids, which contain various promoters and reporters, and subsequently inserted into a Cas9-containing...-in-one CRISPR/Cas9 vector system.” Scientific Reports 4 (2014): 5400. PubMed PMID: 24954249. PubMed Central... Find plasmids from this paper at Addgene. 10. Port, Fillip, et al. “Optimized CRISPR/Cas tools for ...

...to produce and distribute virus. Addgene is a 501(c)3 nonprofit and our plasmid service was started based...milestone. Viral service supporters We would like to thank the companies who supported this project with funding...and be ready to provide extensive logistics support for export and shipping on dry ice. The Business and...seed money was used to build a completely self-supporting service so that Addgene is not dependent on grants... in accelerating the Viral Service. With their support we are able to provide faster access to a larger...number of viral preps. Visit our Sponsors and Supporters Page Thank you to all of our generous, sharing...Addgene repository. Without these scientists and the support of their technology transfer offices, Addgene would...

...this may be due to different lipid compositions of C. elegans membranes, which might beneficially influence...ApoB-NanoLuc reporter by inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter, the...Cell, 2019. PubMed PMID: 30639098. LipoGlo: A reporter for measuring ApoB bound lipoproteins Article ...cells. These changes were larger than previously reported for other organisms, and the Gottschalk group ...

... Cas9 for easy genome engineering in species like C. elegans and Drosophila. Light inducible promoters... phage promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, ...preventing repressor cI synthesis and allowing reporter gene transcription to take place. Addgene also...promoter is right for me? Here’s a list of some important variables to consider when choosing an inducible...’re expressing a gene that may be toxic, it’s important that your inducible promoter not be too leaky....

...Boeckman JX, Manning CF, Nguyen JT, Bechtold H, Dumitras C, Gong B, Nguyen K, van der List D, Murray KD, Engebrecht...about the process that was used to create this important collection.  Creating the Plasmids To begin constructing...antibody (R-mAb) expression plasmids, allowing for important reagents that could have been lost to continue...sequences available from each plasmid webpage. Importantly, the methods developed by the Trimmer Lab for...why choosing the right secondary is of primary importance. PLoS One. 2012;7(6):e38313. doi: 10.1371/journal.pone...

...depositing can be grown using standard conditions (37 °C, LB, antibiotics, etc.) and stored as glycerol stocks...drop-down menu to indicate whether the tag is on the N or C terminus and whether it was present in the backbone...be entirely verified by sequencing. Upload any support files that will make it easier for scientists to...

...    References and Resources References Drummond, C., & Fischhoff, B. (2017). Individuals with greater...how it works. I used to stay up past my bedtime, poring over my grandparents’ medical textbooks by torchlight...to all the optional sexual health sessions and reported the intel back to my shyer classmates. For my ...the community During my PhD, I’ve also found opportunities for more hands-on science communication. Between...

... tracrRNA and Cas9, bind to these transcripts and C) cleave them leaving mature crRNAs bound to the Cas9... combined with the tracrRNA forms the scaffold portion of a gRNA and the “spacer region” forms the target...