...us take comfort in the fact that it’s often not quantity, but quality that really matters. Well, it turns...another example, neuronal gene expression by an anterograde AAV was shown to drop off drastically with lower...concentrating your AAV above your desired titer and want to dilute it, err on the side of caution when adding...References 1.Clark, K. Reed. "Recent advances in recombinant adeno-associated virus vector production." Kidney...PMID: 11841606. 2. Schultz BR, Chamberlain JS. Recombinant Adeno-associated Virus Transduction and Integration...PMC2574934. 3. Zingg, Brian, et al. "AAV-mediated anterograde transsynaptic tagging: mapping corticocollicular...

...time, he is pursuing a plant engineering project at BosLab. Once he optimizes plant cell culture and transgenesis...transgenesis he wants to share his knowledge with the community by teaching a DIY plant engineering workshop...and to see old pictures of the lab!). By 2016, we wanted to be taken seriously so we made it official by...ecology but she became a member at BosLab because she wants to engineer microbes. We’ve ordered a few yeast ...

...physiologically-relevant systems, to catch all the important interactions. It’s also important to determine...speak, with collaborators in Paris and New York. We want YOU to join the fight against coronavirus We are... a lot of background in our study - so you might want to be careful with these clones (and help us figure...been quite the administrative headache receiving constant email requests and tracking their fulfillment ...

...people use is the built-in whiteboard, in which participants can contribute and sketch on the board without...control panel on the top of the screen and every participant get to annotate things as they come up in their...time breaks up the monologue a little and gives participants something else to focus on. Use collaborative... creativity are pivotal in writing publications/grants too. As described by several works (Wuchty et al...fades away the longer we are on it – so it is important to take breaks. After 1 hour (or another designated...be done randomly or manually assigning which participants to group together. Breakout Rooms can be used...

...-AAV-syn-jGCaMP7c variant 1513-WPRE (105321-AAV1) pGP-AAV-syn-FLEX-jGCaMP7c variant 1513-WPRE (105322-...vectors to more serotypes. We currently package four variants of these biosensors, which are optimized for specific...relative to baseline fluorescence, as the GCaMP7f variant, and is approximately 5-fold brighter than GCaMP6s...GCaMP6s. jGCaMP7f is the fastest responding variant. jGCaMP7b exhibits the brightest resting fluorescence...fluorescence from inactive neurons is reduced. This variant has an approximately 12-fold increase in brightness...

... mAID and expressing the essential plant protein TIR1 in non-plant cells, which interacts with the cells...Cecchetelli Auxin Inducible Degradation (AID) uses plant hormones called auxins to conditionally and reversibly...a degron, a domain that induces degradation. In plants, auxin binds to TIR1 of the SCF-TIR1 ubiquitin ...engineering SpCas9 Article contributed by Shreya Vedantam CRISPR/Cas9 is widely used genome editing tool...versatility of the engineered SpCas9-NG and its variants can be used to engineer specific sites that were...

...other professors at the University of Barcelona wanted to develop strategies to make our classes more ...thinking about how to solve this problem. I didn’t want to face another group of sleepy students at 8 a....engaging card game could help me get the engagement I wanted. I therefore started planning a game that would...following lessons. And let me tell you more, they had brilliant ideas on how to solve many therapeutic and pre-clinical...educational nature of this material are welcome and if you want to receive our modifiable powerpoint file, please...University of Barcelona. He is also founder and consultant at FRESCI particularly interested in advanced...

...Imaging by Coat protein Knock-off) instead takes advantage of the ribosome’s movement. When the ribosome ... reporter. Delayed translation is known to be important in development, and Halstead et al. used TRICK...mRNA and keeps it in a single field of view for constant tracking. You can then directly monitor multiple...that can recruit GFP fused to the SunTag-specific antibody scFv. Each time the reporter is translated, the...second. They also explored the effects of 5’ UTR variants in translation of cell cycle protein Emi1, showing...and 3’ UTR. Before beginning experiments, it’s important to verify that these changes do not impact translation...

...expression and largely defines the amount of recombinant protein that will be produced. Many common promoters...and steroid regulated promoters commonly used in plant research. Promoter Subtype Promoter Example Activator...promoter is right for me? Here’s a list of some important variables to consider when choosing an inducible...various eukaryotic cell types (mammalian, yeast, plants, etc.) require unique promoters and there is very...’re expressing a gene that may be toxic, it’s important that your inducible promoter not be too leaky....low leakiness. Inducibility level How much do you want to induce transcription of your target gene? The...

...scientists to both quantify Cas9 activity and sgRNA efficiency as well as enrich for mutants through the presence... created CRISPR fluorescent reporter assays to quantify targeted Cas9 cleavage, and repair through homology... in 2017 to enrich and identify desired CRISPR mutants (Wen et al., 2018). SRIRACCHA uses the Piggybac...presence of a fluorescent reporter. Lastly, once a mutant is identified, the PB transposase can easily and...effective gRNA and enrich cells for your CRISPR mutant of interest, give these methods a try.  References...

