...bacteria) may be the most numerous and most diverse biological entities on our planet, but we still know collectively... refractory to enzymatic digestion, however, a physical fragmentation strategy is preferable. Nebulization...tank with a pressure regulator. Other methods for physical shearing exist as well, such as sonication and...transform your resultant plasmid libraries into chemically competent E. coli and analyze transformants for...reduce the number of cells that will bind non-specifically to either the membrane itself or to any “sticky... antibody or other protein on hand that will specifically bind your target organism, it can be helpful...2015): 185-191. PubMed PMID: 25708933. 2. Cooper, Callum J., Mohammadali Khan Mirzaei, and Anders S. Nilsson...

...to actually hold all those seemingly abstract biological reactions that I learned about from textbooks...physics class to learn about electronics or mix chemicals together in chemistry to see what happens, but...proteins outside of a cell - cell free extracts Typically, if you want to make a protein – say green fluorescent...incorporated into a host cell (through a process called transformation), essentially giving the cell all...they can see, enzymes that can convert certain chemicals into fragrances that they can smell, or enzymes...central dogma while also giving them practice with practical lab skills like developing a hypothesis, precisions...Ally Huang is a 4th year PhD student at MIT in Biological Engineering and is interested in developing synthetic...

...taraxacifolia (Willd), an annual herb of Western Tropical Africa typically known as wild lettuce. Aside from their...researchers in resource-limited settings. More specifically, the Addgene-Seeding Labs plasmid grant provide...ameliorate diabetics nephropathy, one of the major pathophysiological complications of diabetes mellitus. Recombinant...flow. Novel treatment options, as well as non-pharmacological solutions to modify the course of this condition...scanning microscope will allow Tsakanova to image cortical neurons and study changes in neural activity after...working in a wide range of research fields and geographical locations. In addition to providing reagents...

...agreement on the transfer of biological materials in 1995: the UBMTA. Because biological materials are widely ...many different flavors of MTAs, but the Uniform Biological Material Transfer Agreement (UBMTA) is one of...process was time-consuming and lengthy. So the NIH called a meeting in 1990 to discuss the process, which...MTA for that plasmid facilitated by Addgene — basically, instead of having to sign individual MTAs for...agreements allow for centralized tracking when biological reagents are shared — both at what time and with... options for the terms you want to share your biological materials under! While most plasmids at Addgene...

...neuronal membrane trafficking and localization. Improved photophysical properties The improved variants...iGluSnFR3 variants that are more sensitive and better localized for synaptic glutamate response than its predecessors...researchers did the following: Improve sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic...Supplementary Figure 2.   Improved trafficking and localization through different C-terminal anchoring domains...Blog Learn about other Fluorescent Biosensors A Practical Approach to Choosing the B(right)est Fluorescent...indicators with improved activation kinetics and localiation for imaging synaptic transmission” bioRxiv (2022...

... like writing. It’s basically the equivalent of crossing off days on the calendar after completing your... Writing schedule tools How to write a lot: A practical guide by Paula J. Silvia  This book has tips for...take a 5-minute break. Each 30-minute interval is called a “Pomodoro.” After four Pomodoros, you can take...suggestions Hemmngway makes (it wasn’t designed for technical writing), it’s great at flagging passive voice...jargon, this tool can help you avoid overusing technical terms when writing for a general audience. The...goal of the phrase, such as being cautious, being critical, or defining terms. The phrases in Phrasebank ...

...physiology. Ribosome profiling Ribosome profiling, a biochemical method developed in 2009, takes a “snapshot” ...Recent work has adapted this technique to monitor local translation and determine if proteins are targeted...translation While ribosome profiling co-opts the physical location of the ribosome, TRICK (Translating RNA... the reporter. Because it contains a nuclear localization signal, NLS-PCP-GFP then returns to the nucleus...recruit scFV-GFP, generating green puncta that colocalize with the red reporter mRNA. Yan et al. used harringtonine...inhibitor that stalls ribosomes after initiation, to calculate a ribosome translocation rate of ~3.5 codons/second...studying translation in various cell types and physiological states. References 1. Ingolia, Nicholas T.,...

