...biological tools, and with the right safety considerations, they aren't dangerous for controlled lab usage...and episomal expression will last ~2-7 days at moderate levels. With regard to target cells or organism...Transduce on! Resources Resources on Addgene.org Generating Stable Cell Lines with Lentivirus Virus Handling...

... due to a few limiting factors.  To start, the average health science startup requires a great deal more...those ideas into commercial products and next generation companies.  We launched in the early spring of...(CFO).  As CEO, I am responsible for internal operations and building external strategic partnerships ...

...enter cultured cells and activate expression of a luciferase reporter. CPP was not required for protein entry...cells! After verifying the specificity of their luciferase assay, they concluded that components in the ...lethality and were sorted into single wells to generate clonal populations. After iTOP treatment, both...

...However, careers outside of the lab highlight “transferable” skills such as leadership, management, and ...impact your work? What design variables need consideration? Enrich your knowledge and skills outside of...tools such as the Individual Development Plan to generate a list of potential professions Google potential...

... with a simple question: how do Cas9 and sgRNA interact with and dissociate from a DNA target? The surprising...et al., 2016. We next wondered if we could leverage this model (built from in vitro studies) to boost.... Figure 2: Single-stranded donor DNA interaction with the Cas9-DNA duplex. Image from Richardson...

...reading, you can calculate the DNA concentration. Generally, A260 of 1.0 is equivalent to 50 ug/ml pure dsDNA...samples and are often used to quantify DNA for next generation sequencing. Unlike absorbance-based methods, .... This method is often used preceding next-generation sequencing or microarray studies and is not used...

...minor variation in experimental conditions will derail a research program (think a subtle difference in...to help your fellow researchers advance science overall. Communication is the issue Much of the problem...President. Her current responsibilities encompass overall leadership of the Cell Press editorial department...

...example, the hedgehog gene can be overexpressed in several cell types by using different Gal4 drivers. It ...explain the relatively straightforward process of generating your own fly lines. In the meantime, be sure ...expression as a means of altering cell fates and generating dominant phenotypes., Development 118, 401–15...

...-rings are, in fact, permeable to CO2 at low temperatures (-80°C) and once in the tube, the CO2 can alter... storage, we then thawed the samples at room temperature and used the color of the liquid solution and...not all O-rings) were permeable to CO2 at low temperatures. We tested two different types of tubes: the...

...in a cell or animal model. AAV plasmids contain several important elements, but among the most important...occurring.  How do I validate my ITR sequences?  Next-generation sequencing and Sanger sequencing might fail to...vector stability and sequencing results. Next-generation sequencing is also helpful in quantifying ITR...

... need a second GFP antibody?  Similarities:  Generated against full length GFP and converted to recombinant... also withstands denaturing conditions, high temperatures, and other chaotropic conditions better than...fluorescence in high molarity GdnHCl, at high temperatures, and in the presence of oxidizing agents. Adapted...

..., why it affects our emotions, and even what therapeutic properties it has. As with other works of Dr....of characters from five different families over several millennia as their paths cross with each other ...Nicholson Baker’s The Anthologist. The book ended up literally changing my life. Though I have an undergraduate...

...landmark Cell paper showing that it was possible to generate induced pluripotent stem cells (iPSCs) capable...has spent years studying how that one protein interacts with that one promoter can tell you, it can be...clinical trial patients with advanced macular degeneration; Laurenz Studer discussed his work with Parkinson's...

...out of these interactions, ask yourself how every piece of feedback you receive generalizes.  If someone...skillset rather than a permanent fix. Treat every interaction in your work as an opportunity to practice communicating...

... how stretches of DNA on different chromosomes interact with each other. Chromatin regulation is complex...in the right direction! CAPTURE-ing Chromatin Interactions: Using CRISPR-dCas9 to Study Gene Regulation... An Improved Method for Studying Protein-DNA Interactions CRISPRainbow and Genome Visualization CRISPR...

