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...researchers at Japan’s Hiroshima University who described their new tool in Scientific Reports in June. ... gap. As Sakuma and colleague Takashi Yamamoto describe in their recent study, the construction system...vector-mediated multiplex genome engineering. The system now provides an efficient targeting strategy for multiplex ... validated and ready. “Currently, the kit only includes two versions of Cas9, nuclease and nickase, but...Addgene here. Other Popular CRISPR Posts: How to Design Your gRNA for CRISPR Genome Editing CRISPR-Cas9...

...antibody worked under the conditions described above, and a short description of how the antibody performed....the submitted application, along with a brief description of the experiment. If that data was included ...ELISA — but in all cases, it contains a detailed description of the experimental conditions.     Figure 4...The submission process is easy, with simple, descriptive fields, drop-down boxes when appropriate, and...

... capsid of another. Formally, pseudotypes are described with two numbers separated by a slash: the first...cardiac, and retinal cells (Issa et al., 2023). AAV2 Despite being isolated second, AAV2 is the best studied... choice of AAV vector inherently depends on the design of your experiment. Though we cannot make universal...experimental reports on commonly used vector cargo designs and administration doses. Don't forget to submit...2008). Adeno-Associated Virus (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1,...., Schaffer, D. V., & Karpova, A. Y. (2016). A Designer AAV Variant Permits Efficient Retrograde Access...

...In 2005, Boyden et al. described the first light-activated tool for controlling neuronal activity, channelrhodopsin... light in combination with retinal) variations designed to give researchers flexibility in using optogenetic...precise timing of the perturbation. Other tools are designed to be activated within a narrower band of wavelengths...function through multiple intra-cellular signaling cascades. Optical methods for controlling GPCR activation...control GPCR activation and downstream signaling cascades (reviewed in Abreu & Levitz, 2020), including ...You’ll need to test for these differences when designing your experiment.  Finding the right information...

...colleagues describe in the PLoS ONE paper that introduced the kit in December, GreenGate includes the six... bring this into your target organism. We have designed a Golden Gate system based on the daily needs ...and cut another,” Lohmann said. “It allows us to design modules that always have the same recognition site...produced will have different overhangs, they can be designed such that they go back together only one way and...

...record breaking month and to find other posts that deserve a second look. 5 Great Apps for Lab Life This very...Stacey from Eastern Tennessee State University and provides a brief rundown of Sean’s top five smartphone ...Jessica Sacher from the University of Alberta and describes a technique you can use to find phage proteins...molecular biology tool, P1 phage. 2) The post describes an ingenious but simple technique to screen for...that, while affecting a small number of people, deserves our attention and has huge potential for the development...

...so he assembled a plasmid kit designed to do exactly that. As described in the journal Science Signaling...When cancers are treated with drugs designed to hit them right where it hurts, the effects are often ...mutant complementary DNAs (cDNAs) representing core nodes that, when over-expressed, constitutively activated...activators for more cancer-related pathways and more nodes within those pathways. With support from the National...

... discusses some of the programs SITN runs and describes some of the more popular topics from the Science... fields as well as students from the ed school, design school, and Kennedy school to discuss the impacts...impacts of science in a social context. Episodes are driven by narrative and story telling. Chapter 5: The...The SITN Blog - Vini and Amy describe how the SITN blog is a source of scientific information for the ...underwater? Chapter 8: Work-Life Balance - Vini describes how she balances working in the lab and working...

...experience. Most Addgene PhD level job descriptions clearly describe that the roles do not involve bench ...submission of a job application. 3) Read job descriptions carefully Be open minded and be bold. You don...qualifications to apply for a position.  A job description is usually a wish list and the hiring manager... way to a hiring manager. Learn to confidently describe your strengths in your resume and don't be shy...

...eukaryotic process where approximately 20-30 nucleotides of double-stranded RNA (dsRNA) results in downregulation... of an RNAi-inducing transgene. Typically, the desired target of RNAi is expressed as inverted repeats... and quantify actively silencing plants. Non-destructive methods, most popularly with a fluorescent reporter...Fluorescent reporter-based RNAi constructs have been designed and widely implemented in plants, such as in the...approach, myself and others in the Luis Vidali lab designed a new RNAi method. Instead of using a fluorescent...functional APT will convert 2-FA to cytotoxic nucleotides, resulting in death. In the presence of 2-FA,...

