... norvegicus genes! Mexican axolotl (Ambystoma mexicanum) Ambystoma mexicanum, aka the axolotl. Image...essential aspect of biological research that has defined our understanding of biological processes and developments...studying genetic modification for their ease of manipulation and their ability to be replicated. Because E....providing insight into some of the most significant and fundamental biological processes. Many of their genetics...tools in developmental and biomedical research.  Xenopus, aka African frog. Image from Holger Krisp...Advances on Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306...Organism. News-Medical. Retrieved on September 18, 2020 from https://www.news-medical.net/life-sciences...

...the grant application process. When my PhD advisor first asked me to write a grant application due the ...pursue. Proposal writing: Improves technical writing and communication skills, Elucidates the big picture...information required for a successful grant application. It typically includes overview information, key dates...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...Research at universities typically requires funding from a variety of government and academic institutions...advisor alone applies for these competitive grant applications; however, in some cases, your advisor may choose...

...Other than a theoretical maximum value, physical titers don’t necessarily give any indication of the infectious... red) report physical titer, which is measured by qPCR.   Physical titer is typically calculated by... you? Physical titer: the concentration of viral particles containing viral genomes. Physical titers ...qPCR or other DNA quantification methods - see below), since each viral particle typically contains one viral...facilities report the physical titer of a viral solution (Figure 2). Because physical titers are used for...titer values. Finally, because physical titer does not give an indication of infectious titer, consider...that can transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type...

... of the American Chemical Society 123.48 (2001): 12123-12125. Journal of the American Chemical Society...discovery–a review of theoretical aspects and recent applications." Current Chemical Genomics and Translational...originally developed to treat heavy metal intoxication in chemical warfare [Vilensky & Redman, 2003]). Most...al. "Designing logical codon reassignment–Expanding the chemistry in biology." Chemical Science 6.1 (2015... of a biarsenical− tetracysteine motif determined by NMR spectroscopy." Journal of the American Chemical...laboratory, then FRET experiments are a viable biophysical path to answers. What is FRET and what's it...Fluorescence resonance energy transfer (FRET) is a biophysical interaction between two fluorophores. When the...

...underlying all communications efforts.  Scicomm definitions (as used by Addgene)    Science communication, or science...time engaging with science communication efforts.  Scientific communication or scientific writing is peer...to note that it is the actual communication itself, not the communication method, that determines if something...scientific or science communication. A Tweet is still considered scientific communication if the language is...Series, where we’ll be talking about science communication, best practices in the field, and even getting...getting into some active science communication research.   This summer, we’re making time to talk about ...about my favorite thing - science communication (scicomm)! We have so many fun topics planned for this summer...

...epigenome (the sum of epigenetic modifications across the genome) has been implicated in disease states, and targeting...DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of epigenetic...Preps for Epigenetic Modification Why use epigenetic modifiers? Epigenetic modification is certainly not ...Nature Communications 8: . https://doi.org/10.1038/ncomms16026 Find plasmids from this publication at Addgene..., 2017 and updated Jun 24, 2020. Epigenetic modifications are an additional layer of control over gene...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...mechanisms remain unclear. Today, targeted epigenetic modification at specific genomic loci is possible using CRISPR...

...or DYKDDDK 1 Affinity and Purification Good for antibody-based purification; has inherent enterokinase...site GST Large Protein 26 Purification and Stability Good for purification with glutathione; protects...used for purification as elution requires low pH poly His HHHHHH  0.8 Affinity and Purification Very small...GKPIPNPLLGLDST   1.4 Affinity and Purification Good for antibody-based purification Combo and cleavage tags...increasingly common. Tandem affinity purification (TAP) once referred specifically to a combo tag comprised of ...multitude of uses including (but not limited to) purification, detection, solubilization, localization, or...overexpress and purify proteins can face any number of technical challenges depending on their protein of interest...

