...-lantern emitted yellow-green light and exhibited 3-5 fold increased brightness over previous BRET probes... single-cell and whole-body imaging.” Nat Commun. 3 (2012): 1262. PMID: 23232392 PMCID: PMC3535334 Find...resulting in a fluorescent signal. Since their first publication in 2012, the Nagai laboratory has assembled a...multicolored nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion...sensitivity and faster imaging compared to previous publications. They also developed calcium, cAMP, and ATP ...furimazine. In addition to the previously described applications, the enhanced signal of eNL allows their use...characterizing Nano-lanterns and eNL for many different applications. For long-term imaging, especially where photobleaching...

...stable gRNA than when they inserted the aptamer at the 3’ end of the gRNA. Because of the success of the MS2...can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius. Triple color detection With...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without...has been widely adapted for a wide variety of applications: base editing, CRISPR activation and inhibition... (B) CRISPR-Sirius used to visualize relative locations of the subtelomeric region of the pericentromeric...al. “CRISPR-Sirius: RNA scaffolds for signal amplification in genome imaging.” Nature Methods 15 (2018)...

...Knife, Genesis 34, 1–15 (2002).PubMed PMID: 12324939. 3. S. E. McGuire, G. Roman, R. L. Davis, Gene expression...Versatile Toolkit for Gene Expression in Drosophila, 3, 1–10 (2015). PubMed PMID: 21356876. 5. J. Chow, Z...as overexpression of each gene individually. Applications of the Gal4/UAS system There are a number of...lines. In the meantime, be sure to browse the publication references and web links from my first post and...

...positive for your marker of interest (Fig. 3).   Figure 3: Isotype flow plot example. The isotype ...ll need a few controls specific to flow as an application. These controls will allow you to distinguish...commercial antibody pages. Flow cytometry can be a complicated and daunting process. Controls like single color...

...chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment (4-chloro-3-brom-...insert. All colonies on this plate should be blue, indicating that your IPTG and x-gal are working as they ...of false positives: Blue-white screening only indicates the presence of AN insert, not necessarily YOUR...

...-C CRISPR-Cas3 for genome-editing applications. Molecular Cell, 84(3), 463-475.e5. https://doi.org/10.1016... technology and applications with the type I CRISPR system. Gene and Genome Editing, 3–4, 100013. https...repression, high precision edits, and more. Figure 3: Schematic of the basic CRISPR-Cas9 mechanism, representing...double-stranded breaks caused by Cas12 create short 3’ overhangs, which leads to slightly increased efficiency... your own research! Classification Structure Principles of CRISPR classification While CRISPR systems...Shakespeare for the first time. The discovery and application of CRISPR-Cas9 kicked off over a decade of frenzied...properties. This led to a need to develop a classification system for the array of CRISPR technologies...

...resulted in a roughly 6-fold increase in expression [3, Figure 2]. Note how expression increases exponentially...PMID: 18500252. PubMed Central PMCID: PMC2574934. 3. Zingg, Brian, et al. "AAV-mediated anterograde transsynaptic... Protocols for Work with Virus Watch Our AAV Purification Video ...

...some impressive videos of these interactions (Ref 3). You can find useful tips on in vivo imaging in our...Research 9.12 (2016): 895-905. PubMed PMID: 27913448. 3. Cai, En, et al. "Visualizing dynamic microvillar ...Immunosurveillance and Defense With increasingly sophisticated visualization techniques it is possible to follow...

...no addition of nucleotides to the 5’ spacers. The 3’ ends were modified slightly with either a 1-7 base...constructs, transcript levels of PTG1 and PTG2 were 3 and 31 times higher, respectively. In rice protoplasts...biological pathways;  to observe a phenotype, the modification of multiple genes is often necessary. Guide ...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end. PTG construct schematics. Scissors indicate cleavage sites. After confirming gRNA cleavage...) How many loci can Cas9 modify? To test the application of this system to gRNA multiplexing, Xie et al...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct...

... cover letter can be found on the Addgene blog.   3. Personal statement brilliance In addition to sending... Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch for...your application a whole week before the deadline. The day of submission, review your application with... their dream.  To help students focus on the application process, and simplify access to academic resources...Scrutinize every opportunity prior to starting an application to assess your eligibility, interest, and career... detailed. Beyond the resume, a cover letter communicates your interest in a specific program, and institution...February represent peak season for internship applications. Thus these noble individuals are inundated ...

...Circuits through the Parafascicular Nucleus. Neuron 102(3):636-652. PubMed (Link opens in a new window) Marcinkiewcz...inhibitory opsins , genetically-encoded calcium indicators, and rabies targeting genes. Recombinases Addgene...

