...molecule into the cell. The fluorescence that is generated allows for qualitative and quantitative measurement...contact with its RNA aptamer. This increased interaction between Corn and its small molecule makes Corn...developed to quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts... transcripts uniquely transcribed by RNA Polymerase III (5S, tRNA and U6) and measuring Corn fluorescence...effect of mTOR inhibition on the activity of RNA Polymerase III. Red and orange fluorescent aptamers Corn...changes in intracellular viral rna." Journal of General Virology 98.5 (2017): 992-1003. PubMed PMID: 28555544...PMC5602116. Song, Wenjiao, et al. "Imaging RNA polymerase III transcription using a photostable RNA–fluorophore...

...of organelles and vesicles, and more. Although several strategies are available to disrupt microtubules...extended to other GTPases. The Chow Lab has also generated a variety of membrane-recruitment optogenetic ...recently developed a full toolbox and workflow for generating new Rho GTPase optogenetic systems (Berlew et...express the other separately, then use blue light to generate CRY2-CIB1 oligomers that trap the fusion protein..., and 100 times the activity of the original, generated using directed evolution (Tei et al., 2023).  ...license.   Just make sure you don’t overdo it! The moderately-enhanced 10x and 30x SuperPLDs avoid dark-state...Krishnan, N., Ononiwu, F., Pal, D., Manikas, J., Taveras, C., Hall, N. A., Silva, J. D., Freshour, J., &...

...these techniques are cumbersome, often requiring several rounds of selection to isolate plants with the ...organisms like Arabidopsis and Medicago truncatula and several crop species including potato, corn, tomato, wheat... plant cells.   This blog post will present a general overview of plant genome engineering using CRISPR... repress target genes in plants using the same general experimental design principles developed in other...systems, multiple gRNAs can be expressed to modify several genomic loci at once (get more information on multiplexing...sequence to enhance targeting to the nucleus, and several codon optimized Cas9 variants have been created... the plasmids described above, Addgene carries several useful CRISPR toolkits for creating plant expression...

...to three) working in your chosen field. This cooperative education experience (co-op) is what has brought...find out that the two hours were split up between several Addgene employees, who all made the daunting task...old) employees with information on how Addgene operates as a team and shows how they fit into the system... on projects there. At this point, I have seen several lab processes and am currently working on projects...are scheduled to take place and employees are generally expected to be available. In addition to these...student at Northwestern University. She is one of several students completing their co-op at Addgene. This...

...highly multiplexed tissue imaging method that uses iterative staining and chemical bleaching to allow for high-resolution...high-quality antibodies and reagents that you know work…preferably before you start your experiment. (And you’re...the IBEX Knowledge-Base, seeking to create an iterative process that is constantly becoming better, with...Ichise, H., Gola, A., Kabat, J., Lowekamp, B., Speranza, E., Croteau, J., Thakur, N., Jonigk, D., Davis...Hernandez, J. M., & Germain, R. N. (2022). IBEX: An iterative immunolabeling and chemical bleaching method for...00644-9 Radtke, A. J., Kandov, E., Lowekamp, B., Speranza, E., Chu, C. J., Gola, A., Thakur, N., Shih, R...

... to the molecule it detects. This change then generates a reporter signal. Reporter signals may be electrochemical...fluoresce; when present, calcium induces CaM/RS20 interaction, thereby changing the FP’s structural organization...M13. When calcium binds, it triggers CaM/M13 interaction that increases FRET transfer between the fluorescent..., and voltage across membranes. These sensors generally follow the same pattern described above - a sensing...withstand a particular solute concentration or temperature? For further advice, see our posts on how to ... Plate-based assays (plate reader) Detect the average amount of fluorescence across a population of cells...

...the relevant literature to understand where the field is today? Have you talked to veterans in the lab ...-to-date with the literature of a couple of well-known journals that discuss general science in the area... timeline? A typical research project can take several years to complete and has multiple benchmarks. ...before even touching a pipetteman. Take the time to generate new ideas and think about them critically before...

...overlapping or nested on a register, then the operation of one recombinase on the register could affect...affect the way downstream recombinases interact with the register- either by re-orienting their recognition...amplified by a particular primer pair. Then, by leveraging qPCR data from multiple primer pairs, one can...designed to have 20-30 nt length, ~60 oC annealing temperature, and ~50% GC content) so that individual parts...not – the state machine design space is highly degenerate and often times there are multiple register designs...serine recombinases have been described in the literature (5, 6) and may be substituted for one another...predictability of RSMs and recombinase-based circuits in general would benefit from a more comprehensive characterization...

...previously yielded valuable enzymes. Famously, Taq polymerase – from Thermus aquaticus bacterium – was discovered...processing environmental samples Since the high temperatures and unstable ground around these hot springs...collection site, we took GPS coordinates, water temperature readings using a heat gun, and pH measurements... continued profiling samples, and years later, several whole genome sequencing projects were launched ...Parvibaculum lavamentivorans, whose optimal growth temperature is 30°C. Find IgnaviCas9 here! What can you do...applications where reagents must be stored at elevated temperatures for long time periods and workflows where CRISPR...