...Transfection is the process by which foreign DNA is deliberately introduced into a eukaryotic cell through non-viral...diethylaminoehtyl (DEAE)-dextra neutralize or even impart an overall positive charge on DNA molecules so that it can.... "Chitin induces natural competence in Vibrio cholerae." Science 310.5755 (2005): 1824-1827. PubMed PMID...

...talk at the Cell Symposia: Gene- and Cell-based Therapies: CRISPR, Stem Cells, and Beyond meeting earlier...III. Gene III is under the control of a T7 RNA polymerase and cannot be expressed until the base editor...to an uninfected culture which results in less overall dilution and less stringent phage selection. ...

...and UAS plasmids at Addgene Choosing a vector to generate a transgenic fly There are a few important points... when choosing the vector you’d like to use to generate your new fly line: 1. Where would you like the...  et al. Cytokine/Jak/Stat signaling mediates regeneration and homeostasis in the Drosophila midgut. Cell...

... the “Analyze Sequence” button to pull up an interactive plasmid map powered by SnapGene and gain access...the static maps discussed above, but also have interactive functionalities. You can now hover over any feature...maps, the linear sequences hosted here are now interactive and have similar hover over features. As with...

...protein is based on the properties found in the literature or tabulated data that summarizes properties ...). However, the theoretical brightness ignores several important experimental conditions that are related...detector sensitivity) and the biological system (temperature, prokaryote versus eukaryote, background fluorescence...

...closely associated and function as one unit: Gβγ. In general, GPCRs show a propensity to associate with certain...receptor, which can then activate expression of a luciferase reporter in the nucleus. The Roth lab showed ...Chemogenetics uses genetically engineered receptors to interact with small molecules to elicit a response in a...

...Plasmids 101 will provide you with an overview of general molecular biology knowledge and techniques. If ...with only white colonies is very suspicious! Refrigerate your plates: Placing plates at 4C for few hours...care in making your plates: X-gal is light and temperature sensitive and needs to be added to media after...

... of this is gospel, but we think it's worth consideration. Everything will go wrong Whenever an order ... lab, a similar situation could arise if a lab veteran who worked with a particular sample suddenly left...easily as needed. In this case, the physical considerations of the laboratory make object composition a...

...exposed to 500 nm violet light, the Dronpa dimers interact and lock MEK1 in an “off” setting by blocking ...possible to create other photoswitchable kinases? The general design of psMEK1 also worked for creating three...all-optical psMEK1 + ERK KTR-mRuby2 system has several advantages over existing methods. First, it avoids...

...the photostability measurements reported in the literature are performed in different ways. We describe ...intensity remains. The first issue is that, in several cases, the fluorescence intensity decay of a fluorophore... be identified by a head-to-head comparison of several fluorescent proteins of the same spectral class...

...implementing consensus recommendations, even when a federal mandate encourages them to do so. Immediately after...There are institutions in Boston - Massachusetts General Hospital, Mass Eye and Ear Infirmary and Beth Israel...salaries when all others in Boston (some of them literally across the street from these institutions) are...

...CRISPR-Cas9 and a custom gRNA is all you need to generate a frameshift mutation that will permanently ablate...package cDNAs for genomic delivery and have many therapeutic advantages, such as tissue-specific targeting... to temporarily increase expression. There are several flavors of activators which can be used with this...

...from peer-reviewed publishing in a journal.  Accelerating Scientific Communication Preprints have been...Fig. 1: Stand aside, CERN - it's a new age of accelerating science! Photo courtesy of under a CC license...attendees to read and comment online! Want the general public (say, policymakers or community organizations...

... randomly. Therefore, inserting loxP sites at deliberate locations in a DNA sequence allows for very specific...reciprocal crossover event. This event can have three general outcomes based on the location and orientation ...separate DNA molecules, a translocation event is generated at the loxP sites. How can I use Cre-lox? The ...

...and think of ways to relate the proposal and deliverables to the aims requested in the call. Step 2: ...technologies. Therefore, take time to review the literature and offer educated insight into what the desirable...each aim. A funded proposal typically lists the deliverables, explains 1-2 potential pitfalls of the intended...