...might think that being able to figure out what went wrong with a PCR is a pretty niche skill; outside of a...about seven different skills to you, you are not wrong. Problem-solving overlaps heavily with other skills...core part of our academic training.  Developing strong problem solving skills is just a matter of applying...As such, scientists are uniquely prepared to be strong problem solvers both in and out of the lab. The...position. Of course, there are limits to how far strong problem solving skills alone can take you - just...effectively does not mean I am qualified to be an astronaut. But, knowing that this skillset is a part of ... Ashley Waldron...

...be used for sparse labeling of neurons for tracing experiments.  Neuronal tracing experiments: While synthetic...broad number of cell types, such as inhibitory neurons, rods, cones, or retinal ganglion. Rather than ...that produce sparse labeling would be useful for neuronal tracing experiments where lower levels of labeling...labeling makes it easier to trace a neuron’s projections. Generating animal models: Transgenic animals allow...for retinal gene therapy Find more about precise neuronal targeting using AAV technologies Browse all of... H.P., Krol, J., & Roska, B. (2019). Targeting neuronal and glial cell types with synthetic promoter AAVs...

...Nature’s toolbox. Natural plasmids, also called environmental plasmids, are all around us! A plasmid is a ...through transformation (cellular uptake from the environment), transduction (viral infection by bacteriophages...their host organisms to thrive in competitive environments. The genes encoding these functions form the...less aggressive ways to survive in competitive environments — like by living where no one else wants to....others can’t. This ability to endure a harsh environment is distinct from the benefit of resistance plasmids...lightning-mediated gene transfer in soil. Applied and Environmental Microbiology, 67(8), 3440–3444. https://doi.... environmental plasmids...

... photons are absorbed by matter, they promote electrons within atoms to a higher energy state. This process... called excitation. After a short period, the electrons revert to a lower energy state, whereby a photon...those of absorbed photons. When photons excite electrons of a fluorophore, they can do so at a range of...by the cytometer. Brighter fluorophores have a stronger signal. Table 1: Degrees of brightness for a few...absorbed and then emitted, which creates a relatively stronger — or brighter — signal.   Figure 2: Excitation... real-life advantage of a brighter fluorophore? Strong brightness makes it easier to distinguish a signal...and FITC an organic molecule), their bleed is so strong that these spectra almost overlap. Hence, they ...

...bind to coronavirus spike proteins (Wrapp et al., 2020), which sit on the surface of coronaviruses and enable...injected the spike protein from two different coronaviruses into a llama over several days and isolated ...sybodies that hypothetically contain ones that bind strongly to your target protein. The Seeger lab designed...enrichment step for identifying sybodies that bind strongly and specifically to the protein of interest. The...Structural Basis for Potent Neutralization of Betacoronaviruses by Single-Domain Camelid Antibodies. Cell ... carriers collection Find other COVID-19 and Coronavirus Plasmids & Resources ...

... stem cell-derived neurons expressing ASAP5-Kv. C) Cultured rat hippocampal neuron expressing pan-membrane...synaptic events. Neuron, 112(22), 3680–3696.e8. doi: https://doi.org/10.1016/j.neuron.2024.08.019.  ...ePEmax3 optimized for dicots. NC: nucleocapsid RNA chaperone, RT: reverse transcriptase. Right: T1 tomato plants... gene to confer resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after...antibodies for neurodegeneration research   By Ashley Waldron We recently added two antibodies against important...

...only a few, very strong associations.   Unexpected results Their results showed 96 strongly correlated genetic...accessible to the research community. Sean Misek and Aaron Fultineer in the Beroukhim and Boehm labs pulled...either European or East Asian ancestry, Sean and Aaron knew that their study had significant statistical...mapped to a SNP would be ignored. Working with Lia Petronio and Katherine Huang from Broad’s Pattern team,...varying burden across ancestries. Sean A Misek, Aaron Fultineer, Jeremie Kalfon, Javad Noorbakhsh, Isabella...Isabella Boyle, Joshua Dempster, Lia Petronio, Katherine Huang, Alham Saadat, Thomas Green, Adam Brown,...

...extremely strong affinity for one another. Actually the bond between them is one of the strongest noncovalent...that scientists began to exploit biotin and its strong interaction with avidin for molecular biology and...are useful for biotinylation in different microenvironments. There are also reversible and cleavable biotinylation...attached to a photoreactive arylazide group. Under strong visible light, the arylazide group becomes reactive...Cremer, 2003). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular Biology of Biotin Attachment ... PubMed PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic biotinylation of proteins...

...copy. This is a versatile technique, and is less prone to off-target mutations. Ultimately, successful ...frequently requires considerable optimization to ensure strong counter-selection. Anecdotally, there is also a... evolved specifically for toxicity would allow stronger counter-selection than one merely co-opted toward...rhamnose regulon, we inserted a copy of rhaS with a strong synthetic ribosome binding site (using the online...isothermal assembly master mix. Introduction of the strong transcriptional terminator BBa_B1002 (from IGEM...Modification of Proteobacterial Genomes." Applied and environmental microbiology (2019): AEM-00990. PubMed PMID:...chromosomal gene replacement in Escherichia coli." Appl. Environ. Microbiol. 81.15 (2015): 5103-5114. PubMed PMID...

