...neuroscience, opsins are typically ion channels that allow charged particles to cross the cell membrane ... signal. The non-specific cation channel ChR2 and the calcium channel LiGluR both cause ion movement that...exposed to light. Neuronal firing depends on these charged particles and whether they make the inside of the...different opsins, studies have shown that some channels, such as ChR2 and NpHR, can even be expressed ... the escape response neural pathway is well characterized, there is much left to be discovered of spontaneous...

...undergoes a conformational change in response to the molecule it detects. This change then generates a reporter...sensing domain induces changes in a fluorescent signal based on ligand presence or changing cellular conditions... calcium induces CaM/RS20 interaction, thereby changing the FP’s structural organization and activating...Pertz’s lab. Other biosensors track cellular characteristics like pH, redox state, and voltage across membranes... or transcriptional activation, conformational changes, translation, and protein translocation. Targeting...

... fluorescent data on all channels - helpful if you forget to set the channel wavelength before you run... most consistent in your hands.  When using multichannels, set up your plate in a way that will tell you... the board. If the samples or sample conditions change, the multiplexed protocol may have to be re-optimized..., which gives an RQ value representing the fold change of each sample compared to your control. But as...Wong, 2018.    🔥Hot Tip🔥 Negative fold changes in ΔΔCT are represented by RQs between 0 and 1.... during analysis can allow you to see this fold change as a positive difference (RQ>1), which may be easier...s41598-017-02217-xTaylor, SC; Nadeau, K; Abbasi, M; Lachance, C; Nguyen, M; Fenrich, J. The Ultimate qPCR Experiment...

...they’re reading about weren’t changed halfway through in order to chase a positive result. Beyond these...address this issue directly by creating well characterised antibodies against all human extracellular proteins...play a role here by making it easier to retain searchable, thorough notes on how lab procedures were actually...similar experiments. Biology is messy enough that by chance, some small number of the scientists will get a... experiments on similar subjects in a single, searchable space. Finally there are new publishing outlets...of peer reviews so we can see how and why papers change throughout the review process. This will hopefully...problem in Button et al 2013 and Ioannidis 2005) and changing an experimental set-up partway through an experiment...

...a drug, including the product origin; purity, mechanism, and stability; and sequence of biologic product...Efforts to predict T cell epitopes focus on characterizing those epitopes as CD8 or CD4 epitopes, as well...of precision medicine in the future.   Madhura Acharya is a former co-op and current Scientist, DNA Sequencing...B., LeSauteur, L., & Burns-Naas, L. A. (2015). Chapter 27—Immunotoxicology Testing of Monoclonal Antibodies...Formation in Oncology: Clinical Relevance and Challenges. The oncologist, 21(10), 1260–1268. https://doi.org.../journal.pone.0159328Groell, F., Jordan, O., & Borchard, G. (2018). In vitro models for immunogenicity...and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V...

...three additional amino acid changes were introduced to nCas9. These changes promote nCas9’s dissociation...molecular biologists use to tinker with DNA. Making changes to a DNA sequence can help scientists identify ...components of EvolvR and its potential applications. Challenges of existing mutagenesis methods Several continuous...drawbacks with these approaches that makes it challenging for users to achieve targeted diversification...types of nucleotides. Both the Bassik Lab and the Chang Lab have created methods that introduce diversification...Cytidine Deaminase (AID) enzyme. AID, however, only changes cytidines or guanines to the other three bases,...

...articulating why the results you've measured should change established theory. So, what can you do to ensure...you receive generalizes.  If someone suggests a change to a slide, think about whether the same principle...explain why the work is important and how it should change how the members of your audience think or act.   ...of science, communication isn't easy.  But the challenge is worthwhile and rewarding. A truly passionate...comscicon18 and stay tuned to our website and social channels to apply for our future events. In addition to...comscicon.com/. All our events are offered free of charge to accepted students, and we pay for travel expenses...Nathan Sanders is a co-founder of ComSciCon and Chair of its Leadership Team.  Nathan did his PhD in astronomy...

