We narrowed to 1,052 results for: FER

...updated by Jason Nasse in July 2023.  References and Resources References Chen X, Choo H, Huang X-P, Yang X...lead to such responses as decreased heart rate (Redfern et al., 1999) or altered neuron excitability (Zhu...to consider that the same receptor can elicit different responses depending on what cell type it is expressed....sciencemag.org/content/364/6436/eaav5282.long Redfern CH, Coward P, Degtyarev MY, Lee EK, Kwa AT, Hennighausen...

...concentrating a viral stock. Different titering methods can reveal differences in physical versus functional...will be used in a couple of days, some researchers prefer to store freshly harvested virus at 4 °C instead...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate... Yarmush. "Polybrene increases retrovirus gene transfer efficiency by enhancing receptor-independent virus...

...compare results across multiple targets because different CAPTURE cell lines will have varying levels of...lines Cell lines, primary cells Table 1: Key differences between original CAPTURE and CAPTURE 2.0 methods...that occur during development and cellular differentiation. Using a cell line model, the Xu Lab tracked...enhancer-promoter interactions during erythroid differentiation. CAPTURE 2.0 captures chromatin's dynamic role....0 re-writes the script on gene regulation. References Han, J., Zhang, Z., & Wang, K. (2018). 3C and...

...randomized with a defined set of different trinucleotides (corresponding to different amino acids) in the regions...this, they injected the spike protein from two different coronaviruses into a llama over several days and...your target protein. The Seeger lab designed 3 different mRNA sybody libraries for use in the protocol,.... Find the Seeger lab's SARS-CoV-2 sybodies! References Geertsma ER (2013) FX Cloning: A Versatile High-Throughput...

... splitting transfected cells into 3 plates at different densities for selection). A 2% agarose solution...transfection into plates containing selection agent at different densities, then dispose of the plates that do ...freeze at least 2 ampules each of 4 clones with different levels of expression of our gene of interest. ...to rapidly isolate individual cell clones with different expression characteristics from a population of...cell clones picked using the agarose method do not differ grossly from those isolated using cloning rings...

...comes to fruit flies Drosophila start life as fertilized eggs that develop into embryos and then become...about one week from the initial deposition of the fertilized egg by its mother. Inside the pupal case, the... the heat. Isolating male and female flies Differentiating male and female flies is essential for setting...thorax to look like stubble (sb). For a better reference of various fly phenotypes, check out the Classical...latest information about gene function, while others offer fly stocks and other reagents. If you’re ever confused...

...create customized overhangs to assemble up to 6 different fragments in a prescribed order. These fragments...protocol is fairly flexible and adaptable to cloning different types of DNA, such as cDNA, PCR fragments, or ...process close to 100% efficient. Gateway cloning also offers a very high chance of success (>95% for a single...deposit your finished constructs at Addgene!). References 1. Li, Chaokun, et al. "FastCloning: a highly...Find vector backbones Read our molecular biology reference ...

...these vectors is controlled by a different promoter resulting in different levels of expression from each...functional, stable proteins. Protein expression refers to the second step of this process: the synthesis...well-defined media and can be easily adapted to fermentation allowing for large-scale, stable production ...labelled or modified amino acids that are useful in different downstream applications. Cell-free expression ...

...library to identify RBD mutations which do not interfere with ACE2 binding, but impact recognition by specific...immunoprecipitate the entire complex in three different cell lines. The Gräslund lab plans to continue...10.1101/2021.01.30.428974   The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR...of indels to measure time. These plasmids with different promoters driving expression of Cas9 were used...Janelia Farm GENIE project have arrived! We now offer several versions of GCaMP8 vectors in serotypes ...

... Let it incubate Check the bacterial strain Differences between E. coli host strains can impact plasmid...might want to include an extra wash step or use a different strain. For most applications, Addgene uses and...inadequate plasmid DNA yields. Why would volume make a difference? In the simplest of terms, more volume = more...conditions play a critical role in plasmid DNA yield. Different plasmids and strains will vary in their optimal...media with supplements such as magnesium salts, buffering agents, and/or supplying additional carbon sources...