...presented chronologically. With a resume however, most often people choose a reversed chronological style ... job asks for experience in “statistical analysis”, mention “statistical analysis” (word by word) in your... applying for. Also remember that a resume is typically no more than 2 pages long. Write your resume for...handled large data sets and are familiar with statistical analysis. If you have done fluorescent imaging...Some universities even have officers working specifically to assist graduate students in job searching...

...Luciferases are a class of enzymes capable of catalyzing chemical reactions in living organisms resulting in the... is measured resulting from the catalysis of a chemical reaction requiring luciferin, ATP and oxygen as...Fluorescent Protein (GFP) because of the nature of the chemical reaction compared to using a high-intensity laser...Luminescence Fluorescence Source of Emitted Photons Chemical reaction High-energy photons Kinetics of Photon...High-throughput assays Improving Well-established Biological sources of luciferase Although luciferase isolated...with a peak emission at 480nm upon catalyzing a chemical reaction with the substrate coelenterazine and...expressing luciferase Luciferase reporter assay The typical luciferase-containing plasmid is most often used...

...all. Base editing to the rescue! Base editors chemically modify DNA bases without breaking the DNA backbone...elements. They are, broadly: A Cas protein (catalytically dead or nickase) fused to a deaminase that makes...al., 2016). Target-AID acts similarly but not identically to BE3, modifying a 3-5 base window 18 bases ...expressed well. The Liu lab modified the nuclear localization signals and codon usage BE4 to create BE4max...al., 2023; Neugebauer et al., 2023). These CBEs (called Td-CBEs, CBE-Ts, or TadCBEs) are smaller and less...an adenine base editor with improved nuclear localization and expression. This base editor was named ABEmax...Keith Joung’s lab created a dual-deaminase editor called SPACE (synchronous programmable adenine and cytosine...

...systems are available at Addgene! Chemically inducible promoters Chemically regulated promoters are among ...sensitive expression systems are typically less leaky than chemically induced promoters; they show near-zero...transcription is turned ON. Types of inducible promoters Chemical agents, temperature, and light are all examples...decreasing promoter leakiness.   Other examples of chemically induced promoters include positive inducible ... promoters only work in prokaryotic cells and typically only in the same or closely related species from...

...and are often impractical for small projects. Most researchers instead turn to so-called “mismatch nuclease...involves splitting up a sample into thousands of physically isolated partitions (such as droplets or wells...Since there are many thousands of partitions, statistical modelling can be used to determine the actual..., does not require standard curves, and can theoretically detect a single target molecule in a sample....wild type target in each sample can be quickly calculated (details in [2]) and can be used to quantify ...mutations versus those with the bi-allelic mutations typically desired when screening clones for complete functional...

...Immunity, as well as various combinations thereof (so called “Immuno-Oncology”) is exploding. Researchers are... in the presentation, starvation or the use of caloric restriction mimetics trigger autophagy and, in ...Binghamton University! Cancer therapeutics The so called field of Immuno-Oncology, a sizzling hot topic,... and correlating them with tumor mutations and clinical responses. This platform has the potential to ...the use of optogenetic tools is expanding to immunological research. Here the Zhou group presented Opto-CRAC...cells that are not excitable but responsive to calcium. We’d love to expand our cancer and immunology ... References: 1. Pietrocola, Federico, et al. "Caloric restriction mimetics enhance anticancer immunosurveillance...

...emerging engineering technology. Monoculture and chemical spraying, mainstays of modern agriculture, are... by developing a plant biotechnology product specifically for consumer sales. Flowers that change color...a new conversation about GMOs.  Challenge #2: Technical obstacles to plant synthetic biology In addition...understanding of plant biology is a significant technical obstacle to plant synthetic biology. We cannot...creative problem solving like the Howard Hughes Medical Institute have begun to pivot towards plant biotechnology...organism for plant research for decades, and the biochemical pathways involved in central anthocyanin metabolism... – our project wouldn’t be possible. A second technical hurdle is the lack of resources associated with...

