We narrowed to 931 results for: TIM

...Currently, Aggarwal, and colleagues Filip Tomaska and Tim Hanson, are characterizing variants of iGluSnFR3 ...Campus, HHMI where he works on developing and optimizing biosensors in the lab. He has recently developed...sensory input from our extremities. “Here, painful stimuli are processed by a network of excitatory and inhibitory... demonstrate pain response due to blue light stimulation of dual virus-expressing CR neurons. (C) Duration...biosensors.   This screening method allowed Tian to optimize a red‑shifted archaerhodopsin‑derived genetically...

...24336571. PubMed Central PMCID: PMC4089965. 9. Wang, Tim, et al. "Genetic screens in human cells using the...References 1. Cheng, Allen A., Huiming Ding, and Timothy K. Lu. "Enhanced killing of antibiotic-resistant...PMCID: PMC4151723. 2. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize...

...Plants. Carola Engler, Mark Youles, Ramona Gruetzner, Tim-Martin Ehnert, Stefan Werner, Jonathan D. G. Jones...to overcome them, and why she spends some of her time engaging with others on Twitter, among other things... transcripts as the ones that people use all the time. Now there are many, so if you want to make a 10...switching to Golden Gate is a conceptual barrier. Sometimes people don’t know how to start. They might want...Most Sainsbury scientists are active on Twitter. Sometimes it’s a good way to find out what’s happening in...experiences, articles about women in science, and the sometimes stupid or uncomfortable things that have happened...say to our readers? Patron: Remember plants. Every time you look at your plate, think about how much that...

...10.1093/nar/gku936 Eva K. Brinkman, Arne N. Kousholt, Tim Harmsen, Christ Leemans, Tao Chen, Jos Jonkers, Bas...into your target cells with Cas9. Hooray! Now it’s time to make sure your genome edits went according to...within a given genomic window, along with the estimated editing frequency of your sgRNA-Cas9 combo. Pro... from the donor DNA. This method can precisely estimate editing frequency in a bulk population and also...Cas9 or gRNA is the issue, check out or blog on optimizing this step, or consider enriching for transfected...

...Boston Children’s Hospital and Harvard-based lab of Tim Springer, the founder of IPI. Caitlin Faulds is a...

...conjugate or conjugation process. Sodium azide, an antimicrobial agent, can block some amines needed for conjugation...In those cases, thimerosal can be used as an antimicrobial agent. Other chemical interferences with conjugating...measurable light or color in response to a specific stimulus, can be conjugated to antibodies in order to allow...produces a colored product that accumulates over time which is relative to the amount of enzyme present...purchased already conjugated, which can save significant time in the lab. However, if the antibody you want is...ubiquitously expressed protein such as GAPDH or tubulin (sometimes referred to as housekeeping genes). This allows...protein or protein interactions in a specific timeframe or context. Immunoprecipitation (IP) IP uses immobilized...

...neurons upon exposure to light through direct stimulation of ion channels. Naturally occurring channelrhodopsins... nonspecific cation influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely ...been created via genetic point mutations, codon optimization, and chimeric fusion of domains from two different...engineered with enhancements such as: Human codon optimization Example: Halo Increased photocurrent amplitude...activate different neural populations at distinct times. Temporal considerations Temporal precision is key...and promoter system used, there is an incubation time (days to weeks) until peak expression of the opsin...fused to Cry2 and the other half to CIB1, light-stimulated heterodimerization can reconstitute the protein...

... an optimized gRNA for your application (see: gRNA Design Software ). In addition to an optimized gRNA... currently no defined measure of fidelity, so sometimes they will be referred to as increased fidelity...structured motif to enhance pegRNA stability PEmax — optimized PE architecture; increased synergy between PE ...compared to HDR. Prime editing requires more optimization and typically has lower efficiency (< 50%) than...integrase landing site ( attB ) and the other to stimulate DNA repair. This can result in more efficient .... gRNA design for CRISPR libraries is usually optimized to select for gRNAs with high on-target activity...activators and targeting promoter regions. You might sometimes see these referred to together as CRISPRi/a. dCas9...

... that allow you to move around pieces of DNA. Oftentimes several approaches will work for any specific...specific DNA sequence. This can make it difficult at times to identify compatible restriction enzymes that ...behind a short scar in the DNA sequence and can be time-consuming compared to other cloning methods. Read...simplifying the process and reducing the amount of time required for cloning. To utilize this approach, ...enzymes. Thus, you can clone your gene of interest one time into a donor plasmid or acquire one that already...polymerase used, and the single A overhangs degrade over time, further reducing ligation efficiency. TOPO-ready... fragments of DNA in the chosen orientation at a time, and without the need for any unwanted sequence ...

...be low, but generally higher than HDR pegRNA optimization can be challenging Prime Edit Repress or Interfere... and base editor variants often have different optimal editing windows. Be sure to refer to the literature...gRNAs Once your target sequences are selected, it’s time to design your gRNA oligos and clone them into your...and express these components (Figure 2), and the optimal system will likely depend on your cell type. Figure...will work for your experiment) and the expression time (transient or stable). For plasmid and viral-based...best for your experiment. It may be necessary to optimize your delivery conditions, especially if you are... gRNA and Cas enzyme to your target cells, it is time to validate your genome edit. CRISPR editing produces...

...of replication, and an MCS. They are small and optimized to help in the initial cloning of a DNA fragment...field of recombinant DNA technology have been optimized for studying and manipulating genes. For instance...bacterial cell division is reduced, as it takes more time to copy this extra DNA. Because of this reduced ...developed the process termed Sanger sequencing, sometimes referred to as chain-termination sequencing or...Sanger sequencing requires a lot of DNA because the ultimate goal is to have a fluorescently-tagged nucleotide... assemble the whole puzzle. That’s where NGS (sometimes referred to as “second-generation sequencing”)...captures the fluorescent color that is emitted each time a base is added. Again, each base (A,C,T, or G) ...

...viral replication (E4, E2a and VA). These are sometimes combined within the packaging plasmid. For a summary...occur, although at a low frequency which has been estimated to range from 0.01% to 3% depending on cell type... range of cell types in vivo. AAV-DJ has been optimized further for specific cell and tissue types, like... lab has developed plasmids containing SaCas9 optimized for their use in rAAV vectors . Read our blog ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear...protein (DBP) that enhances DNA replication by stimulating elongation and stabilizing the displaced single-stranded...

...27760321. PubMed Central PMCID: PMC5081405. 6. Wang, Tim, et al. "Gene essentiality profiling reveals gene...Karolinska Institutet, in Sweden. It can be very time consuming to design 5 guide RNAs (gRNAs) targeting... failing to upregulate the protein following stimulation would then be separated by sorting, and subsequently...much, showing that we are far from consensus on optimal gRNA design. With Green Listed you can also easily...PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize...

...start sites, such as promoters and enhancers. Many times, integration sites are found in proto-oncogenes.... of helper-free production cell lines also saves time in downstream purification, as extraneous plasmid...different retroviral vectors using a novel, CoDOn-Optimized gene for chimeric GALV envelope. Viruses , 13 ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear...

...size of the DNA removed from the viral genome. Over time, subsequent generations of rAdV vectors have been...needing transgenes to express for a shorter period of time or when needing to package larger cargo. CRISPR-...vectors are also being used in cancer therapy to stimulate anti-tumor immune response in different ways, ... a given virus or infectious agent at a specific time point. Transduction The process of artificially ...

...assess viral integration (see Kim et al. (2021), Timms et al. (2018), and Wells et al. (2020) for examples..., A., & Pour, M. F. (2020). Concise review on optimized methods in production and transduction of lentiviral...https://doi.org/10.1261/rna.2192803 PMID: 12649500 Timms, R. T., Tchasovnikarova, I. A., & Lehner, P. J. ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear...

.... Other molecular targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific...chemical signals such as IL-8, IFNγ, and colony stimulating factor 1 (CSF-1). Additionally, tumor cells recruit...cell-surface proteome to inhibit expression of co-stimulatory molecules needed for T cell effector function... most acclaimed form of immunotherapy in recent times. These engineered T cells are capable of recognizing...immunotherapy targets can be utilized at the same time. A combination of tolerance marker blockade and ...

... polymerase transcribes DNA to mRNA, which is ultimately translated into a functional protein. Thus, the...Tetracycline response element promoter; often optimized for mammalian systems Types of RNA Polymerases...

...conjunction with the MoClo Yeast Toolkit (YTK) to optimize secretion and cell surface display of proteins...are made with zCas9i, a high efficiency intron-optimized SpCas9-coding gene. MoClo CRISPR/Cas Toolkit for...

...antibodies through their HLDA workshops. Let’s use TIM-1 (Single pass type-1 membrane protein), which has... Triton X-100; and methanol, we found that the optimal perm/fix solution depended on the specific antibody...wild-type and the KO population (Figure 1), further optimization can be performed to both maximize the separation...concentration (this can also save money!); and optimizing fixation/permeabilization reagents for intracellular...intracellular targets. Since knockout experiments can be time-consuming and expensive, it may be helpful to find...caused by siRNA is also transient, and as such, the optimal knockdown of the protein target will depend on ... is not efficient at the protein level at the timepoint studied Off-target activities of the RNAi Correctly...

...G-protein and activate neuronal firing through stimulating phospholipase C, which releases intracellular...

...Addgene would also like to thank Viviana Gradinaru and Tim Shay for their support and input. Resources and ... Hudry et al, 2023; Hinderer et al, 2018). The optimal dose for each particular experiment is determined...systemic AAVs are a good fit for your experiment, it’s time to start selecting and testing systemic capsids!...