... alter the ability of one FP to nonradiatively transfer energy to the second FP, often decreasing the ...most cases, by using monomeric FPs or FPs from different families, which cannot form heterodimers (i.e....University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes...avid long distance runner, gardner, and cook.   References 1. Snapp, E.L., et al., Formation of stacked ...Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...

... ever found yourself bamboozled by all of the different kinds of information on our plasmid pages? Well...depositing lab, although you may have an entirely different use in mind! This section of our plasmid pages... you to use in your Materials and Methods and References sections when citing the use of your colleague...backbones.  The backbone modifications listed can refer to changes made to the original backbone by the ...e.g. if it was derived from a plasmid made in a different lab - or one obtained from Addgene!).  It also...

...600 bp) and may take up precious space in viral transfer vectors with limited packaging capacity. Additionally...placement of the reporter gene). Addgene's collection offers a variety of plasmids to express two or more genes...Retroviral pEF1a-IRES-Neo IRES Mammalian MSCV-IRES-Luciferase IRES Retroviral pWPI IRES Lentiviral AmCyan-P2A-mCherry... new eBook! Our goal was to create a one-stop reference guide for plasmids. We've combined our Plasmids...solutions necessary for advancing the field. References: 1. Internal initiation of translation of eukaryotic...

...CRISPR gene activation system, but settled on a different optimal fusion protein combination. The optimized...AcrIIA4, this unfolding conformational shift can interfere with activity. The resulting system was coined...Alexander Deiters’ lab, on the other hand, took a different approach. These researchers used a genetically...expression - demonstrating light-induced silencing of transferrin receptor CD71 in HeLa cells.   Comparison of...applications you use these for in the future! References Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner...photoactivatable transcription systems to induce neuronal differentiation. Nat Methods 14:963–966 . https://doi.org/10.1038...

...twitter @Vopinator.     References: Halpin-Healy TS, Klompe SE, Sternberg SH, Fernández IS (2020) Structural...Insertion of the mini-transposon can occur in two different orientations, but is strongly biased towards T-RL...combined in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and the...integration efficiencies achieved with VchINT, at two different incubation temperatures. Data adapted from Vo ...

...the same figure by figure organization with a different person responsible for each figure. Others still...questions they have.  These variations are suited for different applications and should be carefully chosen for... and audio recordings of the meeting using teleconferencing software. Any file sharing service should ...or instant messaging but we as scientists often prefer data. At Addgene we rely on polls and surveys. ...acquisition of the essential data and allow everyone to offer thoughts you may not have even considered. These...

...written by Addgenie Brian O'Neill.  References and resources References Bauzon, M., Castro, D., Karr, M.,...vectorization strategies include creating an AdV transfer plasmid with the E1 and E3 (Early 1 and Early ...Although the late genes are usually present in the transfer plasmid for the majority of adenoviral vector ...B. (2014). Adenoviruses: General Features. In Reference Module in Biomedical Sciences. Elsevier. https...

...This intrinsic bacterial recombination pathway, referred to as recA-independent recombination, joins together...single PCR, for example for subcloning genes from different templates. All PCRs are run as an 18 cycle, 25...pre-made aliquots of PCR master-mix, containing buffer, polymerase and dNTPs, can be stored frozen: just... on Twitter @jakefwatson and @JGarciaNafria.  References 1. Jones, Douglas H., and Bruce H. Howard. "A...for plasmid cloning Find our molecular biology reference here Find molecular biology protocols here   ...

... in and out of the classroom.   References and Resources References Bangera, G., Brownell, S. 2014. Course-based...the University of San Diego), decided to take a different approach. His own most impactful learning experiences...all shaped his teaching style, with the former offering excellent training in improvisation and the latter... available technical and pedagogical resources, offers mentoring for instructors, and provides opportunities...

...denotes intersection and the – operator denotes set difference. That’s the intuition for how genome math helps...enhancers from step 2. See which ESCs die or differentiate Step 3 reveals which NANOG-related genes are...distinct enhancer disrupted. As our ESCs die and differentiate over time, we periodically use next-generation...Therefore if a gRNA drops or disappears over time, we infer that the enhancer it targets is a “pillar of function...by starting with all of the DHS sites from 125 different human cell types from the ENCODE Project. DHS ...

