...to realize that the effect of Dr. Yamanaka's work has been profound, and this is a rare instance in which...address, appeared almost incredulous that the field has exploded so quickly, citing the first clinical trial...scratch the surface of different fields. This meeting has given me an opportunity to dive a little deeper into...scientists care about patients As any grad student who has spent years studying how that one protein interacts...presenters, it was gratifying to see that Addgene has ensured that plasmid acquisition is not a rate-limiting...

...biology. The mistrust and anger directed at GM crops has burdened this emerging engineering technology. Monoculture... metabolism are well understood. Additional work has also identified and characterized mechanisms and ...petunias. Because of the extensive research that has come before us, it’s possible for us to combine these...plant synthetic biology. Plant molecular biology has traditionally relied on overexpression and knockouts...Nikolai Braun grew up outside of Washington D.C. and has always enjoyed exploring the outdoors and experiencing...

...considering my future path. Talking to Addgenies has helped me get a much better idea of what lies ahead...Director of Product Management Eric Perkins, who has been in many roles in Addgene since he started, said... set up for failure. Listening to Kenkel’s story has helped me realize that is not true at all. Racheal...medical school was devastating. At the same time, she has an entirely new path to discover and gives her time...finance, and while I've done that, my big takeaway has been the impact I can have on an organization beyond...was one of the biggest takeaways from my co-op. It has been so insightful to learn about different career...

...technology has revolutionized the fields of molecular biology and bioengineering, as it has facilitated...site, or protospacer, in the genome. Native Cas9 has two catalytic domains, each of which cleaves one .... When native Cas9 is complexed with a gRNA that has been truncated such that it exhibits 15 or fewer ...truncated gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased ...creation of innovative technologies. To date his group has develop several popular CRISPR-based tools for altering...

...positions. There has been a lot of engagement with the Athena SWAN Charter and it has really strengthened...their pathogens. As Patron explains it, her work has always been focused on gene transfer, from transgenes...MoClo) Kit. What does it include? Patron: Sainsbury has a mandate to share and educate and enable plant science...quantitative data for regulatory elements, which has not been available for plants. Addgene: Will the ...strengthened the network of female scientists. Twitter has been really good for that too. I’m more in touch with...

...proteins such as Green Fluorescent Protein (GFP) has likely existed in creatures such as jellyfish for...from the crystal jellyfish Aequorea victoria. It has a fluorescent emission wavelength in the green portion...protein isolated from Discosoma sp. Similar work has been done to expand the red-fluorescent protein repertoire.... Split EGFP: An alternative to FRET, split EGFP has also been used to study protein-protein interactions...A wide array of GFP-based fluorescent biosensors has been designed to detect a variety of intracellular...surface of the potential applications for GFP. It has also been used to identify particular cell populations...

...It’s clear that CRISPR-Cas9 technology has really changed the game for anyone looking to quickly and ...human genes. Unlike the GeCKO library, each library has been enriched for different gene targets. These pooled...system. Genome wide mouse CRISPR library The Yusa lab has leveraged the CRISPR system to target all 19,150 ...stably transfected for Cas9 expression. The Yusa lab has used their library to screen for genes involved in...

...five years, inflation has occurred. Here at Addgene, the consumables cost per stab has increased 50% since...make a plasmid. Doench’s lab in the Broad Institute has created hundreds of thousands of plasmids, using ...since 2019, while overall inflation of USD has gone up 19.8% in the same time period. For ease of calculations...per plasmid, Addgene, since its founding in 2004, has saved the scientific community a collective and very...

...sharing open hardware has a similar goal. The sharing of modules, ideas, and blueprints has the potential not...Since its early years, the light microscope has been a powerful tool in almost all disciplines. In biology...Sharing open-source plans that rely on such tools has allowed the community to regularly replicate designs...portable device for studying plankton using flow cells, has become an established alternative to high-priced ...

... funding model in nuclear receptor research, and has generated a set of comprehensive datasets that collectively...interactome of these key regulatory molecules, and has provided clues to their previously uncharacterized...website. NURSA and Addgene The great progress that has been made in the NR field over the last thirty years...and coregulators, and although the reporter gene has over the years evolved from chloramphenicol acetyltransferase...research tools is a double-edged sword, however, and has greatly increased the burden associated with identifying...

... the best outcomes. The go-to in vivo option AAV has become the preferred delivery method for in vivo ... other viral vector delivery methods. Lentivirus has limited packaging options, with most lentiviral vectors...possibilities when designing screens. Each serotype has its own tropism, or a preference to infect specific...BioRender.com. Beyond the identified serotypes, AAV also has psuedotypes. Psuedotyping is the process of making...Lentiviruses have capacities of 8–12 kb, while AAV only has approximately 4.7 kb — which means that when using...the genetic landscape of tissue-specific diseases has grown exponentially, aided by advancements in in ...

...repository. Our blog is just over a year old, and it has grown steadily during 2014. We were excited to have...continues to grow as the genome editing technique has developed and improved. The CRISPR posts written ...about. Our post on “Making Your Own Competent Cells” has been extremely popular, generating many additional...