...looking to determine how Protein A and Protein B interact. You read extensive research on the two proteins... both targets in your specimen. You scour the literature and find an ideal antibody against Protein A ...reduce immunogenicity and thereby enhance the therapeutic potential of antibody medicines. In this process...with human antibody sequences.  Additional considerations Although isotype conversion is routinely used...utility of recombinant antibody tools, the resulting chimera may have different binding properties (Morelock...needs to be addressed when multiplexing. It is imperative that you use narrowly selective isotype-specific...

...Thailand, our annual operating budget is approximately 20,000 USD and we are working on several genetic diseases...: This year, the BeHEARD (Helping Empower and Accelerate Research Discoveries) Award attracted submissions...affected by AHC suffer from recurrent attacks of unilateral paralysis (hemiplegia) that are often painful...stressors, including emotional stress or changes in temperature.  “The disease was first described in 1971,” ...elucidating association between SOX10 and WS4 and also several other rare genetic diseases. We hope kids in Thailand...

...outcrossed knock-in ready for experimentation. In 2014, several manuscripts published in Genetics detailed approaches... DNA as repair templates which reduces time to generate edits to only four days (Paix et al., 2017). The... demonstrated that selection for repair of a temperature-sensitive pha-1 mutation significantly enriches...design The Meyer lab meticulously characterized several aspects of editing. First, they devised guidelines...modifying the 5’ modification of oligos used to generate linear dsDNA repair templates by PCR boosted efficiency...from a single injected animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding...elegans The Meyer and Mello labs also identified several experimental set ups that could improve CRISPR ...

...development of useful diagnostic methods and improved therapies for PZA-resistant Mycobacteria tuberculosis. Investigation...to tag individual strains of bacteria. Their interactions and growth within the biofilms will then be ... taraxacifolia leaves are widely used to treat several diseases. Her goal is to identify and isolate pure...Therefore, understanding neuronal networks, their interactions and information processing during this condition... to get new insights into functional network interactions. Combining synthetic biology with engineering...resources and microfabrication technologies to generate a self renewable sensor, a bacterium that can ...

...for a higher number of MPE. Therapeutic potential  To explore the therapeutic potential of the DAP array...pathogen. Over the past decade, CRISPR tools have leveraged such host-defense mechanisms to enable multiplex...to develop a strategy that would independently generate multiple gRNAs for nCas9-BE, or any other kinds...drive-and-process.” To optimize DAP use, they developed a generalizable method to acquire the most effective DAP array... demonstrated up to 31-loci MBE with over 50% averaging editing efficiency in human cells. MBE with DAP...

...remote collaborations Transferable Skills Guide: External collaborations Transferable Skills Guide: Teamwork...how to seek out such an arrangement, and how to operate once in that position. What is a visiting scientist...new placement. Make sure you read any relevant literature for the techniques you will be performing to ...home institute. Discuss rights Do you expect to generate data that will contribute to a publication? A ...authorship and rights for anything that could be generated during the visit. If you don’t feel comfortable...

...part of my job, in addition to preparing DNA and generating/replenishing kits, is to find new ways to make...extract libraries from previously fragmented DNA.The general process for library preparation using this kit ...total of about 3 hours. It was robust, capable of generating a wide range of insert sizes and also produced...participants from different biotech companies to interact and learn more about one another. They also provided...representatives. I got to talk with a technician from Neon Therapeutics, a biotechnology company that uses neoantigen...extremely helpful since she was familiar with library generation. Also, I now know what to expect at a workshop...

...2012). The polarizing impact of science literacy and numeracy on perceived climate change risks. Nature... between what I was doing in the lab and the interactions I was having outside of the “ivory tower.” I... acute “adeolitis” – received the best ratings overall, simply because none of the participants had preconceived...show that people with greater levels of science literacy are especially prone to this, possibly because...Fischhoff, B. (2017). Individuals with greater science literacy and education have more polarized beliefs on controversial...

...proteomics--the study of the distribution and interactions of proteins within these structures. Enzyme-...substrate to map the spatiotemporal distribution and interaction networks of specific proteins in living cells...process is much slower and can take as long as several days. In 2015, the laboratory of long-time Addgene..., the enzymes function across a much broader temperature range than BioID, which only functions optimally...modular domains that facilitate protein-protein interactions and mediate biochemical processes. One of the...than previously thought. Sidhu’s group has made several hundred SH3 domain containing plasmids available...

... In each case, we leveraged the team’s knowledge of the scientific domain in general and of our data in... that we needed to improve more than that: the general “findability” of the information on the site. Different...approached the issue from multiple angles: the overall user experience of our website, the different website...resources and handle millions of queries. In an iterative process driven by our user experience experts,...approach much more than if we were dealing with general purpose information. Below are a few of the strategies...

... these interactions, and can give valuable insight into DNA expression and protein interactions, including...technique that provides insight into protein:DNA interactions. ChIP works by using antibodies to capture protein... focuses on the DNA your protein is actively interacting with, reducing background information. Your options...could mean needing to increase crosslinking to counteract the effects of increased sample agitation; a ... this investment can well be worth it once you generate your data. However, it may be a good idea to add...