...light chain sequence, but was predicted to occur at a low frequency within functional light chain sequences...IgG variable light (VL) and variable heavy (VH) chain sequences. Fusion PCR was performed to create an...backbone for expression of the mAb heavy and light chain, each from a constitutively active CMV promoter.... coexpression of light (green) and heavy (blue) chains as driven by two CMV promoters (orange). Figure...R-mAbs. It was recognized that an aberrant IgG light chain transcript was expressed by some of the hybridomas..., which in most cases left the functional light chain intact, the authors were able to greatly increase...

... can’t fix an experiment by simply running it unchanged over and over again, and the same goes for your...again (I’ve been there!), think of ways you could change your variables - your approach, your workflow, ...setup - to prevent the mistake, just like you would change variables to fix a failed experiment.  For example...work, and the possibilities for small, helpful changes are endless.    Fig. 2: Blugene moves ...intentionally slow down, you’re giving yourself a chance to break what can be a rather vicious cycle.  Everybody... are, however, many ways for you to reduce the chances of making an error. Try a variety of things, including... experimental approach, use small, incremental changes to understand what works and what doesn't. By being...

...fluorescent output, which will turn on, change color, or change intensity of fluorescence in the presence...receptor, but readouts could involve fluorescence, pH change, heat, etc. The last component of the biosensor...Calcium triggers muscle contraction, and calcium channels are activated for neuronal action potentials; ...actinin will produce different signals based on changes to the structure of actinin in low vs. high tension...real-time cellular reactions to stimuli and as well as changes in cellular dynamics during so-called housekeeping...Viral delivery has several advantages over other mechanisms. For one, many viral platforms, such as AAV or...

...That’s what makes it so wonderful. The most challenging part of jumping into this career path from academia...key in science policy. Luckily, there are many mechanisms in place to ensure your effective networking-...disease area. Depending on the organization, these changes to policy could shift how society interacts with...or essays. Note: the answer to this question may change over time and that’s perfectly normal!  Action ... your hair out, your successes, failures, and challenges you’ve overcome. This information may help you...what parts of your current job you want to keep or change and can inform your move into science policy. Read...completely new field without issue. This may be more challenging if you are trying to apply for an outright job...

... reinforced by the findings of the 1952 Hershey-Chase experiment. In the same year, Rosalind Franklin ...meganucleases, were discovered. These proteins are characterized by large DNA recognition sites, which makes ...makes them highly specific.  Jurassic Park by Michael Crichton was published in 1990, just a year after the...– A, T, G, and C. In this movie Ethan Hawke’s character revolts against class divide that is created by...protagonist (Langdon). The plot involves Langdon chasing an anti-villain geneticist named Zobrist who designs...CBE) use a cytidine deaminase to introduce a C→T change at precise locations in the genome. This was soon...doi.org/10.1534/genetics.111.131433 Doudna JA, Charpentier E (2014) The new frontier of genome engineering...

...Castle, Michael J., et al. "Adeno-associated virus serotypes 1, 8, and 9 share conserved mechanisms for anterograde...Astrocytes AAV8 > AAV2 & AAV6 Modified from Aschauer et al. (2013) What does this mean to us? Well,... (anterograde transport), which is a possible mechanism for the AAV to infect other cell types not within...transport, but they also have similar transport characteristics (velocities of movement, distance traveled,...16.6 (2008): 1073-1080. PubMed PMID: 18414476. 3. Aschauer, Dominik F., Sebastian Kreuz, and Simon Rumpel...PMC3381869. 7. McCarty, D. M., Paul E. Monahan, and Richard J. Samulski. "Self-complementary recombinant adeno-associated...Central PMCID: PMC190186. 9. Grieger, Joshua C., and Richard J. Samulski. "Packaging capacity of adeno-associated...

