...visualizing promoter activity, and much more. Read on to learn more about GFP, how scientists have evolved this...exist with only slight variations separating them.  Please note that many fluorescent proteins found on the...F64L; S65T, Y66W; N146I; M153T; V163A; A206K Cerulean F64L, S65T, Y66W, S72A, Y145A, H148D, N149I, M153T... Y145A and H148D Stabilizes the structure of Cerulean derivatives F99S, M153T, V163A Improves folding...visualize the effects of alterations to this activity. Learn more about optogenetics at OpenOptogenetics and ...

...different chromatin states. Alexander Stark and colleagues at the Research Institute of Molecular Pathology...Initiative, led by Dominic Esposito, has recently released a collection of 360 Ras pathway plasmids to the...Drosophila cells, targets 13,501 fly genes with at least three independent gRNAs, each chosen strategically...vacuolar) pH environments that inactivate SEP and leave only DsRed to fluoresce (1). With variants that ...effects on gene regulation. Professor Zoldos and colleagues from the Department of the Biology, Division ...26912792 FusX-TALEN assembly system TALEs as nucleases (TALENs) are genome-editing tools that are deployed...auxotrophies or using different auxotrophic backgrounds can lead to disparate physiological effects and complicate...

...integrating neuronal signalling and triggering release of the stress hormone norepinephrine to promote...are chemogenetically activated, norepinephrine release causes a freezing response, a form of immobility... the NAc, where enhanced glutamate transmission leads to locomotion sensitization. (for review see van... as sleep, food intake, mobility/coordination, learning, and memory. We’ll cover these in a  future blog...Guide to Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...tips from an AAV rookie  Learn about some considerations when using AAVs Learn more about using AAV as ...complex (LY6A) as a receptor for PHP.B. Read on to learn how the design of PHP.B may have contributed to ...182-188. PubMed PMID: 29175632. If you want to learn more about how PHP.B and PHP.eB were engineered,...PMC5088052. Additional resources on the Addgene blog Learn more about PHP.eB, and PHP.S, a peripheral nervous...

...CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear expression of BFP2.     Figure 1: CROPseq-multi...separated multiple sgRNAs with tRNAs, which are cleaved out of the RNA transcript by endogenous processing...and found the protein La (a small RNA-binding exonuclease protection factor) promotes editing (Yan, J et...hypothesized that the improved efficiency is at least in part due to La binding and stabilizing the 3’...Neurodegeneration Collection updates Last but not least, check out the new and noteworthy tools featured...

...each viral lot. To learn which specific QC experiments were performed on your lot, please contact us. Full...medium (OptiPro SFM) containing lentivirus is first cleared by low-speed centrifugation and then by filtration...responsive element (RRE) gene and the cellular ribonuclease P/MRP 30 kDa subunit (RPP30) as a control for...the sample from the DNA tube all the way to the cleared viral preparation pool. In addition, the final ...

...replicon of pTargetF may exhibit a more serious leaky expression in BL21(DE3), which we presume caused...that indeed the gRNA-pMB1 had a slightly higher leaky expression in BL21(DE3) that resulted in a lower...a sacB gene renders it beneficial for plasmid clearance. It is more convenient to update the spacer(20nt...Anhydrotetracycline (aTc) induces expression transposases leading to increased integration frequency. (C) Increasing...rounds of transfer on transposition-inducing agar lead to higher cargo copy numbers. (D) The effect of ...

...Blog Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian