We narrowed to 842 results for: Eras

...base-pair mCherry insertion. Editing of these motifs generates lesions that are subject to excision and ssDNA...This edit changes the histidine to a tyrosine generating a GFP variant (GFPY66) that has a shifted emission...Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance reporter. Image...hypothesis with GFP fluorescence. To do this they generated a silent GFP construct, that contains an ACG start... of different reporters including mScarlet-I, luciferase, antibiotic resistance markers, and other enzymes...fluorescent proteins, antibiotic resistance, and luciferase. References Gaudelli NM, Komor AC, Rees HA,...CRISPR/Cas9-mediated Genome Editing. Molecular Therapy - Nucleic Acids 5:e334 . https://doi.org/10.1038...

... the ASM General Meeting (now Microbe) in 2013 in Denver, CO. While I had been to the General Meeting ...the next generation of scientists. The connection between mentors and Singletons can accelerate the networking...meet someone new, as they’d been introduced to several people by their mentor, as well as their bin-mates...recognized by meeting organizers in their town hall.  Iterative improvement - Making Binning Singletons better...Goldschmidt conference, Coastal & Estuarine Research Federation (CERF) Biennial Conference, Gordon Research Conference...

...process to reduce viral order fulfilment from an average of 40 minutes to 10 minutes. While users can’t ...result in less than optimal user experience and the overall soundness of the software design. Each of us brings...called Scrum. The Scrum framework is based on an iterative and collaborative approach to development. Scrum...capacity to finish all of the work. Since we work in iterative chunks, we have the flexibility to adapt and easily...allows for life to happen without impacting the overall work. Each of our sprints is themed and the stories... being maintained (i.e. hasn’t been updated in several years) or they make significant changes that make...that helps our users focus on the fun stuff, accelerating research and discovery. We work with transparency...

...opportunity to discover and evolve your transferable skills. Transferable skills are the non-technical skills... 5 o’clock. You’re going to have to talk and interact with...non-scientists! Maybe you’ve just spent... Wu in graphic design. You could dread these interactions, but you could also embrace them. Talking about...production, I didn’t really start to recognize my transferable (or “soft”) skills until I was out of the lab...lab to think about these--get started with our Transferable Skills Guide.     Over the last decade, my... blog Visit our careers topic page Read our transferable skills guide Read blog posts about career readiness...

... slim chance of generating indels with base editors, keep in mind that the generation of indel sometimes...minimizes the second outcome and increases the generation of the desired T-A base pair. Figure 1...inosine), which is treated as ‘G’ by cellular polymerases. Subsequently, an original genomic A-T base pair...simultaneously.     These binary vectors could be used to generate stable genome edited plants via Agrobacterium-...transformation. Cytosine base editors for plant There are several CBEs available for plant systems. Target-AID based... transient assay. For stable transformation, regenerated plants may be subjected to genotyping. If successful...

...means they can be used with or as fluorophores in several different ways. Researchers have used oligonucleotides...can be created in many different ways, but the general premise is that an aptamer in its unbound state...cells. The new aptamer also has a higher melting temperature than its Spinach ancestors, and has a lower magnesium...fluorophores, including: Environments with higher temperatures, since proteins can denature while aptamers ...likely to trigger immunoresponses. Nucleotides, in general, are far less immunogenic than proteins. Assays...ions or particularly small molecules that do not generate an immune response. Aptamers can be selected for...Peixuan AND Haque, Farzin. RNA Nanotechnology and Therapeutics. Boca Raton: CRC Press, 2014. 5. Hayat, Akhtar...

...company that is developing RNAi therapeutics to treat neurodegenerative diseases. He has earned a B.S. ...s Board of Directors, Bart Newland. Newland is General Counsel at Atalanta, an early-stage biotech company...like? BN: I've made the transition now to being general counsel of a small start-up. For the last 20 years...and Biogen. They’re very different jobs. Being general counsel at a small start-up is very interesting...interesting. It’s very varied. There’s IP work to do, general corporate work, and negotiating contracts with other...

...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., &...

...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...

... genes with potentially harmful mutations in therapeutic settings. Silencing of a target gene is achieved...RNA (shRNA) Short interfering RNA (siRNA) Piwi-interacting RNA (piRNA) miRNA and piRNA can be found endogenously...experiments. shRNA plasmids typically use RNA polymerase III promoters, which transcribe small RNAs. The..., shRNA plasmids are being designed using RNA polymerase II promoters. The design of these plasmids is...plasmids One of the most common uses of shRNAs is to generate stable knockout or knockdown cell lines. In fact...collections/mammalianrnai/","type":"EXTERNAL"},"user_generated_content":false}{% endraw %}{% end_module_attribute...(2004). MicroRNA genes are transcribed by RNA polymerase II. The EMBO Journal, 23(20), 4051–4060. https...

...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...

...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 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...

...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...

... 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...

...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...

...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...

...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 ...

...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...

...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 ...

...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...

...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...

... systems that measure biological phenomena by generating an output which is directly proportionate to ...antibody, enzyme, DNA, etc. The receptors job is to generate a quantitative signal upon recognizing the analyte... (fluorescence, pH, etc.). The transducer will generate a numerical value which represents the amount ...disciplines! Most of the sensors designed for calcium generate a fluorescent output, which will turn on, change... It’s a good thing, though! Viral delivery has several advantages over other mechanisms. For one, many...permanently integrate and/or express long term (up to several years!). Biosensors tend to be more reproducible...

...more effectively used in a technology or as a therapeutic.  A (very) brief history GCE came onto the synthetic...specific to the ncAA and orthogonal to (i.e. non-interacting with) the cells’ own natural translation machinery...system (including media, cell type, and growth temperature) may need to be individually optimized for each...focused on turning GCE tools developed in the literature into robust and universal techniques that researchers...fulfill their mission. While they’ve shortlisted several protocols, including their phosphoserine ncAA protocol...., Griffith, M. C., & Schultz, P. G. (1989). A General Method for Site-specific Incorporation of Unnatural...

... 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...

...synthetic) pFZ19 (yellow flame) Temperature inducible promoters Temperature sensitive expression systems ...Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region ...Types of inducible promoters Chemical agents, temperature, and light are all examples of factors that can...promoters; they show near-zero expression at regular temperatures but can be induced by heat or cold exposure....since expression can be repressed by glucose. Temperature inducible promoters are also known for their ...

...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...