...catch our April AAV webinar with Tim Miles, PhD, Director of the CLOVER Center at CalTech? If so, you may...trans-protein splicing. Note, however, that this will require co-delivery of two AAVs at a high MOI, often resulting...1x1012 vg/mouse is more standard) and reflects a desire for very high MOI on top of very high coverage....

...label your favorite protein! While the target only requires a small epitope tag (FLAG is eight amino acids... is advantageous to Cas9’s larger expression requirements (Figure 1), but Cas12a has generally been underutilized...when encoding multiple guides. While two guides require two arrays for Cas9, two guides can be streamlined...

...Passion? A love of learning? No. Or at least not entirely. What truly unites us all is the frustration of... to the day before, which got rid of the issue entirely. In many cases, the answer isn't to work harder...ability to focus, especially on days when I was tired, stressed, or rushing. I would spend the time quietly...

...tropism. Briefly, different envelope plasmids can direct the production of virus with various tropisms.  ...therefore, reduces the number of plasmids that are required in the transfection step of the viral packaging...incompetent? Yes, as with lentiviruses, the genes required for γ-retroviral packaging and transduction are...

...biological phenomena by generating an output which is directly proportionate to the amount of a given factor ...either factors as proxies for a cellular process or directly measure a cellular process. Cellular phenomena...type of cells the viral particles infect. This can direct your biosensor to specific tissues or cell (sub...

...transfect’, such as primary cells. They can also be directly injected into hard-to-reach organs, such as the...like replication incompetent viral particles, are required. We will be covering these safety measures in ...dividing cells, as their access to the host genome requires breakdown of the nuclear envelope, which occurs...

...truly believes that sharing speeds science, and our entire mission is to enable that process. Our repository... made most of the Antibody Guide figures. This requires a fair amount of communication between our content...of date as techniques and knowledge changes and require updating or retiring. CRISPR/Cas9, for example,...

...because “it kinda sucked to work remotely. I was also tired of the contracting life where you do a little bit... was helping cover social media while the Hutch hired a full-time social media manager. “I usually start...magazine. Getting pulled in all of these different directions is sometimes at odds with the space needed to...

...RNA, a fast, strong protein denaturant would be required- something that broke down RNases before RNases... effective at isolating total RNA, this method requires a lot of time and depending on how many samples...without a kit!) As mentioned above, working with RNA requires keeping your samples cold until homogenization...

...to read a formulaic fiction story that doesn’t require any detailed retention of information or formulation...something verbally. Sometimes a group was just really tired and needed a slower, simpler presentation than I... I had planned; other times students were fired up and enthusiastic and needed a faster presentation with...

...Much of today's biological research requires a close examination of specific proteins within a system...tags, however, tended to be very large and often required the incorporation of a cleavage site and a post-purification...no cut or dried answer. Successful purification requires the tag to be accessible to the metal ions once...

...sufficient for certain patterning activities of entire tissue types or even organs. In frogs, ectopic ...electrical communication is briefly inhibited in the entire worm during regeneration, the worm creates two ... artificial intelligence," says Michael Levin, Director of the Allen Discovery Center. The future of this...

...sex, the National Institutes of Health (NIH) now requires that sex be included as a biological variable ...variable in all NIH-funded research. Janine Clayton, Director of the Office of Research on Women’s Health, emphasizes...gender in science and technology and currently directs the EU/US Gendered Innovations in Science, Health...

...with multiple inserts or internal mutations that require more than two sequencing reactions. In the more...to ensure uniformity of the culture. Sequencing directly from the glycerol stock saves us the step of purifying... Each sequencing service will have different requirements for volume and concentration, and not all providers...

...new job skill? Finish your thesis? Knowing your desired outcomes will help you assess if the coach you ...coach usually will be a clear communicator and make direct suggestions for improvement and change. Work ...relationship has been successful – what are the desired outcomes? Ask openly for clear information on...

