...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...

... capsid of another. Formally, pseudotypes are described with two numbers separated by a slash: the first...through our in-stock viral prep service.  AAV2 Despite being isolated second, AAV2 is the best studied...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...., Robbins, P. D., Li, J., & Xiao, X. (2006). Widespread and Stable Pancreatic Gene Transfer by Adeno-...

...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...

... be spent making and testing entirely new gRNA designs. You can now easily find many validated gRNAs in...validated gRNAs? Proper target selection and gRNA design are essential for a CRISPR experiment.  The selection...headers to sort alphabetically or in ascending or descending order, in the case of numeric columns.  The data...Additional Resources on the Addgene Blog Learn How to Design Your gRNA Use the CRISPR Software Matchmaker Browse...Additional Resources on Addgene.org Browse gRNA Design Tools Read Our CRISPR Guide Pages Find CRISPR Plasmids...

...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...

... 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...

...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,...

...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...

...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!   ...

...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...

...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...

...technical data. These data focus on experimental design and usage of these tools, which aren’t always easy...experiment, use these questions as a guide for designing your future injection experiments. Have your data... exactly where to get that strain. Experimental design & methods: These questions are focused on injection...activate receptors? If you’re in the process of designing your experiment, read our blog post on important...to give proper attribution.  We hope that this provides a clear overview of what you need to submit data...

... 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...

...package. One important concept in FRET experimental design is that the targeting specificity requirements ...codon (often amber i.e. UAG) engineered into a desired site. During translation, the novel tRNA/unnatural...so small, one can insert them in virtually any desired location in a target protein, though predicted ...it all together The FRET donors and acceptors described above can be paired together to measure a wide...combinations of labeling strategies. FRET experimental design lends itself to creativity and innovation, valuable...PMCID: PMC2802762. 2. Dumas, Anaëlle, et al. "Designing logical codon reassignment–Expanding the chemistry...

..., J. C., Kyrpides, N. C., Banfield, J. F., & Doudna, J. A. (2018). Programmed DNA destruction by miniature...hierarchy: class to type to subtype to variant. Despite the seemingly complicated steps to arrive at this...largely underutilized compared to Class 2. Class 1 Despite their incredible abundance, Class 1 CRISPR systems...accessory Cas proteins in the Cascade complex. Despite a tenuous grasp on how type III systems function... III-E effector was developed, called Cas7-11. Despite being in Class 1, Cas7-11 is a single protein effector...