... but inadequate access to reagents and equipment has become a bottleneck. In the last few years, scientists...nucleic acids at the atomolar level. The Zhang lab has adapted natural RNase activity of the Cas13 protein...Gootenberg et al., 2017 and 2018). While the Doudna lab has used Cas12a’s non-specific ssDNA degradation to develop...field (Myhrvold et al., 2018). Now, the Zhang lab has shared a protocol for using SHERLOCK to detect SARS-CoV...at atomolar levels, within one hour. DETECTR thus has the ability to rapidly detect nucleic acids with ...diagnostic applications. The company Mammoth Biosciences has been started based on DETECTR technology with the...Biotechnology. Like the SHERLOCK detection system, DETECTR has not been approved for diagnostics yet, but is undergoing...

... applications (v82 has higher affinity for glutamate but slower kinetics; v857 has faster kinetics but...iGluSnFR3 (Aggarwal et al. 2022). This new version has higher signal to noise ratio and is better localized...for two-photon microscopy. The resulting JEDI-2P has faster response kinetics and increased brightness...and deep-tissue imaging. Vladislav Verkhusha’s lab has generated a new genetically encoded calcium indicator...interest, it’s important to choose a biosensor that has a binding affinity close to the range of concentrations...physiology and metabolism under various conditions. LiLac has improved response size and reduced sensitivity to...

...specific protein folding needs. The pCri system has recently been used to study a matrix metalloproteinase... Queensland’s Institute for Molecular Bioscience has provided a solution for this problem by developing...eukaryotic cell extracts (Gagoski et al 2015). He has also constructed a library of eGFP-tagged human ORF...constructs. The Cell-Free Protein Expression Kit has been used to produced recombinant proteins for in-vitro...populations through time, Christopher Marx's lab has engineered and validated a new method called FREQ-Seq...cost-effective manner with minimal bias. The FREQ Seq kit has been used in microbiology studies monitoring sequence...binds your RNA sequence of interest. The Zhao lab has made this easy with their PUF Assembly Kit. The kit...

...nervous system (CNS) (Wu, Asokan, et al., 2006). AAV1 has also been found to transduce skeletal muscle, cardiac... the most common choice for AAV pseudotyping. It has broad tissue tropism, including liver, muscle, lung...generally the best choice for targeting astroglia. It has additionally been reported to have tropism for lung...through our in-stock viral prep service.  AAV8 AAV8 has high transduction efficiency in multiple tissues,...applications targeting liver tissue. In addition, it has been shown to transduce skeletal muscle, cardiac,...through our in-stock viral prep service.  AAV9 AAV9 has relatively broad tissue tropism and high transduction... the CNS (Issa et al., 2023). Additionally, AAV9 has been shown to transduce liver, skeletal muscle, pancreatic...

...excision site after cleavage by the transposase. It has a cargo capacity >100 kB, although integration efficiency...efficiency decreases with cargo size. Sleeping Beauty has a close-to-random integration profile in mammalian...hypersensitivity sites. Like Sleeping Beauty, piggyBac has an integration efficiency in human cells similar ...1996). Unlike Sleeping Beauty and piggyBac, Tol2 has a weak consensus sequence for its preferred target...hypersensitivity sites. Tol2 is only active in vertebrates and has lower efficiency of integration in human cells than... is a truncated version of the original Tol2 and has ~3-fold increase in transposition activity  Tol2...encodes a Type I-F CRISPR-Cas system. This system has four major components: 1) a CRISPR RNA (crRNA), 2...

...graduate program at the University of Manitoba. Our lab has been utilizing chemogenetics (Designer Receptors ...Although AAV2 can transduce many tissue types, it has low expression in several brain regions where other...serotypes to infect different cell types. In the CNS, it has been shown that there is variable transduction of...from the Original Location - Axonal Transport It has been demonstrated that certain serotypes can undergo... of movement, distance traveled, etc.) (4). This has also been identified with AAV2 (5), AAV6 (6) and ...required for the onset of gene expression. However, it has yet to be determined what gene expression looks like...problem in many experiments, a group of researchers has identified this feature of AAVs as a new tool for...

...Research will be held from 22-24 October. This blog has been contributed by guest blogger and Future of Research... of a system that, despite all of its successes, has many deep-rooted flaws (1). As we discovered through...example, Harvard’s Therapeutics Graduate Program has a compulsory internship to be taken in an industry...Marder, professor of Biology at Brandeis University, has recently published an article arguing against reducing...Labor and Worklife Program at Harvard Law School. He has continually promoted the cause of increasing data...

...methods. Each method has its own pluses and minuses, but Golden Gate cloning has been especially useful...number of potential clones. Golden Gate assembly has a few advantages over other cloning methods. Exonuclease-based...just a few steps. More recently, CRISPR technology has adapted Golden Gate cloning for inserting the appropriate...also be adapted to express multiple gRNAs. Addgene has two Golden Gate-based gRNA assembly methods available...