...increasingly able to harness the body’s immune system to fight progressing cancers. It was inspiring to participate...from all fronts!”  Here I aim to compile a few highlights from the conference which reflect how cancer/...targeted engineering of the TCR locus. He also highlighted the UCSF Immunoprofiler - a human biopsy analysis...

...range between 0 and 100) are then converted to a weight (which ranges between 0 and 1) so that they can...ranging from -40 bp to 40 bp) are also converted to a weight (which ranges between 0 and 1) (shown in the graph...Conclusion CrispyCrunch is now publicly-available for high-throughput design and analysis of CRISPR experiments...

...oneself that prompted us to create Crowdfight.   Figure 1: Crowdfight is a community of scientists who...making science.   How we started Crowdfight-COVID19 Crowdfight was born as a collaborative platform...collaborator for a specific task, Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the expert...Ignacio Amigo and María Hernández Sánchez from Crowdfight. In March 2020, while most of the world was locked...did not have access to a P3 laboratory (BSL-3), a high-security facility needed to perform the experiments... them heard about our online platform, called Crowdfight, and filed a request for help. In just a few ... a research group in Scotland, who had just the right infrastructure and offered to collaborate with them...

...hand, there are reasons why grad school might not be the right choice. I spoke with our Financial Operations...here. You also don’t have to go back to school right away. Shannon Rinaldi, Addgene’s Lab Manager, was.... I thought, ‘Let me pursue this, let me make a slight change so I can have flexibility and be up to date...biggest takeaways from my co-op. It has been so insightful to learn about different career paths, successes...

... available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have been ...

...Human Gene Targets The Joung Lab recently used their high-throughput FLASH method to construct a large series...

...Gly-Ser in-frame with prefix start codon; Employs high-efficiency enzymes. Prohibited Restriction Sites...

...and wrap the column with blue LED lights. When you switch off the lights, OptoNBs bind YFP and non-specific... the blue lights. Which option do you choose?  Option #2 may sound like fiction, but light-controlled...dark-induced dark-induced Loop 6 blue-light induced dark-induced blue-light induced Table 1: Summary of the... effect of AsLOV2 insertion site on light-induced binding.  Light controlled protein purification with...while cycling between dark and blue light which revealed a blue light-dependent shift in mCherry fluorescent...cells: light-based control of signaling pathways. This is particularly exciting because using light to control...binding of their target protein is altered by blue light illumination. This ability to reversibly bind and...

...engineering of fusion proteins is relatively straightforward. However, generating a fusion while keeping...discuss strategies to generate fusion proteins and highlight some aspects of their design.  Protein size and...situation Next to size, potential homodimerization and higher order oligomerization of fluorescent proteins is... the fluorescent tag should operate as an inert light emitting module, any tendency of GFP to homodimerize...fluorescent protein characteristics such as the brightness, photostability, maturation time, and acid sensitivity...termini of your protein of interest The most straightforward fusion protein is made by connecting the N-...the fluorescent protein-POI fusions, we generate straight fusions, without any linker. When both orientations...

...the challenge of trying to figure out what system might work best for your particular purposes. We’re here...than others. In today’s post, we will focus on a high-level comparison of popular cloning methodologies...results. If you’re in a hurry, Golden Gate cloning might be for you. This methodology relies on versatile... in as little as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you ...fragments in an order determined by their homology. You might also consider FastCloning, first described by Chaokun...approach should you use if the budget is particularly tight? Most cloning processes will require a PCR reaction... job done, provided that you’ve already got the right enzymes laying around the lab. If not, you’ll be...

...conducted in Professor Cepko's lab and to get some insight into her management and mentoring styles. If you'd...

... focused on three different levels of assembly: High efficiency and accurate single-insert assembly Intermediate...destination plasmid. Antibiotic selection allowed for high throughput cloning efficiency measurements. We performed...confirmed the insertion of the Lambda insert at the same high frequencies as the antibiotic selection markers....lacZ cassette assembly was easily achievable with high levels of transformants and low backgrounds – so...Using these 4 bp overhang observations, accurate “high-fidelity” junction sets for both the 12- and 24-...complex assemblies were performed. It was found that high efficiencies and accurate assembly levels were indeed...in the number of transformants while maintaining high levels of correct assemblies) translate to faster...

... (blue light-inducible AraC dimers in E. coli) that turns expression on and off with blue light.   Inspired...domain with VVD, a light-responsive domain that dimerizes upon exposure to blue light (Fig. 1). Another...VenturaIf you follow Barbara Di Ventura on Twitter, you might have seen the video of her singing about the paper...Another blue light-inducible dimerization domain, Vaucheria frigida aureochrome1 (VfAu1), led to functional...most applications.   Figure 1: With blue light, the VVD domains of the BLADE dimerize and activates...under the pBAD promoter, can be controlled with light without the need of any cloning.     What advantages...BLADE activates transcription with the flick of a light switch and shuts off just as quickly. The original...

..., Chen PW, Casanova JE, Cherfils J, Dacks JB, Lambright DG, Lee FS, Randazzo PA, Santy LC, Schürmann ...

... in bacterial genome editing is not always straightforward. Read our CRISPR Methods for Bacterial Genome...

...human and mouse genes applied to an arrayed viral high-content screen. Moffat J, Grueneberg DA, Yang X,...

... The p53 name arose from its observed molecular weight (53 kDa) when run on an SDS-PAGE gel. Cancer Impact...

...and Cpf1, which require that each DNA target have high sequence identity to the spacer sequence and contain...

...This is where FLEx enters the scene. FLEx: A light when night is about you! The FLEx (for flip-excision)...optogenetics? Optogenetics: Controlling neurons with light In 2005, a team from Stanford University, led by...ago with the discovery of microbial rhodopsins, light-sensitive proteins that absorb photons for energy...maintain membrane potential in response to visible light (Zhang et al., 2011). What do microbial rhodopsins...2000). Most neurons in the brain are not naturally light-sensitive, so the selective expression of opsin ...Optogenetic neuromodulation is made possible by the light: when hit with the correct wavelength, microbial..., 2011; Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in turn, trigger...

...We provide high quality AAV preps from select plasmids in the repository. Browse our control AAV collection...