... important proof of concept for future preclinical and clinical work. Find Plasmids for Disease Research...research, researchers are eager to apply it in a clinical setting. CRISPR is often used for animal germline...exon skipping is a good option for DMD treatment. Clinical trials have used oligonucleotide exon skipping...rate of exon 51, which resulted in appropriately localized dystrophin. In a subsequent study, they used CRISPR...six months. Future directions and obstacles to clinical CRISPR editing Given the success of the CRISPR-mediated...be evaluated separately, increasing the time to clinical approval. Even with the challenges precision editing...become one of the first diseases to be treated clinically with CRISPR. Many obstacles remain for CRISPR...

...generally linked to dopamine (DA), a well-known reward chemical and neurotransmitter. But aside from DA’s famous...specific dopaminergic neurons in a brain region called the substantia nigra, which is the main source ... it can be performed in a live awake animal by surgical implantation of the probe into a specific brain... animals (Patriarchi et al., 2018). Similar to calcium and glutamate sensors, the new dLight1 sensor is...in freely moving mice Record DA dynamics of optogenetically activated and inhibited neurons in live mice...neuronal imaging of dopamine dynamics with designed genetically encoded sensors." Science 360.6396 (2018): eaat4422... PMCID: PMC6287765. Sun, Fangmiao, et al. "A genetically encoded fluorescent sensor enables rapid and ...

...swimming where they periodically beat their tails left and right in a symmetrical motion to move forward...behavioral studies. In neuroscience, opsins are typically ion channels that allow charged particles to cross.... The non-specific cation channel ChR2 and the calcium channel LiGluR both cause ion movement that results...Kolmer-Agduhr cells was sufficient to trigger the symmetrical tail movement unique to spontaneous slow forward... photostimulation in order to drive the full symmetrical response.  The function of Kolmer-Agduhr cells...Aristides B., Filippo Del Bene, and Herwig Baier. "Optical control of zebrafish behavior with halorhodopsin...

...to mind when we think “biomedical research". The family entered the biomedical field’s spotlight after...ways these animals have impacted humans through biomedical research. The Camelid family is composed of ...chromobodies,” which are single-domain antibodies genetically fused to fluorescent proteins and used to visualize...therapeutic was approved in 2018, with many others in clinical trials (Jin, et al. 2023). In addition to therapeutics...) (Xu, et al., 2021).      Figure 2: Genetically modified mice offer an alternative method to ...Arbabi-Ghahroudi M (2017) Camelid Single-Domain Antibodies: Historical Perspective and Future Outlook. Front Immunol...

...strengthened my critical thinking and problem solving abilities, increased my technical know-how, and helped...scientist.” For jobs away from the bench, look for “technical or product support specialist,” “field application...application scientist,” or “scientific or technical writer,” among others. Don’t discount job postings looking... you’re interested in (i.e. biotechnology, pharmaceuticals, etc.) Do their jobs sound interesting? Is ...deeper foundation of scientific knowledge and technical know-how that increased my autonomy and satisfaction...from the lab to a desk, although there’s often technical career tracks for PhDs who want to stay at the...

...Bryan Roth’s lab created a collection of plasmids called TRUPATH to interrogate 14 of the 16 non-visual ...TRUPATH collection allows researchers to measure the physical association of combinations of different Gα, Gβ...acceptor. TRUPATH uses an updated version of BRET called BRET2. Figure 1: TRUPATH measures heterotrimeric...N-terminus of the protein. For the Gα constructs, localization of the Rluc8 was more challenging. The Roth ...experimental refinement to determine the optimal localization of the Rluc8 insertion within the Gα. Despite...transducers and that the potency and efficacy is typically non-uniform across the transducerome for a specific...well known and, as a result, a target of many pharmaceutical drugs, there is still only a partial understanding...

...pAAV-hSyn-DIO-EGFP   Biosensor AAV Calcium sensors Tracking calcium dynamics that govern such activities... from Christopher Harvey’s lab. These plasmids localize opsins to the soma to avoid off target photostimulation...need to determine these transduction patterns empirically for their own experiments. To this end, we offer...studying neural networks. We have expanded our calcium sensor collection to include our first Flp-dependent...Several new options are also available for the calcium sensor jGCaMP7 from Janelia, including three jGCaMP7...technology, see our blog post Overview of AAV Encoded Calcium Sensors). Plasmid Serotype Name 104489 AAVrg...

...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...

... alumni as you can or schedule video/calls if they are not local. Alumni meetings can be the most informative...alumni, don’t ever disregard warning signs of unethical behavior, harassment or bullying. Even with the...innuendo and sarcasm, persistent teasing, name calling, insults, intimidation, sexual harassment Isolation... including preventing access to opportunities, physical or social isolation, withholding necessary information...administrative responsibilities? Is the work (topic and/or technical aspects) applicable to industry if this path interests...Questions to ask: Will someone(s) train you technically? Will the PI/supervisor look at your data and... if you take the time to evaluate the options critically. How will it feel when you find it? If you find...

...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...

...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.,...

...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 ...

...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...

...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...