...receive different types of stimulation may help to refine and optimize your stimulation parameters as well. ...prefer to spend most of its time in the chamber in which it receives the stimulation (conversely, if the circuit... and stimulate them for 30min to an hour and observe what happens. You never know how stimulation of a...personally interested in include: intracranial self-stimulation (ICSS) and place preference. The lab I work in... electrode and non-specifically electrically stimulating an ill-defined region of the brain. With the ...particular hole (the active hole) induces laser stimulation of the chosen brain circuit, while a nose-poke...different patterns and the animal only receives stimulation in one of the two chambers (see image below)....

...quickly Wet transfer Increased time Handcast gel Increased time; have to make as-needed Increase...costs; increased optimization Normalizing to total protein loading Increased time; increased reagents...from lot-to-lot, and have to be re-validated each time a new lot is purchased. Monoclonal antibodies, on...available for your target! Incubation temperature and time Incubating at a lower temperature reduces background...to reduce background while keeping the assay run time to two days.  Indirect or direct detection method...as needed. However, more antibodies mean more optimization and likely higher background noise as well, ... Good for higher weight proteins Long running time   It’s important to note that Bis and Tris gels...

...Reddit AMA Managing time and energy between work and the rest of life is sometimes more of a struggle ... make a change. #2 Thou shalt sometimes put work ahead of life Sometimes, work has to come first. That... for you, but most days go home at the same time. Use time management tools and tactics to plan your work...let my daughter skip school a couple of times. Be sure to add time with your family, for yourself and for...energy and attention. The goal is to maximise the time you are enjoying both work and personal life. There...struggle and sometimes less. When we are lucky, we are “in the zone” and everything seems to fall into place....commandments for making it all work – most of the time. #1 Thou shalt find enjoyment in your work If you...

...reaction optimized. You’ll want to run gradients for optimal annealing temperature and optimal primer concentration...chemistries available and they can be the more optimal choices at times. For example, chemistries that rely on.... The Ultimate qPCR Experiment: Producing Publication Quality, Reproducible Data the First Time, Trends...more in-depth tips on probe chemistries, primer optimization, controls, and analysis.  As a rule of thumb...copy of a transcript. But this sensitivity can sometimes cause as many issues as it solves, so understanding... should be between 100 and 400 nm and must be optimized experimentally.  Probe should have a melting temperature...appropriate reference genes - and it is well worth your time to do one (Biassoni, 2014). At the bench As a highly...

...asked that question today, because I sometimes wish for a stretch of time that doesn’t involve critical thinking...more mentally challenging than lab work at times.” Sometimes a desk job is defined as sitting down at a...how much of that time you’re actually spending doing work. Yes, you may legitimately need to be in the...because of the nature of your time course, but you’ll inevitably have downtime and plenty of it. That’s not...can volunteer their time to share their knowledge with the community (and sometimes conduct their own independent... being the first, and for a very brief period of time, the only, person to know a brand new fact is often...recently who referred to her lab as her family three times during a 10 minute conversation. Leaving a tight-knit...

...block of time and a quiet space where you can address the issue head on. Prepare for this time as if it...need ahead of time and mute your notifications during. (Boyes, 2019) If you’ve taken the time to do this ..., set a timer for five minutes and come up with as many solutions as you can in that timeframe. Tell yourself...you can, set aside time to prepare for future challenges. This doesn’t have to be time-consuming or even...problem-solving process?  You may want to start by carving out time to tackle it. It’s easy to get distracted by an ...hire more people? What would you do with unlimited time? Another strategy for your brainstorm session is...37% higher in originality. It may not be easy to optimize virus production with roller skates—though I could...

...printed your poster and practiced a number of times, it’s time to expertly show off your science.  Prepare...of thought into designing your poster – now it’s time to practice your delivery. Consider your audience...details or omit crucial background. Practice a few times while standing at your poster – it’s a different...unfamiliar venue. Finding your poster board well ahead of time ensures you aren’t scrambling to tack up your poster...before the session begins. Plus, you’ll have extra time to practice at the actual venue. Introduce yourself...to a poster where the presenter spends the entire time talking to the poster, as opposed to engaging with...on the recent literature in your field. Take your time: We often get so nervous when asked a question that...

...Awardees: Xin Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their...our recent blog post. Fatima Enam: Biosensors for human milk oligosaccharides Fatima Enam is currently a...beneficial bacteria in the newborn gut. One of Fatima’s ultimate goals is to engineer bacteria for the efficient...technologies are too time consuming in the first case and nonspecific in the second. Fatima therefore set out...discoveries. Read on to learn more about Xin and Fatima's work and keep checking back in with the Addgene...Light-inducible Proteins, or FLiPs.  Xin later optimized these first generation FLiPs by mutating Dronpa...healthy microbiome. In order to achieve this goal, Fatima first needed an efficient and cost-effective way...