...unlike most of the other mechanisms described. Fig. 3 dTag mechanism of protein degradation. Adapted...be enough, or do you want a permanent control mechanism? Where is the control happening? Is the regulation...need help choosing. Viral expression is another mechanism to integrate your choice cDNA. Both lentivirus... Fig. 1 CRISPR activator and repressor mechanisms Want to transiently induce expression? Check... its analogs will induce these transcriptional changes, and expression is tunable by titration with different... are akin to degron tags, just with a flipped mechanism. When a protein is tagged with a DD, it is unstable...

...disease characterized by increased iron serum levels and early-onset cataracts, is characterized by point...want to study RNA-protein interactions? The characterization of RNA-protein interactions has lagged behind...bind an RNA motif of interest in living cells. Challenges with existing RNA-protein interaction methods...containing three well conserved but not well characterized RNA motifs. This bait identified a new interaction...Ramanathan et al. found evidence that suggests changes in RC2H1 protein expression and post-transcriptional...Response Element (IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE binding...

...doi.org/10.1038/nmeth.3312 Chavez A, Tuttle M, Pruitt BW, Ewen-Campen B, Chari R, Ter-Ovanesyan D, Haque...for each loci, CRISPR/Cas systems only require changing the programmable guide RNA. Gene activation by...Tanenbaum et al., 2014, Konermann et al., 2015, Chavez et al., 2015). Most popular methods for CRISPR...other popular activation methods (VPR and SunTag) (Chavez et al., 2016). SunTag Description Rather than using...method is its construction: it relies on antibody chains, which are relatively large and are not expressed...:7429–7437 . https://doi.org/10.1093/nar/gkt520 Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R...

... unpredicted gene expression changes (Sapozhnikov and Szyf 2021, Charlton et al. 2020). Most importantly... such as dCas9-TET, can, like the iron ball-and-chain that they resemble, actually impede your investigation... Figure 1: Break free from the ball and chain!   Epigenetics nation stuck with methylation...methylation state causes a condition. To understand mechanisms and, eventually, develop treatments, we need ...Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, Mattei AL et al. (2020) TETs compete... https://doi.org/10.1038/301089a0. Galonska C, Charlton J, Mattei AL et al (2018). Genome-wide tracking...

...stable cell line. The genes encode the heavy chain and light chain for the antibody and when translated into...Figure 2: (Left) Figure 2: (Left) The light chain and heavy chain genes of the antibody are encoded in a plasmid...the line and subsequent cloning of the antibody chains (Crosnier et al., 2010; Andrews et al., 2019). ...reagents. These include conventional heavy and light chain recombinant monoclonal antibodies as well as other...With molecularly defined antibodies that do not change over time, scientists will know exactly what antibody...

...systems tend to have slower, but larger, calcium changes. The GCaMP6 family of sensors are commonly used...compartments can vary greatly. Additionally, the calcium changes you will observe in your experiment could be subtle...rearrangements of the sensor, and (3) subsequent change in fluorescence only occurs at high calcium concentrations...are available to simultaneously measure calcium changes in the ER, cytosol and mitochondria), the Cameleon-based...ratiometric GECO sensor (GEX), where one wavelength changes with Ca while another wavelength is insensitive...sensors have also been re-engineered to measure changes in calcium specific to the neuronal cell body or... Shen Y, Kramer L, Campbell RE. Structure-and mechanism-guided design of single fluorescent protein-based...

.... For example, the light-gated cation channel, Channelrhodopsin 2 (ChR2), exists in several states: closed...of the patch cable (although you may be able to purchase one pre-made instead). Laser stimulation     ...      In the video above, Channelrhodopsin 2 (Chr2) has been expressed in the motor cortex of a mouse...determine the ideal excitation). Lasers can be purchased through Laserglow, which caters to optogenetics...PMID: 22157972. 3. Britt JP, et al. Use of channelrhodopsin for activation of CNS neurons. Current protocols...editorial board, Jacqueline N Crawley  [et al]. 2012;Chapter 2:Unit2 16. doi: 10.1002/0471142301.ns0216s58. ...