...differing in pigmentation." Biochemical and biophysical research communications 78.1 (1977): 237-243. PubMed...one that can be achieved using pharmacological or electrophysiological methods (Deisseroth, 2011; Mei ...recombination. Purification and properties of the Cre recombinase protein." Journal of Biological Chemistry...Edward S., et al. "Millisecond-timescale, genetically targeted optical control of neural activity." Nature ..., et al. "Next-generation optical technologies for illuminating genetically targeted brain circuits." ...and Jinfang Zhu. "Immunologic applications of conditional gene modification technology in the mouse." Current... R. Thompson. "Overview on research and clinical applications of optogenetics." Current protocols in pharmacology...

... construct for every application. Plus, the photophysical properties of chemical dyes provide many advantages...Few Complicated Non-natural amino acids 1 Various Many Complicated   Applications Organic... W. (2011). Chemical Tags for Labeling Proteins Inside Living Cells. Accounts of Chemical Research, 44...If you take a look in a typical lab freezer, you’ll probably find lots of different plasmids containing...new green fluorescent protein, some affinity purification tags, another version with that same red fluorescent...substrate. If the substrate was linked to, say, a chemical dye or other functional ligand, your protein of...advantages over fluorescent proteins, opening up new applications.   Figure 1: Live-cell labeling strategies...

...Spatially Confined Persistent Diradical Emitter. Journal of the American Chemical Society, 145(25), 13615–13622... tools for familiar applications. But how does one invent a tool for applications that don’t even exist...organisms) are opaque, which makes most biomedical applications of these tools difficult or impossible....of developing new microscopy tools for biological applications across a wide range of topics. He embraces...., Timmel, C. R., & Hore, P. J. (2016). Chemical amplification of magnetic field effects relevant to avian...phenomenon, improve the tool, and develop new applications for magnetoresponsive proteins. Motivated by...tool with magnets would enable new research and applications that don’t yet exist, but it wasn’t clear that...

...of Various Motifs is Critical for Initiation of Replication. Journal of Biological Chemistry, 273(14), ...to utilize diverse host replication machinery. For instance, the ori typically contains iterons that serve...ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various hosts...the ori and allow for replication. However, if plasmids have their own replication machinery such as Rep...different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids have a narrow... while others have a broad host range and can replicate and stably maintain the genes they carry among.... These features are: The plasmid origin of replication (ori) contains structural elements that are versatile...

... your own research! Classification Structure Principles of CRISPR classification While CRISPR systems... CRISPR-Cas systems. Philosophical Transactions of the Royal Society B Biological Sciences, 374(1772),...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...technologies. Learning the general principles of this classification system will help you gain a deeper appreciation...multi-pronged analysis to sort effectively. The current classification system relies on a combination of sequence ...a similar function. After analysis, CRISPR classifications were placed into a fairly simple hierarchy:...

...interaction between a bisarsenical dye ligand (e.g., FlAsH, ReAsH) and a short genetically encoded tetracysteine...systems consist of a genetically encoded enzyme variant tag that reacts specifically and irreversibly with... as intact tissue. Some chemical labeling strategies such as the bisarsenical dye (Figure 1a) and FAP ...Ronald T. Raines. "Bright ideas for chemical biology."ACS chemical biology 3.3 (2008): 142-155. PubMed... at the Janelia Research Campus, Howard Hughes Medical Institute. Chemistry is dead, long live chemistry...fluorescent protein (GFP) sparked a renaissance in biological imaging. Suddenly, cell biologists were no longer...synthetic fluorophores. Add a dash of DNA with an electrical jolt and cells become perfectly capable of synthesizing...

...Retroviruses are not considered safe for clinical applications.     Lentiviral: Lentiviral vectors infect...lentiviral delivery is not considered safe for clinical applications. Excisable Cre-lox lentiviral vectors:... to reprogram. Basically, there’s no universal method that can handle all applications of iPSCs, but there...generation of iPSCs is relatively simple in concept: ectopically express a cocktail of stem cell reprogramming... to verify there’s no genomic integration Replicating EBNA1 episome Non-integrating Medium Medium ...creates heterogeneous iPSC cell lines, which can complicate comparisons made between lines. Incomplete silencing...unavoidable heterogeneity between clones. This can complicate comparisons made between samples, i.e. transgene...

