...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...

...“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...

...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...

...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...

... enhanced sodium accumulation in their roots and (3) development of tools for data analysis/visualization... to help decide whether work is suitable for publication in their journal. The early peer-review attempts...information that would prevent the study to be replicated or omission from the literature. I think that...Addgene.org Learn about depositing plasmids pre-publication Find molecular biology protocols Find eBooks ...

...2015 Dec 31. pii: aad5725. PubMed PMID: 26721683. 3. Nelson CE, Hakim CH, Ousterout DG, Thakore PI, Moreb... some research in human embryos. Nature. 2015 Dec 3. doi:10.1038/nature.2015.18947 5. Siva K, Covello ... by zinc finger nucleases. Mol Ther. 2015a Mar;23(3):523-32. PubMed PMID: 25492562. PubMed Central PMCID...setting. CRISPR is often used for animal germline modification, to repair or add in disease-causing mutations...that AAV-CRISPR does not cause obvious germline modification, an important finding given the controversy ...Blog Learn How CRISPR Could Be Used to Treat Cataracts and Cystic Fibrosis Read How Homology Directed...

...drug clinical trial, for example).   Fig. 3: Question flowchart for choosing an ANOVA/MANOVA statistical...positive is much higher than if you only look at 2 or 3 pairwise comparisons. Similar to family-wise error...for the nonstatistician: Part I. South Med J., 105(3): 126-130 (2012). 10.1097/SMJ.0b013e3182498ad5 Curran-Everett...tests  Statistical tests fall into two general categories: parametric and non-parametric. In this blog ... exist somewhere within your data; it does not indicate which groups the differences exist between. For...that matter. However, this is oftentimes an oversimplification. If your independent variables are affecting...

...conventional gene targeting is lowered to about 2-3 months. But certain models remain more difficult to...into a zygote and obtain a knockout mouse in about 3 months. Image courtesy of Wenning Qin. Single-...cleavage right before the stop codon!  Figure 3: How to design a C-terminal insertion with Easi-CRISPR... allele To design a knock-in allele at a given location, you’ll need to design only a single guide and... out our CRISPR Guide Find Plasmids for Cas9 Purification Find CRISPR Plasmids for Mammalian Expression...

...for 1-3 days. There’s also a CRISPR for that! CRISPR interference (CRISPRi) targets a catalytically dead...most of the other mechanisms described. Fig. 3 dTag mechanism of protein degradation. Adapted from...

... previously funded grant application to see how funds were allocated. Step 3: Get feedback from your ...proposal successful. Typical proposals consist of 3 to 5 aims, with secondary milestones to meet along...the grant application process. When my PhD advisor first asked me to write a grant application due the ...the next round of applications. Step 5: Keep writing! Each written grant application builds upon the ... project grant application" by the NIH Still want more information on the application process? What types...advisor alone applies for these competitive grant applications; however, in some cases, your advisor may choose...writing, volunteered to take the lead on various applications throughout my graduate career, and helped receive...

...PMC4364232. Find the plasmids from this publication at Addgene. 3. Ma, Hanhui, et al. “Multiplexed labeling...produce yellow, cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created... every genomic locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot induce gene editing...Cas9 ortholog, and the target sequence must be located adjacent to that PAM. To scale up fluorescent CRISPR...this rainbow of colors, Ma et al. performed sophisticated tracking of multiple chromosomal loci in live... do not induce genome editing. If used with catalytically active Cas9, the short, CRISPRainbow gRNA will... PMCID: PMC3918502. Find plasmids from this publication at Addgene. 2. Ma, Hanhui, et al. “Multicolor...

...shown below, a fragment with 5’ overhang TGGA and 3’ overhang TCCG can be ligated into a vector containing...to specify assembly order, so fragments with 5’ or 3’ sequence homology cannot be assembled using this ... work around this is to "domesticate" your fragment: PCR-based amplification can be used to create silent...and exciting applications, this system is an incredibly powerful tool for cloning complicated constructs...Type IIS sites (such as BsaI or BbsI), that are located on the outside of the cleavage site. As a result...plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning...restriction sites that may not be amenable to "domestication", you might want to consider using an alternative...

...genetically targetable fluorescent indicators. Nature protocols. 2006 Aug;1(3):1057-65. Resources on Addgene.org...(2) allosteric rearrangements of the sensor, and (3) subsequent change in fluorescence only occurs at ... a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium...in living cells and organisms. Unlike chemical indicators, these DNA constructs can be delivered to intact...the sensors discussed below fall into this GECI category.   For calcium sensors, one size doesn...:509-18. Inoue M. Genetically encoded calcium indicators to probe complex brain circuit dynamics in vivo...Kanemaru K, Iino M. Genetically encoded fluorescent indicators for organellar calcium imaging. Biophysical journal...

...like The Atlantic. We’ve been in documentaries in 3 languages (Japanese, German, and English) and even...we need to do it quickly. Hiring is hard but as a 3 year Boston Globe and BBJ Best Place to Work with ...enough space! There’s enough space here at our new location to grow. We are hiring again and getting ready...to keep up with the demand with excellence and dedication. We don’t have worry or wonder if providing...research, hosting conferences, and are expanding our catalog of materials to better serve the community. We ...projects allows us to support more new projects. Diversification is happening. The Addgene team will have some...we might do. I am grateful every day for their dedication, flexibility, innovative thinking, and openness...

...pulse of 500 nm light (turn on psRaf1) followed by 3 seconds of 400 nm light (turn off psRaf1). After this...is responsible for this MEK1/2 negative feedback. 3. Studying signaling pathways in vivo Expressing photoswitchable...members of their pathways, but where the kinases are located, their signal strength, and how long their signals...and MEK1 is active or “on.” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when...domains were inserted at sites homologous to the locations they were attached to in MEK. The photoswitchable...kinases good for? There are a number of potential applications for photoswitchable kinases.  Below are a few...signaling by using psMEK1 and an ERK kinase translocation reporter with a mRuby2 fluorescent reporter ...

...place before DNA replication. Indeed, they observed that 41/42 embryos were non-mosaic at 3 days post fertilization...showed a 3% mosaic rate, and this rate would need to be much, much lower before clinical applications could... embryos carried indels at the mutation site, indicating that repair proceeded through NHEJ rather than...attributable to CRISPR editing. Conclusions & implications One striking feature of this paper is the failure...experiments are needed before we can consider clinical applications for germline editing. Although this paper represents...