...neuroscience by granting targeted control over neuronal activity. While they serve similar purposes, both...scale of milliseconds, and can precisely link a neuron's activation to behaviors. Scientists continue to... mutated to have faster off kinetics and allow neurons to repolarize faster.  Chemogenetics does not offer...Administering the drug once can result in several hours of neuronal excitation or inhibition. This sustained activation...tissues, cell types or even subcellular regions of a neuron. Optogenetic approaches excel at discrete cell ...precise spatiotemporal control of a small subset of neurons is necessary, use optogenetics. If broad control...

...monitoring the activity and biochemistry of living neurons in an effort to understand the connection between...available beam splitters. In addition to imaging neurons, Griesbeck adds that this color change has a particular...refined by a secondary screen in rat hippocampal neuron cultures. The optimized Twitch sensors allowed ...researchers to see tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes...transition metal ions, such as zinc, copper, and iron. “These are essential metal ions, but they are also...calcium sensors for functional in vivo imaging of neurons and T lymphocytes. Nat Methods. 2014 Jan 5. doi...

...sterilization as mycoplasma can even escape 0.2-micron filters. Figure 1: Contamination! Tissue culture...shortly. Once they are present in the laboratory environment, they are difficult to eradicate. You can suppress...maintenance of the cell lines. Providing such an environment for the lab facility will not only reduce mycoplasmas... you are careful enough to provide a sterile environment for work, it will soon become a habit before ...separate footwear for the lab to avoid bringing in environmental contaminants.  Figure 2: An Addgenie...lines for assay or analytical purposes. Use 0.1-micron filters as are they are a better way of avoiding...avoiding mycoplasma, rather than using 0.2-micron filters, for media filtration. There is no 100% assurance...

...reversibly degrade proteins of interest fused to a degron, a domain that induces degradation. In plants, ...ligase complex, promoting its interaction with the degron sequence and ultimately its degradation. While ... needed for AID. By tagging proteins with a 7kD degron called mAID and expressing the essential plant ...channels that can be used to silence the activity of neurons. Earlier this year, the Hegemann lab and collaborators... cysteine.  With the introduction of MerMAIDs, neurons can be turned off quickly and with high precision... a valuable optogenetics feature when studying neuronal activity. The researchers identified MerMAIDS ...

...there was strong expression of Cpf1 and GFP in the targeted brain region and ~75% of neurons were co-transduced...Cassettes transfection gRNAs expressed from a polycistronic transcript  Requires co-transfection of Cas9...Cassettes transfection gRNAs expressed from a polycistronic transcript Requires co-transfection of Cas9  ...multiplex edit 3-4 genes in a primary culture of mouse neurons and in vivo. For these experiments, cells were ...-transduced with Cpf1 and GFP.  Of these neurons, ~15% had indels at all 3 targeted loci.  See the chart...

...Murphy’s law definitely applies: what can go wrong, will go wrong. The trash removal service slams dumpsters...useful transferable skill. Being able to stand up in front of an audience and convey your ideas clearly and...project. So whenever you freeze, or when things go wrong, ask yourself, “What next step will help me achieve...term “diving board” so you end up acting it out in front of a very confused group of visiting Chinese undergrads...incorrectly on slides 8-16. Your notes disappear. The wrong version of your slide deck gets uploaded. Your mic...

...Tet-on transgene vector. Degron tags Tagging relevant proteins with a degron tag can provide inducible...removed. Degron tag systems include small molecule assisted shut off (SMASh), auxin-inducible degron (AID)...SMASh degron tagging vector! Destabilizing domains Destabilizing domains (DD) are akin to degron tags,...

... RNA-sequencing to identify neuronal class and deep sequencing of neurons from each class to fully characterize...Addgene.  C. elegans open resources The C. elegans Neuronal Gene Expression Map & Network (CeNGEN) The CeNGEN...comprehensive gene expression atlas of every C.elegans  neuron. Scientists are currently doing this using two ...this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin dependent manner. This...Gene Expression Map of an Entire Nervous System." Neuron 99.3 (2018): 430-433. PubMed PMID: 30092212. PubMed...

...across the membrane of cardiac cells. Similarly, neuronal signaling is carried out by action potentials ...perceived signals to physical actions. Understanding neuronal pathways and the function of certain cells depends...be directed to specific types of cells, such as neuron subtypes, thanks to viral serotype specificity ...available VSDs has been successful even at the single neuron level in vitro. In vivo monitoring has also been...membrane as well. Another flavor of these tools is Voltron, which uses rhodopsin as the voltage sensing component...voltage is particularly physiologically relevant in neurons, myocytes, and endocrine cells. It follows that...

...Chemical Synapses in the Brain. Neuron 68:843–856. https://doi.org/10.1016/j.neuron.2010.11.021  Gambarotto D...techniques like super-resolution microscopy or electron microscopy. Since its introduction in 2015 by ...2010). How do you overcome this resolution limit? Electron Microscopy (EM) is one option, but the equipment...helped researchers gain a deeper understanding of neuronal synapses, centrosomes, parasites, and more (Bertiaux... Ashley Waldron...

...norepinephrine (NE) expressing neurons and GFP (green) to label neurons transduced by each serotype. ...challenge to scientists. What works well in one cell or neuron type may not work well for a neighboring cell type...express ChR2(H134R) in A2 norepinephrine expressing neurons. We tested four serotypes containing an expression...84%) were both excellent at infecting our target neurons (Figure 1). However, this one set of experiments...Perspectives on Data Sharing in Deep Brain Stimulation. Frontiers in Human Neuroscience, 14(December), 1–12. https...