We narrowed to 814 results for: Ran

...false positive colonies. Transformation - the linear fragments are transformed into E. coli and plasmid...assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation. However, while...using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As this approach requires no special... encode homology arms to allow immediate co-transformation with linearized vector backbone. DpnI digestion...failed so far. What is most important is the transformation efficiency of the strain: when asking the cell...high fidelity DNA polymerase This will minimize random errors during PCR: we recommend Phusion or Q5. ...investigating the molecular biology of synaptic transmission and plasticity.     Javier García-Nafría is ...

...dCas9 has focused on transcriptional activation using a fusion to a transcriptional activator such as VP64...repression of transcription through binding a promoter region to inhibit association of transcriptional activators...locus of interest, the promoter region of the transcription factor IRF-1. Immunoprecipitation, using an ...2018). Furthermore, the Fujii Lab generated a transgenic mouse line and retroviral expression system for...Tapscott Lab identified regulators of DUX4, a transcriptional regulator that when inappropriately expressed...PMCID: PMC6001023. Fujita, Toshitsugu, et al. "Transgenic mouse lines expressing the 3x FLAG‐dC as9 protein...CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes." Cell 154.2 (2013): 442-451. ...

... inducible translocation trap (ITT) system to analyze signal-induced nuclear translocation of proteins...functions such as epigenetic regulation and transcription by using locus-specific chromatin immunoprecipitation...biochemical analysis of genome functions such as transcription and epigenetic regulation. To elucidate molecular...Addgene realizes the One-Stop shopping for a wide range of plasmids. The merit of depositing plasmids to...considering the peer-review system of journals and grant applications, for example. However, at the same ...living for Japanese, which is not trivial, and the grant situation is currently much better than that in ...tough competition for academic positions and poor grant situation. So, young researchers need to publish...

...injections can be found here [5, 6]. Opsin transgenic mice Transgenic mouse lines have also been engineered...specific brain region can result in a particular neurotransmitter release or post-synaptic current. Fortunately...available, instructions for obtaining a material transfer agreement, and list of vendors (such as the vector...at a similar spectrum as HR and also results in membrane hyperpolarization but unlike HR, this is achieved...[10]. Unfortunately, opsins are not actively transported and must passively diffuse so, depending on the...axonal tracing with fluorescent proteins in nontransgenic and cre driver mice. Current protocols in neuroscience...et al. A toolbox of Cre-dependent optogenetic transgenic mice for light-induced activation and silencing...

... after each transformation and/or use different E. coli strains with higher transformation efficiency....vector with two other independently regulated transcription units. Created with BioRender.com.   Planning...MoClo kit through their online catalog pages. We guarantee this will save you a lot of headaches down the...robot, you can set it up to automate the reagent transfer and the restriction–ligation reactions in a 96...colonies) after each consecutive assembly and transformation round, due to the decrease in ligation efficiency...’re cloning in high-throughput, we recommend transforming into chemically competent cells in a 96-well...have what look like positive colonies on your transformation plates. However, even though the MoClo system...

...protease allows a transcription factor to enter the nucleus and modulate transcription of the mCitrine ...HeLa cells were transfected with plasmids encoding FPs tagged with the plasma membrane targetin sequence...non-specific labeling. For labeling the plasma membrane, they tested different target sequences for improved...expression can vary significantly depending on transgene design and delivery. To overcome these variations...into genomic DNA with the help of the piggyBa transposase. Image from Hazelbaker et al., 2020. The lab...activating expression of TCF4 while monitoring transcript and protein levels. Plasmids for multiplexed ... new serotype PHP.V1 that exhibits efficient transduction of vesicular brain cells. Find this AAV at ...

...amplification) or reverse transcriptase (RT)-RPA, respectively. RPA is coupled with T7 transcription to convert amplified... method termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen et al., 2018). Both SHERLOCK...almost anywhere as lateral flow strips are easy to transport and work rapidly, providing reliable results in...    DETECTR- DNA Endonuclease Targeted CRISPR Trans Reporter Cas12a (Cpf1) is a CRISPR Cas variant that...discovered Cas12a’s ability to cleave non-specific (trans) ssDNA and harnessed this ability to create a DNA...DNase activity of Cas12a is initiated. Surrounding trans-ssDNA, including the ssDNA-FQ reporter are subsequently...Servellita V, Singh J, Miao X, Streithorst JA, Granados A, Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu ...

...promoter of another gene, thereby driving aberrant transcription. That being said, we generally pick guides...been significant work on constructing Cas9 transcriptional activators. These constructs allow for the ...Cas9 deficient in DNA cutting activity to a transcriptional activation domain (Fig 1). Two ways to use...picking whichever guides are closest to the transcription start site (TSS). We recommend targeting the...Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator Systems. PLoS ONE 11:e0158294 . https...Church GM (2015) Highly efficient Cas9-mediated transcriptional programming. Nat Methods 12:326–328 . https...Nishimasu H, Nureki O, Zhang F (2014) Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex...

... cell types. Specific transgene expression can be also achieved in nontransgenic animals by using cis ...delivery by increasing transgene cargo capacity, enhancing anterograde transport efficiency, and engineering...proteins that determine the AAV serotype. During transduction, the viral capsid proteins bind to the target...serotype used for a specific cell type. Find some transduction characteristics of select AAV serotypes in Table... AAV vectors are common approaches to control transgene expression by restricting expression to genetically...enhancers, multicistronic vectors, and post-translational regulatory elements. Examples of these approaches...

...enzyme or transcriptional activation, conformational changes, translation, and protein translocation. Targeting...triggers CaM/M13 interaction that increases FRET transfer between the fluorescent proteins. FRET is highly...characteristics like pH, redox state, and voltage across membranes. These sensors generally follow the same pattern...an organelle-specific targeting signal or a transmembrane domain, biosensors can be targeted to a particular...endoplasmic reticulum (ER), vacuole or cellular membranes. Examples include blue flame mitochondria-targeted...