... sequence that moves around. This DNA sequence encodes the proteins the transposon needs for transposition...contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase and integrase enzymes...ORFs) which often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase activities...preferred target site for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from...encode reporter genes, mutagenic cassettes, or barcodes. When transposons are delivered to cells or a ...occurring Tn7 transposon found in Vibrio cholerae that encodes a Type I-F CRISPR-Cas system. This system has four...

... antibody sequences, degenerate primers can be designed to selectively mutate specific regions of your...sections. Structure-guided directed evolution If you desire a bit more control, structure guided directed evolution...complex as a template. This structural framework then guides the specific mutations you choose, whether it's...allows you to combine the strengths of rational design with the power of directed evolution, all while...technologies, the Institute of Protein Innovation (IPI) provides information on their large-scale recombinant antibody...synthetic antibody products and development methods. Despite recent advances, there is still always room to ...

...teaches you how to design a course, how to design good lesson plans, how to design test questions, and...and how to design lectures. I really built a course from the ground up.”  Science writing and editing These... was for Cactus Communications, a company that provides scientific editing services. “I was working during...

...plasmid The packaging plasmid aka the RepCap plasmid encodes two genes: the replication (rep) gene and the capsid....com   Replication (rep) gene The rep gene encodes four proteins that are required for viral genome...cross-packaging.  Capsid (cap) gene The cap gene encodes three AAV viral capsid proteins VP1, VP2, and VP3...type tropism. Introducing changes and/or short peptides sequences into these variable loops is one method...method for changing a capsid’s tropism.  cap also encodes the Assembly-Activating Protein (AAP) and the recently...

...You’ve designed the perfect experiment – controls, conditions, and everything in between – now all you...This is where affinity tags come in – they are peptides which do not naturally exist in most expression...protein for downstream applications Below, we describe a few of the most common tags available. Maltose...antibody is an alternative to amylose resin and also provides a visual detection mechanism via applications ...especially if downstream antibody detection is desired… and yes, we have an antibody for that!   ...

...problem - see our recap below! Day 1: gRNA design tools: How to Design your gRNA for CRISPR Genome Editing and...given our CRISPR Plasmids and Resources a fresh new design, sorting the plasmids we have available by category...capabilities. Epigenetics: CRISPR has made it possible to design programmable epigenome editing tools, and the field...cleavage in bacteria. In mammalian systems, Cas13a degrades only the specified RNA target. Bonus: It displays...

...fly fishing, and local grizzly bear attacks. He described one such recent attack as "hilarious". Welcome... Mehmet Fatih Bolukbasi from Scot Wolfe's lab described a chimeric Cas9 that is improving the nuclease... and foremost, from the custom software used to design complex genetic circuitry to the downstream output...each, and Tim Lu even compared his "memory" gene codes to a Turing machine (also discussed in the book)...also got to show off our new CRISPR page (which describes how to plan your CRISPR experiment for our new...

...smaller gel apparatus until I settled on using glass slides. With just the right amount of agarose, I could...right amount so the agarose doesn’t pour over the sides.  These tiny gels and tiny lanes required less agar...satisfying?  With practice, I got good at this despite the fact that my lab mates thought this was hilarious...minimize the required area and buffer volume.  So, I designed my own gel box (see illustration above) and bribed...plastic and solder this thing together.  One of the downsides—there was no cover so you had to be careful not...

... Antibiotic selection would only then kill the desired plants! PCR or fluorescent markers are currently... have similar drawbacks, often requiring the destruction of plants or tissue for GUS staining or are not...from heterozygous individuals. Only 1/4 of the descendants of the heterozygous plants, therefore, are useful...from these plants will be green, containing the desired mutation but lacking both RUBY and the construct...compared to other markers lends itself well to nondestructive selection. If RUBY was integrated into a CRISPR...

...our AAV preparations. We recently published a description of the VGS process and use cases in Human Gene...ligation and tagmentation preparation approaches described previously require a preliminary double stranding... the expression cassette that we want, it also includes a small amount of spurious DNA from the packaging...plasmids. We decided to use this to our advantage by designing a Python script that looks for pre-defined serotype...signature sequences in the data. Unlike previously described thermodynamics-based methods of serotype confirmation...