...asymmetry in the core sequence gives the loxP site directionality, and the canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT...reversed.   Deletion: If the sites face in the same direction, the sequence between the loxP sites is excised...cerevisiae FLP-FRT recombination system is another site-directed recombination technology very conceptually similar...

...photon 2. Energy transfer from the excited donor directly to the acceptor fluorophore--think of it as a ...fusions varies with transfection efficiency and any unpaired fluorescent proteins can contribute additional...Check Out Fluorescent Protein Biosensors Browse the Entire Fluorescent Protein Collection ...

...effectively utilize the host cell’s inherent homology directed repair (HDR) process following the double strand...break and introducing the tag through homology directed repair To introduce the fluorescent tag to the...on the Addgene Blog Learn More about Homology Directed Repair Learn More about Non-Homologous End Joining...

...laboratory, a high level of experimental control is required, because seemingly tiny external factors can have...overexpress a gene that alters neuronal function directly such as receptor subunits or neurotrophic factors...smaller and may be preferable if wider diffusion is required. On the other hand, lentiviral vectors are superior...

...Development Dynamics Database (WDDD) The WDDD aims to acquire quantitative data on all the cell divisions of ...Kit for CRISPR Cas9 edits At Addgene, we have the entire SapTrap toolkit from the Jorgensen lab. SapTrap...Project: The Complete Gene Expression Map of an Entire Nervous System." Neuron 99.3 (2018): 430-433. PubMed...

..., comprised of PAS and GAF domains of iRFP713. Requires presence of heme oxygenase for chromophore formation...miRFP709 and miRFP670, pair with common PAS domain. Requires presence of heme oxygenase for chromophore formation...BiFC interaction is typically irreversible and requires some time for chromophore maturation, splitFAST...

...simple, and engineering complex cellular behaviors requires making complex manipulations to cells. For example...number in the Level 2 assembly. These pathways can require a lot of optimization - which is why MoClo is so...of a combination of multiple genes to achieve a desired effect. This is commonly used in plant biology ...

...Systemic therapies involve CRISPR-Cas delivery directly to the patient. Patient-derived cells can alternatively...the goal of ‘fixing’ mutations or introducing a desired edit, but this particular class of treatments has...inflammatory diseases, blindness, and diabetes. Directions and obstacles for future CRISPR therapies CRISPR...

...longer incubation times and didn’t necessarily require cycling. The Pros and Cons Researchers used the...select for correctly assembled phage genomes, it required a very stringent selection process and benefited...selection, failed under normal cycling protocols, and required a 48-hour incubation at 37 °C. While the 52-piece...

...Targeting Receptor Expression Using chemogenetics requires successfully introducing receptors to target cells... to control ion flux. The system allows you to directly manipulate ion channels. When working with PSAM...Magnus C, Lee P, Bonaventura J, Zemla R, Gomez J, Ramirez M, Hu X, Galvan A, Basu J, Michaelides M, Sternson...

...viral infection by bacteriophages), or conjugation (direct transfer from one cell to another). Bacteria need...transfer genetic material to a recipient through direct contact. That’s conjugation! F factors can also...cloning. In addition, scientists have engineered entirely new types of plasmids without clear analogues ...

...active sites. In addition, Casɸ has a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is... is expected to contribute to experiments that require high photostability, such as time-lapse imaging...Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a reduced scale...

...that the orientation of the flanking loxP sites directs the Cre recombinase to invert, translocate, or ... the antisense orientation (Figure 1, top). The entire DNA cassette would be flanked by two pairs of target... Wendling O, Chambon P, and Ghyselinck NB. A directional strategy for monitoring Cre-mediated recombination...

...world. With three Nobel Prize winners, one NIH Director and a co-founder of BGI-China presenting, it’s... discoveries and moving beyond DNA Eric Green, Director of the National Human Genome Research Institute...proteins). The chemistry is complex and this work requires an interdisciplinary team.  Day two of the conference...

...handle or direct with a degree of skill, To work upon or try to alter for a purpose and To direct the professional... these tasks before I was a manager! Second, my direct report was not engaged because I only left him/...