...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...

...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...

...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...

...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...

...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...

...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...Natronobacterium gregoryi, or NgAgo for short. Addgene has already distributed this plasmid all over the world...Publication at Addgene Cas9 and NgAgo head to head NgAgo has a number of unique features compared to Cas9. These...compare CRISPR and NgAgo, but it seems that NgAgo has lower potential for off-target effects. In Gao et...

...2015 has been an exciting year for the Addgene blog. We’ve released over 80 posts on a diverse array ...Scientists and the Plasmids 101 2nd ed, the 3rd edition has now been released), and have started providing recorded...incredibly happy to see that one of our career posts has made it to the top of the list this year. While we...doing just that. Do you like the direction the blog has taken over the past year? Do you think there are ...

...dual-nuclease Cas9-Cas9 chimeras Scot Wolfe’s lab has developed dual-nuclease Cas9-Cas9 chimeras with higher...been many Cas nucleases developed for plants, it has been difficult to compare the performance of different...recently discovered in 2017. Now Dominik Niopek’s lab has adapted anti-CRISPR proteins for optogenetic control...

... disease once limited to small pockets in Africa has spread to urban areas and across the ocean. A nurse... considered unlikely to change – but the illness has been fatal in 70 percent of the most recent cases... in the New York Times and on his blog, where he has offered to answer readers' questions. If you have...

... and software development has caught up with it. The peer review process has been challenged with many...targetable NGG PAM sites whereas the human genome has about 300 million. With such a sparsely targetable...needs of each individual; the  community as a whole has yet to settle on any true kind of value. I’ve dubbed... genome editing tool, the CRISPR/Cas9 technology has been surrounded by a whirlwind of research activity...significant improvement. As a result, the software piñata has been filled with slight variations on a theme without...evaluate off-targeting, but currently no software has been designed for this purpose. This second area ...findings to help everyone else. CRISPR technology has reached into so many different communities that it...

...certain experimental setups. The brightness of NLuc has caught the attention of researchers wanting to perform...proteins can be excited by Nluc emission and the lab has dubbed these fusions LumiFluor Reporters. The LumiFluor...mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness and improved maturation...with Luciferases can overcome these problems but has been hampered by dim signals from the coupled luciferase...ReNL; plasmid #85203) NLuc fusions. The Nagai lab has additionally deposited more than 25 fusions of the...Pairing NanoLuc® luciferase with fluorescent proteins has allowed researchers to make innovative tools not ... Technical Services Scientist, Kyle Hooper, PhD, has also worked with R&D for product development and ...