...diverse libraries of genetic variants. Several methods for generating mutant libraries exist, but none provide...provides a new approach for generating novel genetic variants in bacteria. Read on to learn about the key components...Assisted Continuous Evolution (PACE) method requires constant turbidity of the culture, so bacteria must be ...novel genotypes that result in a spectinomycin resistant phenotype in bacteria. While there are many known... feature which they used to generate bacteria resistant to both spectinomycin and streptomycin, but could...

...Plasmids 101 posts here. Now that we have covered antibiotic selection here at Plasmids 101, we can talk about...amino acids (called the α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation...they should be. Don't rush the process: It is important to give your plates enough time for any intact...identify colonies that not only have your plasmid (antibiotic resistance), but also confirm those plasmids ...α-peptide DNA disruption in the blue-white screen. Antibiotic selection is also used in conjunction with this...select for plasmid-containing cells without using antibiotic resistance. These rely on cell lines susceptible...

...considerations. First, it is important to establish the type of audience you want to reach with your paper... your paper to your preferred  journal, it is important to check the journal’s guidelines to authors and...catalogue numbers and manufacturers’ details for antibodies, plasmids (like your Addgene plasmid #), and ...an identifier (a bit like a GenBank ID) to all antibodies, animal models, and software tools in the Materials...journals and their scientific communities aim to significantly improve the reproducibility and reliability ...

... ask” once you have found someone you admire and want to learn from. Check out Joanne's Reddit AMA Who... colleagues that have the training or skills you want to learn. Mentors can be senior to you or peers ...mentors from your current organization but also important to build relationships outside your organization...confidential topics Broaden your network Take advantage of formal programs A formal mentoring program ...mentoring circle. This type of program has many advantages and I am seeing a lot of success at universities...feedback. The best thing to do to make your mentor want to keep working with you is report back with progress...

...transcription factors with important roles in Multiple Myeloma), it is important to ensure lack of degradation...Remarkably, small molecule degraders provide distinct advantages over pharmacological inhibitors including rapidly... are happy to share aliquots of dTAG-13 in the meantime, please address your requests for dTAG molecules... chimeras For each protein-of-interest, it is important to determine which terminus can accommodate the...context. Immunoblotting is recommended, while quantitative proteomics experiments will provide the best...

.... 1). When you have to tell an engineer that you want the LIMS software to create a barcode for your plate...each given container, when applicable Any other relevant information Prescribing an official name to all... all of our plates, tubes, and racks meant we had fewer errors in our LIMS tracking system. Though you...terminology of a new field. Annotate: When creating important documentation, including SOPs or project-related...Product Manager role at Addgene is “Words are important!”  If you try any of these approaches in your ...propensity for cross team communication, that’s an important soft skill to carry into the next phase of your...

... nuclease-resistant, resulting in less off-target events. While both have similar advantages, bridged ...Nature has evolved ways to prevent degradation of important cellular RNA molecules with post translational...-Cas9, for use in applications like siRNA and antisense oligonucleotides. Some of these synthetic modifications...Frontiers in Genome Editing). This is particularly important in experiments in which viability is being compromised...technology is that most expression systems are constantly “on”. There is no mechanism to turn cutting off...

...on the conversion of non-malignant forms of cancer to invasive and malignant forms and noticed that myoepithelial...stromal cell modifications, stromal cells are also constantly communicating with a tumor, proving once again...malignancy but also incite different organs to become malignant. Exosomes also contain miRNA (micro RNA) and other... case, exosomes may transport miRNA from drug-resistant cells to sensitive ones. Though this process is..., the microenvironment backstage play just as important of a role if not more.  Many thanks to our guest...

...the brain. “We hope that iGluSnFR3 will enable significant steps forward in our understanding of synaptic...Filip Tomaska and Tim Hanson, are characterizing variants of iGluSnFR3 in vivo and have used it to detect...hydropersulfides modulate protein activity and have important roles during oxidative stress. To monitor this...properties. Her method takes a pooled library of mutant sensors expressed in a mammalian cell line engineered...engineered to be electrically excitable. The mutants are co-expressed with a photoconvertible fluorescent...photoconverted, sorted, and sequenced to identify the mutants for further enrichment. Tian notes that this platform...

...high-intensity light for activation which requires implantation of fiber-optic cables into the brain. ...information about parental channelrhodopsins to create recombinant channelrhodopsin sequences. The model first removed...performed well in a tissue culture dish, that doesn’t guarantee their success in vivo. Therefore, the Arnold and...fiber-optic cannulas were still stereotaxically implanted into the mice’s brains so that light could reach...into a hole in their cage. Since dopamine is an important neurotransmitter that promotes reward-motivated... brain may still require fiber-optic cannula implantation, but altering ChRgers so they can be activated...