... the CAR facilitates signal transduction and release of cytotoxic granules from the T cell. When a CAR...., 2012, Bates and Power., 2019): rapid blood clearance, which is useful for imaging applications; better...small size (Bates and Power., 2019). Their rapid clearance from blood can be a drawback for therapeutic applications... has led to their application for basic and translational research.  Basic research applications of scFvs...single molecule tracking in living cells. The HA frankenbody is an HA tag detection probe. It contains an ...to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based probe, but ...generate since it’s genetically encoded.   Translational applications of scFvs: Just like antibodies...

...ready-to-use viral preps   Improved cell-free transcription/translation system to examine biosynthetic pathways... Gabrielle Clouse Recently, cell-free transcription/translation systems (also referred to as TX-TL systems...modifications to edit DNA without requiring a double-stranded break. The Xiao Wang Lab has recently deposited.... These plasmids use “Cas9-mediated adenosine transient reporter for editing enrichment”, or XMAS-TREE...and enrichment use of XMAS-TREE. a) Cells are transfected with three different vectors and flow cytometry...identification of essential MYC binding sites in a wide range of human cells.  A new pooled library from Junjie...

...fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced, stochasticity...fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein level. The...gene delivery is more common than conventional transgenic strategies. In such vector-based gene delivery...in the original method, we used the Tetracycline trans-activator system to boost the expression levels,...levels, and we named the system Tetbow (Tetracycline trans-activator Brainbow) (Sakaguchi et al., 2018) (Figure...expression level of XFPs, most likely by suppressing transcription. It is critical to express a minimal amount ...Bio-Protocol 8 (2018): e3046. Livet, Jean, et al. "Transgenic strategies for combinatorial expression of fluorescent...

... and not in academia is run by grant funding. Learning to write grants quickly, efficiently and effectively...apply to or offer to help with lab grant writing. You may not get the grant or your PI may edit everything...thinking about future career options. 5. Write a grant - even if you don’t need one A lot of science in...valuable skill. There are many different kinds of grants that you can apply for. If you are in a lab, your...Here are some sites that have compiled lists of grants: Science: Where to Search for Funding The Ohio...For Your Research University of Florida: Training Grants and Other Funding Opportunities L’oreal USA For...

...grateful to the grant officers at Fast Grants. We received a miraculous grant from Fast Grants, which was ...studying the coronavirus. Addgene’s Technology Transfer Team also moved fast to ensure that the majority...collaborations as well as provides visibility for grant-driven research being done in all labs sharing materials...materials via Addgene. Perhaps if each of many grantors supporting research could provide Addgene with... for supporting Addgene by providing a generous grant. This support will enable us to continue all operations...

...method is transfection of short interfering RNA (siRNA). Knockdown caused by siRNA is also transient, and ...interest. This approach often involves the transient transfection of expression plasmids in a given cell ...commercially available. Various proteomic and transcriptomic datasets can be used to select a candidate ...antibody should be able to detect even in cells transfected with low efficacy. But since there can be large...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...workshops. Let’s use TIM-1 (Single pass type-1 membrane protein), which has been designated CD365 in the...the antibodies both recognized the target when transiently overexpressed in CHO cells and displayed similar...

...2015, there were six of us. We were initially strangers but we all shared a passion for DIY Biology and...December 2015 we held our first intro to bacterial transformation course (read our blog post on it and to see...consisting of over twenty members and volunteers. Transforming citizens into scientists Anyone can become a...dedicated time at BosLab, he became our bacterial transformation guru and instructor for our intro to biohacking...biohacking courses. Our members pursue a diverse range of projects at BosLab. Some are entrepreneurs using...RNA with small molecules. Other members’ projects range from hacking probiotics to filming educational lab...BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge with the community...

...covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR template. Some of the applications...by using NEBuilder which requires only a single-stranded synthetic oligo to generate a single-guide RNA...RNA (sgRNA). Using only one single-stranded oligo, instead of the more typical two oligos, is more cost...annealing step usually used to create a double-stranded oligo. Generating an sgRNA construct becomes as...target region. Assembled plasmids can be directly transformed into E.coli for plating, screening and generation...complexity, is achieved with synthetic oligos with a randomized region between the regions of homology. Producing..., generating expression cassettes for yeast transformation can take 5 days. NEBuilder shortens that time...

...differently. Communication comes in many forms—from transparency about data to allow others to see what you actually...communication and sharing fits together to make for a transparent and collaborative research environment. Journals...Currently, most of the burden for pushing for more transparency in science communication falls on the shoulders...developed STAR Methods to improve the structure and transparency of methods sections, including clear information...researchers are not strongly rewarded for being transparent in their communication or making their data and... to a large extent, hold the keys to getting transparency and communication further up the collective ...management plans and for evidence of sharing for grant renewal—but there is clearly room to do more. If...

...various teal, green, yellow, orange, and red fluorophores. The spectra for mOrange2 (scaffold of OCaMP) are...are shaded in orange. Data from FPbase.org. Scientists have developed calcium indicators spanning ...complement these tools, we recently developed a new orange calcium indicator, OCaMP (Aggarwal et al., 2025...2025), based on the fluorescent protein mOrange2 (Figure 2). Figure 2: OCaMP structure and mechanism...CC-BY-NC-ND 4.0 International license. Why orange? OCaMP’s emission peak at 565 nm fits neatly between...for two-photon imaging with newer lasers in the range of 1000–1080 nm. These industrial fiber lasers are...References Aggarwal, A, et al. (2025). A sensitive orange fluorescent calcium ion indicator for imaging neural...