...viral infection), producing firefly luciferase. Firefly luciferase can readily be measured to quantify...single (non-viral) reporter gene (e.g. firefly luciferase or eGFP) flanked by the EBOV-specific gene start...to transcribe the reporter gene (e.g. firefly luciferase). The polymerase recognizes the gene start signal...transcription of negative strand RNA viruses.   References 1. Buchholz, Ursula J., Stefan Finke, and Karl-Klaus...

...human pluripotent stem cells By Jennifer Tsang CRISPR gene interference (CRISPRi) and gene activation (...For labeling the plasma membrane, they tested different target sequences for improved selectivity and ...into the supernatant. Four plasmids containing different nanobodies (NB) were generated in pMAK:  pMAK461....1038/s41598-020-57500-1 The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR...

...limitations associated with SpCas9. Cas13a (previously referred to as C2c2), has several unique properties that...for target recognition and cleavage). C2c2 (now referred to as Cas13a) is unique in terms of structure ...remainder of this blog post. Perhaps the biggest difference between Cas13a and Cas9 is that Cas13a binds ...SHERLOCK (depicted in figure 1) has been used to differentiate strains of Zika virus, genotype human DNA, identify...Gearing contributed to the update of this post. References 1. Abudayyeh, Omar O., et al. "C2c2 is a single-component...

...moving around to explore the environment has a different motivation (and brain areas associated) compared...addiction. Anxiety tests Rodents have a natural preference to stay in dark areas (protected) as opposed ...to brightly illuminated ones (risky). They also prefer to explore novel environments by initially moving...anxiety. On the other hand, the control group preferred staying in the dark compartment (Howerton at al...these in a  future blog post, so stay tuned!  References Alpár, Alán, et al. "Hypothalamic CNTF volume...

... to determine the pros and cons of using many different types of gRNA design software. When you are happy...inserted or deleted. Each lab will use a slightly different method to genotype their pups. Taking a toe, a...weeks old and use the ear tag method (creating different patterns of ear holes and using that material ...latest and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target...plasmid into zygotes." Development, growth & differentiation 56.1 (2014): 122-129. PubMed PMID: 24372541...

...labs. While many labs approach the situation differently, some adopting simple note-taking tools, others...functionalities with experiment records. Many different types of ELNs are available on the market today... comparison to paper-based record keeping, ELNs offer functionalities that are hard to imagine on paper...desired functionalities i.e. main priorities of different ELN features, ranging from whether respondents... allows you to assign specific permissions to different lab members to avoid any issues later on. Do you...

...delivering scFvs that inhibit HIV.   References and Resources References: Ahmad ZA, Yeap SK, Ali AM, Ho WY...for high affinity scFvs. Bound phage are then proliferated by infecting bacteria for additional rounds ... high affinity binding. Bound phage are then proliferated by infecting bacteria for additional rounds ...science.3140379  Bradbury ARM, Sidhu S, Dübel S, McCafferty J (2011) Beyond natural antibodies: the power...

...C-terminus of the fluorescent protein. I prefer to describe the different parts of the fusion from N- to C-terminus...tagged protein because of its size. This type of interference is known as steric hindrance. For instance, ...advisable to generate multiple constructs with different insertions, since it is difficult to predict which...can follow him on twitter: @joachimgoedhart.   References Adjobo-Hermans, Merel JW, et al. "Real-time visualization...

...interviews, or get several interviews but no job offers, is a failure to clearly articulate one's talents...to convince me that their search committee is different and needs to be told all of the nuances of the...technologies, i.e. video interviews. For point of reference, I submitted my applications all by snail mail...hoppy pint of IPA or a few drams of single malt (preferably from Islay or Skye). Good luck with your search...University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes...

