...until 2005 [1]. Like other transposons, piggyBac™ has two components, a transposon and a transposase. The...offers a large cargo-carrying capacity (over 200 kb has been demonstrated) with no known upper limit [1]....particularly useful when the gene being expressed has threshold effects, such as with signaling molecules...transgenic cell line with the piggyBac™ system is that it has a preference for integration at regions of open chromatin...removal, which alters the endogenous DNA sequence and has been suggested to cause cytotoxicity [5]. Second,...efficiency. While the efficiency of excision with PBx has been shown to be higher than that of the wildtype...Cre recombinase, the PBx is highly specific, and has not been shown to cause genomic abnormalities when...

...mutagenesis or gene targeting Victor de Lorenzo's lab has engineered a modular mini-Tn5 vector that can be ...and the subsequent biochemical characterization, has enabled scientists to engineer a wide variety of ...green to red when it is oxidized. The Timer tool has already been used by several groups to monitor protein...imported to the mitochondrial matrix. Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-... for protein-peptide locking, Mark Howarth’s lab has developed a new tool for peptide-peptide locking....optimization through engineering, Ed Boyden’s lab has identified a variety of new channelrhodopsins for...nicknamed Chronos, is blue and green light-excitable and has faster kinetics than those of other channelrhodopsins...

...restriction enzyme, you will need to verify that it has be cloned in the correct orientation - this can be...

...Yeast Extract and dH 2 O to 400 mL Note: If your lab has pre-mixed LB agar powder, use the suggested amount...

...between a cell that has had one viral particle integrated into its genome and a cell that has had multiple integration...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected... be significantly higher than the same prep that has been stored in the freezer. If you plan to freeze..., you may want to consider using an aliquot that has been subjected to a freeze/thaw cycle for titering...CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used for Lineage Tracing Additional Resources...

...science has become increasingly restricted to academic and industrial labs, but recently there has been ...KittyBiome has been incorporated as a non-profit entity and so far most of their funding has come from ... the predictions about the effects this mutation has on locomotion in our computer model to the effects...Genetics Center (CGC) at the University of Minnesota has declared our makerspace an educational institution...

...to define how you will know that the relationship has been successful – what are the desired outcomes? ... Dora Farkas, founder of Finish Your Thesis (who has also blogged for us) offers a free report on her ...called “Secrets to Success in Graduate School”. She has also launched an online career coaching program that...in Madison, Wisconsin she also works remotely and has experience as a scientific recruiter. Chris Humphrey...of From PhD to Life. She runs great tweet chats, has an outstanding blog and, while her PhD is in History...remotely, will travel for speaking engagements and has many areas of specialty from leadership development...

...also the fastest test that has been developed so far.  A similar method has been optimized to detect the...Two-step-SHERLOCK has a sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in...State University. Since its appearance, SARS-CoV-2 has spread to almost every part of the world manifesting...full-fledged pandemic. Containing the spread of this virus has become an utmost priority for countries around the...the outbreak ultimately flattening the curve, it has become evident that widespread testing is crucial... 83 known COVID-positive samples tested, DETECTR has a positive prediction rate of 95% and a 100% negative...

...in living cells. The HA frankenbody is an HA tag detection probe. It contains an HA-binding scFv that’s...often genetically fused to other proteins, which has led to their application for basic and translational...fluorescence intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged...s fused to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based probe...of cytotoxic granules from the T cell. When a CAR has an scFv that binds cancer-related antigens, binding... Addgene blog: Read about frankenbodies, an anti-HA scFv-based probe for fluorescent imaging. Learn more...

... dark but dissociate under blue light. This tool has now been modified by Torsten Wittmann’s lab at UCSF...genetic or pharmacological manipulations on cells, and has potential use for a wide range of applications in...monomeric, 4-5 fold brighter than mNeptune684, and has a favorable pKa of 5.6. To test out this new FP in...The increasing popularity of single base-editors has resulted in the rapid expansion of the genome editing...expansion of genome editing tools, the Reuben Harris lab has deposited a reporter plasmid that allows for rapid...

...There can be no doubt that CRISPR/Cas9 technology has been a breakthrough for the genome-editing field ...CRISPR Therapies Cancer and CAR-T therapy The FDA has already approved CAR-T immunotherapy for cancer treatment...CRISPR Therapeutics. Data shared from these trials has been outstanding, with almost all patients experiencing...desired edit, but this particular class of treatments has quite the opposite aim: to destroy. One treatment...way. Delivery, especially for systemic approaches, has its own unique set of obstacles. Thus far, several...delivery tends to be trafficked to the liver. Thus, it has been used for hATTR therapy, since TTR is primarily...

... It was inspiring to hear how the genome project has already had a major impact on undiagnosed and rare...its work on the origin of cellular life. His lab has long hypothesized that life started with RNA and ...prokaryotic genome. Then he desribed how P. furiosus has systems to defend against both DNA and RNA viruses...technical developments. Using the Tet-on system, his lab has created a tetracycline cas9 inducible knock-out system...in a complicated conformation “not like DNA which has a boring structure.” That certainly summed up this...this conference—nucleic acid science still has many surprises for the researchers in this field. Resources...

...enormously plastic tool and has taken the scientific world by storm. While Cas9 has been most widely used to... to create specific edits in DNA, there has also been significant work on constructing Cas9 transcriptional...CRISPR activation for genes with known functions has the potential to be used in therapeutics, as it can...changes in phenotypes. For example, CRISPR activation has been used to differentiate induced pluripotent stem...creation of innovative technologies. To date his group has develop several popular CRISPR-based tools for altering...Genome-wide Screens with CRISPR/Cas9 Learn How CRISPR Has Been Used with AAV Resources on Addgene.org Find...

...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...

...Recently, Xue Sherry Gao’s lab at Rice University has developed DAP arrays for efficient multiplex base...process and release its own gRNAs from a single array, has been a top choice for multiplex gene knockout or ...proper functions to enable efficient base editing and has yet to be developed for prime editing, both and their..., which is conserved in all living organisms and has previously been used for multiplex Cas9-nuclease ...lengthy promoters, such as U6 or EF1α promoters, has limited the number of gRNAs that can be packaged ...

...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...