...plasmids: Luc (intronless; CMV-LUC2CP/ARE and Luc-I (intron-containing; CMV-LUC2CP/intron/ARE). The Luc-I...Luc-I intron is a chimeric β-globin/immunoglobulin intron that has been commonly used in constitutive splicing...Multimerization via the SunTag system also shows strong upregulation of gene activation when fused to dCas9... simplifies the process of creating complex polycistronic lentiviruses. The MuLE toolbox contains a series...induced-pluripotent stem cells (iPSCs) to take on a “neuron-like” phenotype when targeting genes involved in...in mammalian cells, yeast, and Drosophila. Synchronization of secretory protein traffic in mammalian cells...disrupts this interaction and thus triggers a synchronous release of the reporters which can then be tracked...

...tobacco. Studying gene function in C. roseus is strongly limited by the availability of methods. Infection...reporter gene examples are below. The GUS gene (β-glucuronidase) with histochemical GUS-staining is a very useful...days in the dark after the transformation. Use introns Plant promoters can be recognized or leaky in Agrobacterium...Agrobacterium, but not the plant. For example, we have seen strong expression of the GUS gene with the commonly used...Agrobacterium. This can be easily avoided by the use of introns, which will be removed from the transcripts in ...plants, check out this interview with the Nicola Patron, who shared the MoClo plant kit via Addgene. For...well working protocol in your lab, it can become a strong tool for your research. Feel free to add further...

...factor RepA binds to the iteron to initiation replication. However, in iteron-containing plasmids,  RepA... regulation system involving antisense RNAs or iterons that inhibits replication when copy number in the...   Figure 1: The replication problem. Iterons are repeated sequences within the origin of replication...del Solar et al., 1998). There are two models for iteron-based plasmid replication control that are not ...the second model, termed the “handcuff” model, iterons of two separate plasmids bind to the same RepA ...these plasmid types? Read our Plasmids 101: Environmental Plasmids blog). Scientists have designed small...replication and transfer systems and host taxonomy. Frontiers in Microbiology 6: . https://doi.org/10.3389/fmicb...

...electrochemical sensors for emerging environmental pollutants.” Front Chem. 2014; 2: 41. PubMed PMID: 25019067...variety of applications, including detection of environmental pollutants, drug discovery, and disease diagnosis...instead of protein-based fluorophores, including: Environments with higher temperatures, since proteins can...Jonathan. “RNA Fluorescence with Light-Up Aptamers.” Front Chem. 2016 Jun 28;4:29. PubMed PMID: 27446908. PubMed...systems using immobilized aptamers.” Biosens Bioelectron. 2011 May 15;26(9):3725-36. PubMed PMID: 21419619.... PubMed Central PMCID: PMC4800036. 12. Wang, Rongsheng E., et al. “Aptamer-Based Fluorescent Biosensors...assay for prion diseases diagnostic”. Procedia Environmental Sciences. Volume 12, Part B, 2012, Pages 1348...

... pSC101 copy number is regulated by iterons. RepA binds to iterons and recruits host proteins to the ori...is initiated at the ori and may or may not be synchronized with the replication of the host cell's chromosomal...RepA binds to directly repeated sequences called iterons located near the ori and directly interacts with...check, high copy numbers can allow RepA to link iterons from two different plasmids, forming a pair of ...either of them (Park et al., 2001). The number of iterons encoded on a given plasmid influences how high ...host, as no additional components are necessary. Iterons are not the only tool to stringently regulate plasmid... numbers, RepA proteins form complexes between iterons on different plasmids in a process called "handcuffing...

...neural fibers developed by the Anikeeva lab are microelectronic fibers made from thermoplastic polymers that...desired thickness. The fibers are drawn with microelectronics included in them for downstream control – ...trended with expression of the opsin in the target neurons, validating the functionality of these inputs and...Success! They also observed reliable optically evoked neuron activity for at least several months after implantation..., P., & Chandrakasan, A. P. (2020). Modular Optoelectronic System for Wireless, Programmable Neuromodulation... Anikeeva, P. (2019). Flexible fiber-based optoelectronics for neural interfaces. Chemical Society Reviews...A., … Anikeeva, P. (2023). Multifunctional microelectronic fibers enable wireless modulation of gut and...

...the first light-activated tool for controlling neuronal activity, channelrhodopsin-2 (ChR2), a blue light-activated... exposed to blue light, this channel activates neuronal activity by opening and allowing positively charged...channels (rhodopsins) either excite or inhibit neuronal activity upon exposure to light within a specific...hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine excitation and inhibition by ... when you want to activate or inhibit specific neurons without needing to consider secondary signaling... off-target activity. Whether you are studying neuronal signaling or cancer pathways there are a number...2022). Red Light Optogenetics in Neuroscience. Frontiers in Cellular Neuroscience, 15, 778900. https://...

... How do you visualize protein turnover within a neuron? For my PhD I studied a synaptic protein that is... to study how protein turnover is regulated in neurons. Protein turnover is the combined effect of protein... I wanted to measure it’s levels in its normal neuronal context. In other words, I needed a method for...for visualizing protein turnover in situ in neurons. The classic approach for studying protein turnover... whole population of one particular protein in neurons or in any other cell type. However, we cannot distinguish... newly synthesized proteins in rat hippocampal neurons." Nature neuroscience 13.7 (2010): 897-905. PubMed...Robert H. Singer. "Single β-actin mRNA detection in neurons reveals a mechanism for regulating its translatability...

...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,...

...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...

...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...

...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...