...of S. pyogenes Cas9 or Cas9 variants (hereafter referred to as Cas9) and a single stranded guide RNA (gRNA...agrobacterium-mediated delivery. Summary While different in many of the specifics - promoters used, precise...mediated genome engineering in plants isn’t all that different from how it’s used in other systems. Luckily, ... don’t be afraid to try your hand at CRISPR. References Belhaj K, Chaparro-Garcia A, Kamoun S, Nekrasov...of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew. Nat Biotechnol...

... Even low-frequency editing could make a huge difference in DMD, and it’s estimated that exon skipping...respectively. Although each study used slightly different methods, they each observed the same positive ...disease type C. Although the three DMD studies referenced here represent a great step forward for CRISPR...many distinct CRISPR therapies to match these different mutations. For each therapy, effectiveness and...treatments may also be closer than we might expect.   References 1. Kaiser J. CRISPR helps heal mice with muscular...

...bottom of this post for a basic synopsis of the differences amongst the Brainbow systems. Pre-Brainbow techniques...to the promoter will be expressed. This system differs from Brainbow-1.0 in that transient Cre recombination...needed to mark individual neurons, the system also suffered from a number of limitations. First, the imaging...controlled by FLP/FRT recombinase. When placed under different promoters, Brainbow and Flpbow can be used to ...2.1 plasmids Brainbow 3.0, 3.1, 3.2 plasmids References: Transgenic strategies for combinatorial expression...

...PiggyBac transposons, the Woltjen Lab compared different variants of the polycistronic cassettes and discovered...a donor in FRET (fluorescence resonance energy transfer) and FLIM (Fluorescence Lifetime Imaging Microscopy...is a new member of this class that works via a different mechanism - the PhoCl protein is irreversibly ... are other RFP variants available that may be preferred for their intended uses, they provide a well-defined...of essential genes in vivo. Utilizing CRISPR interference (CRISPRi) technology, the arrayed knockdown ...defined domains can be swapped onto other proteins to confer functionality. In Narayan et al., they used this...

...Szymanski in January 2016 in Viruses (5). Please refer to the paper for more details on the assay, including...however, a physical fragmentation strategy is preferable. Nebulization involves forcing DNA through a ...on the membranes will then be exposed to a lysis buffer to release recombinant proteins directly onto the...then suspend a freshly harvested dose of cells (preferably in log phase) in blocking solution (around 108...thinking. Find her on Twitter @JessicaSacher.   References 1. Nobrega, Franklin L., et al. "Revisiting phage...

...Learn about different molecular cloning techniques, focusing on seven common cloning methods....Educational Resources Molecular Biology Reference Molecular Cloning Techniques Molecular Cloning Techniques...availability of starting materials, or just personal preference. Check out our blog on choosing the right cloning... entry , and Gateway destination vectors with different promoters, tags, selection markers, and fluorescent...

...“NCBI Site Preferences” box. Once on the Preferences page, navigate to “PubMed Preferences” and click ... that you know how to navigate to the PubMed Preferences and Filters pages, be sure to check out the other...

...Request service, which offers over 3,000 eligible transfer plasmids and five different serotypes. Not sure... pAAV_hSyn-PdCO-mScarlet-WPRE AAV1 Optogenetics Ofer Yizhar New viral tool pAAV-Ef1a-fDIO hChR2(H134R...

...modified allele is selectable, it can be transferred to a different genetic background via P1 transduction...Figure 2). In the following sections, key steps that differ from traditional restriction enzyme cloning will...could be used to move the modification into a different background, if needed. This approach is best suited...science communication and in vitro diagnostics.     References 1. Costantino N, Court DL. Enhanced levels of...

...AAV constructs CRISPR interference in Candida albicans A new CRISPR interference (CRIPSRi) platform for... to Cas9 SECURE base editors Nme2Cas9 CRISPR interference in Candida albicans E. coli genome-wide inhibition... Jennifer Tsang...

...animals’ skeletal muscles led to another clue: differences in the kynurenine pathway of tryptophan degradation... skeletal muscle expression of kynurenine aminotransferases, to enhance the conversion of kynurenine into...Get started with PGC-1 plasmids from Addgene. References: Skeletal muscle PGC-1α1 modulates kynurenine...M, Bhat M, Schuppe-Koistinen I, Pettersson AT, Ferreira DM, Krook A, Barres R, Zierath JR, Erhardt S, ...