... (enChIP) using CRISPR." Biochemical and biophysical research communications 439.1 (2013): 132-136. PubMed...affinity purification with an anti-Cas9 antibody. These different tag systems and affinity purification schemes...26848818. Fujita, Toshitsugu, et al. "Identification of physical interactions between genomic regions ...genomic alterations. These studies utilize a catalytically inactive form of Cas9 known as dCas9 (Jinek ...interest targeted by a gRNA. CRISPR-mediated purification of a specific genomic region In 2013, the Hodaka...Subsequent mass spectroscopy studies allowed for the identification of proteins associated with the IRF-1 locus...steps (Fujita et al., 2013). CRISPR-mediated purification of a specific genomic region alleviates these...

...transfected (see Technical Tips). Minigenome replication and transcription In order to replicate the minigenome...transcription and replication strategies of Marburg virus and Ebola virus by using artificial replication systems...allowing researchers to study aspects of the EBOV replication cycle under BSL-2 conditions (4). What is a ...that are crucial cis-acting signals for genome replication and viral gene transcription (6). The EBOV genome...5’ ends of the genome which are required for replication and transcription (Figure 1). All viral genes...cis-acting signals necessary to direct minigenome replication and reporter gene transcription by the EBOV polymerase... in 2, 5). The minigenome is capable of being replicated and transcribed in the cell when the appropriate...

...resistance-mediating polymorphisms in African samples of Plasmodium falciparum." Journal of clinical microbiology 51.8 (...furiosus." Acta Crystallographica Section F: Structural Biology and Crystallization Communications 61.12 (2005... In vitro, ligases (notably T4 DNA ligase) are critical reagents for many molecular biology protocols,...or DNA base pair mismatches (8-11). For some applications, there cannot be any end-joining activity, and...regions. Why use a high fidelity ligase? Numerous applications have been developed to take advantage of the...Taq and other thermostable DNA ligases. These applications detect specific nucleotide sequences with high...rounds of melting and annealing allows linear amplification of the probe ligation product. With the right...

...D) teams at pharma companies carry out preclinical and clinical research to discover and test the best...years. Complicating these challenges is the fact that they occur in the context of significant clinical...training in reviewing literature, and critical analyses skills were applicable to my cases. I can directly use...molecular behaviors of chemicals and proteins to learn how a particular drug or medical device works. This... consulting? Healthcare consultants advise pharmaceutical companies, biotech start-ups, health insurance...efficient ways of delivering healthcare. Clients typically engage consultants as short-term experts to work...acquisitions of or collaborations with other companies. Medical Affairs groups work on building awareness among...

... receptors which play a critical role in development and are often implicated in disease progression. ...inability to replicate the spatio-temporal precision with which signaling events occur in a physiological setting...brighter mutant S65T-GFP, and the subsequent biochemical characterization, has enabled scientists to engineer...observations: Rapamycin is a membrane-permeable pharmacological agent that can rapidly induction heterodimerization...flexible cloning process provides improvements to classical cloning methods, particularly for complex transgenesis...enabled the group to perform independent two-color optical excitation of neurons. The paper describes five...setting. Genetic techniques typically rely on overexpression or knock-down, where signaling dynamics are...

...rather than being geographically constrained to the brain. In cases where it’s critical to minimize or eliminate...recommended for all systemic capsid applications. For example, AAV-PHP.eB is typically administered at a total dose...your specific application and animal model.  Other model organisms Tropism and efficacy can vary depending... 2023 * indicates viral preps available. Mechanisms of crossing the BBB are indicated in parentheses...crossing into the brain. While helpful from a biological standpoint, it makes delivering AAVs to the brain...throughout the organism to target large or diffuse biological structures. Of primary interest here are systemic...suitability of a systemic capsid for an individual application is dependent on a number of factors, including...