...mammalian cells often relied on chemical (CaCl2, cationic liposome) or electrical (electric current forming ...research since their first use in the early 90’s to genetically modify primary cells. Amongst the different vectors...its safety was a disadvantage particularly for clinical development. In addition, MoMLV is inefficient...lentiviral vectors was developed for safe use in a clinical setting. In this generation, the HIV Tat gene ...lentiviral vectors can be used for research and for clinical purposes, however improving the safety of these...than with the 2nd generation. Knowing this you can scale up your production set-up and purify and concentrate...virus in order to aid viral binding. This method, called spinoculation, has been found very effective for...

...HE, Lee C-YS, Calidas D, Lu T, Smith J, Schmidt H, Krause MW, Seydoux G (2014) Scalable and Versatile ...is an assistant professor at the University of California, Santa Cruz. Emerging CRISPR/Cas9 editing technologies...melting and rapidly cooling dsDNA repair templates dramatically boosted editing efficiency (Ghanta et al., 2020... more efficient. Pure PCR repair templates are critical for efficient knock-ins (Ghanta et al., 2020)....Farboud et al., 2019). Work from the Mello lab has dramatically improved RNP editing with linear repair templates...is an assistant professor at the University of California, Santa Cruz where his group focuses on gene regulation...Tzur YB, Friedland AE, Nadarajan S, Church GM, Calarco JA, Colaiácovo MP (2013) Heritable Custom Genomic...

...simulate physiological gene expression regimes that cannot be easily achieved with classical gene overexpression... super-resolution microscopy.” Endosomes are historically difficult to study in living cells using conventional...system to study epigenetic modulation. While catalytically inactive cas9 (e.g. dCas9) has been fused to...activate gene expression, sometimes incorporating chemically inducible approaches to provide temporal control...kinetics of chromatin regulation and subsequent mathematical modeling. Its reversibility allows precise and...hER or progesterone for hPR - leads to nuclear localization, binding to the appropriate promoter sequence...

...Therefore, it is not possible to visualize the localization of protein synthesis. Alternatively, fluorescent...graduate studies I attended a scientific symposium called Horizons in Molecular Biology. This annual symposium...PhD students at the Max Planck Institute for Biophysical Chemistry, Göttingen in Germany. I attended this... symposium seeking inspiration to resolve the technical hurtle of monitoring protein turnover of specific...interesting talk by Prof. Erin M Schuman, a former Caltech researcher, now Director at the Max Planck Institute...technique applies metabolic labeling using a noncanonical amino acid, azidohomoalanine (AHA). In the technique...application of AHA labeling is known as fluorescence noncanonical amino acid tagging (FUNCAT, Dieterich et al....

... The previous post in this series described a practical approach to selecting a bright fluorescent protein...amounts of excitation power (Shaner et al, 2008). In confocal laser scanning microscopy, fluorescent proteins...selective plane illumination, TIRF or spinning disk confocal use entirely different excitation power regimes...performed with soluble fluorescent proteins or localized fusion proteins and do not require dedicated plasmids...Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes. You can follow him...(University of Amsterdam) where she develops genetically encoded FRET based biosensors.     Additional...Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire Fluorescent Protein Collection...

... the flick of a light switch. Neurons are not typically persuaded to fire when light is shined on them...channelrhodopsins. Their new channelrhodopsins, called ChRgers, make the brain a little easier to access...determine membrane expression and plasma membrane localization of transmembrane proteins, like rhodopsins, ...directed evolution, because channelrhodopsins are typically evaluated with low-throughput methods such as ...properties of a channelrhodopsin, such as cell localization, kinetics, and spectral properties, must be ... midbrain, fiber-optic cannulas were still stereotaxically implanted into the mice’s brains so that light...optogenetics.  TL;DR The new channelrhodopsins, called ChRgers, make it easier and less invasive to use...