...available at Addgene CRISPR-Cas9 References, Protocols, and more References: Generation of Gene-Modified ...the monkey story hit the presses, another very different bit of CRISPR news was circulating. Scientists... work. Last month, the Lawrence Berkeley Lab's Jennifer Doudna and Eva Nogales presented the first detailed...

... online. A few examples are provided here for reference: Untergasser Lab Preparation of chemically competent...use the Mix & Go! E. coli Transformation Kit and Buffer Set  from Zymo Research to make competent cells...then wash and resuspend the cells in the provided buffers. The competent cells are then aliquoted and stored...protocol)  Cells prepared using the Z-competent TM buffer can be easily transformed without a heat-shock ...

...finding the best way to organize your time. At a conference, you could be honing your science communications...that lab and interacting with the lab members.”- Jennifer Tsang, Science Communications and Marketing Coordinator...Addgene blog Read all careers blog posts Read our transferable skills guide Resources on Addgene.org Find... Jennifer Tsang...

...shipped over 350,000 individual plasmids to 5,000 different research institutions. This has given us a unique...by the influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids...HA-Ubiquitin-WT and HA-Ubiquitin, and we now have 19 different ubiquitin plasmids in the repository. 100,000 ... and modified by so many labs and for so many different purposes that we had to create a CRISPR page to...

...the researchers applied the LITE system to 28 different genes to establish it as “a novel mode of optogenetic...activators are easy to expand or combine with different effectors. I’d encourage people to get creative...apply LITEs for the transformation of neurons, she offers a tip: save yourself some time by making an AAV1...readers, how will you get creative with LITE? Reference: Silvana Konermann et al. "Optical control of ...

...inflammatory cytokines - interleukin (IL)-12 and interferon gamma (IFNγ) - and eliminate cancer cells. 2....physiological and morphological changes. Tumors differ in their genotypic and phenotypic identities based...by cancer. As mentioned in this article, cancers differ in practically every facet starting from manifestation...manifestation to their specific interactions with different cell types. A thorough understanding of the genotypic...interested in exploring new avenues in healthcare.   References 1. Vinay, Dass S., et al. "Immune evasion in ...

...York City, NY, USA 40.7128°N 74.0059°W – Deanna McLafferty I’m a New Yorker, so it’s not easy to convince...myths that abortions cause breast cancer and infertility or to argue against unnecessary clinic regulations...and fiction. Our health depends on it. Deanna McLafferty is an editorial assistant and activist. Washington...enough.  Nashville, TN, USA 36.1627°N 86.7816°W- Jennifer Stancill I am a research biologist by training...supporters is much larger and stronger that I expected. Jennifer Stancill, Ph.D., is currently a post-doctoral ...

... and Negative Selection for Plasmid Cloning by Jennifer Tsang There are many strategies for finding your...Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi by Jennifer Tsang Learn about mNeonGreen, mTFP1, and mWasabi...Precision and Flexibility to CRISPR Editing by Jennifer Tsang Prime editing is a new CRISPR technique... Jennifer Tsang...

...simplifies your viral vector options. Almost any AAV transfer plasmid in our repository can now be packaged ...packaged on demand in five distinct serotypes, offering you unparalleled flexibility. But this is just the beginning...developments? Subscribe to the Addgene newsletter. References Ayoubi, R., Ryan, J., Gonzalez Bolivar, S., Alende...A., Trimmer, J. S., Rego, M., Freedman, L. P., Ferrara, F., Bradbury, A. R. M., Cable, H., & Longworth...

... might not fold well, you should screen a few different vectors to see which gives you the most functional...two sets, each of which includes plasmids with different varieties of tags, both N- and C-terminal. Each...improve folding and disulfide bond formation. If one prefers expression in E. coli, the use of a thioredoxin...individually or as a complete kit from Addgene. References 1. Goulas, Theodoros, et al. "The pCri system...