... the pH change caused by storage on dry ice (Fig. 3). However, 30 seconds was not sufficient to prevent... was a substantial reduction in infectivity after 3 weeks of exposure to pH 5.1 (Potter et al., 2014)....improve our shipment processes. After reading a publication describing how short term storage on dry ice ...medium, which is buffered and contains the pH indicator phenol red. We froze samples in a -80°C freezer...closed dry ice storage container for 48 hours (to replicate how our viruses are shipped), where they would...exposed to CO2/dry ice. All samples were tested in duplicate. What we found Not all tubes (or rather, not ... the samples were at the control pH of 7.4 as indicated by color (data not shown). What we did next: Venting...

...random-walk" of a budding startup, I asked Mike to outline 3 pieces that are essential to the biotech startup puzzle... anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes...competitions. After much learning, the focus of the application was changed, but Sample6 still leverages the ...fit for this set of technologies, effectively communicating the advantages embodied, and driving the enterprise...He explained that entrepreneurs must clearly communicate their idea at various levels of complexity and...backgrounds how amazing their idea is. While communication styles in academia and industry vary drastically...biggest challenge during this time was learning to communicate effectively to non-scientific audiences. "There...

...modified from pSC101 with copy numbers ranging from 3-100 per cell, a stabilized promoter buffered against... expression levels returned to the baseline after 3 hours with the stabilized promoter while gene expression...rounds of replication are initiated before the cell has divided. This means that genes located near the...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes...genes closer to the origin of replication will be overrepresented compared to those further away. As a...maximal gene expression near genomic origins of replication, the stabilized promoters created by Segall-Shapiro...the control of IPTG. Because RepA is a plasmid replication initiator protein, they could alter copy number...

... natural context or 3) be modified to extend at different pH levels. Domesticating R bodies We have shown...Protistology 48, 290–296 (2012). PubMed PMID: 22356923. 3. Polka, J. K. & Silver, P. A. A Tunable Protein Piston...actuators that could be used in a variety of applications. For example, you could use them to lyse cells...matrices) in response to pH changes. Beyond these applications, purified R bodies could also perform as actuators...PubMed PMID: 26814170. Find Plasmids from This Publication at Addgene 4. Petroni, G., Spring, S., Schleifer...

...provide problem-solving platforms.     Fig. 3: Different workflows involved in sharing open hardware... (2022). https://doi.org/10.1038/s41592-022-01602-3...convinced that most microscopy techniques can be replicated with commercially available consumer-grade components... tools has allowed the community to regularly replicate designs worldwide. Distributing hardware via globally...Hohlbein, et. al., 2022). A more open way of communicating in science brings stronger networking across...

...“blind” to this very important difference (Figure 3). If you are interested in harnessing the power ... PubMed PMID: 27089539. PubMed PMCID: PMC4835065. 3. Mock, U., Hauber, I. & Fehse, B. Digital PCR to assess...a gene of interest is one of the most common applications of precision genome editing, the remainder of...single molecule resolution of digital PCR allows classification of individual target molecules based on their...single wells of a 96-well plate without prior quantification. Comparatively, mismatch assays require between...

...recordings in primary visual cortex (V1) layer 2/3 (L2/3). B) Fluorescence from axonal boutons of neurons...Browse the IPI Glypican Collection here! Figure 3: General glypican domain structure. Glypicans are ...Here's what you'll find in this post: Glutamate indicators with increased sensitivity and tailored deactivation...viral preps targeting interneurons Glutamate indicators with increased sensitivity and tailored deactivation...popular choice for a variety of synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A...., Iyer, R., … Podgorski, K. (2025). Glutamate indicators with increased sensitivity and tailored deactivation...DNA vector that contains everything needed to replicate inside a host cell but with space to insert your...

...workflow. B) Detail of spacer peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm single alpha helix) and target...optimized PASSIGE plasmids here!     Figure 3: Schematic of the PASSIGE strategy. First, twin prime.... When the prime editor releases the DNA, the two 3’ flaps containing the edited sequence anneal with ...beads through an assembly of biotinylated SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP... (Figure 1). This enrichment streamlines the purification and minimizes sample loss while also supporting...scFvs for other targets or could be used for applications beyond cryo-EM.   Find plasmids for MagIC-Cryo-EM...interact with the air-water interface (AWI) before vitrification, they can be completely denaturated (left) or...

... near-infrared photosensitizer. Nature Methods,13(3), 263-268. doi:10.1038/nmeth.3735. PubMed PMID: 26808669...PMID: 18706812. PubMed Central PMCID: PMC4445427. 3. Bulina, M. E., Chudakov, D. M., Britanova, O. V.,...generation when bound by the dL5** FAP.  The iodine-modifications also shift MG-2I’s excitation wavelength to ...FAP-TAP system works. Find the Plasmids from the Publication Here Light-induced cell ablation with MG-2I-dL5...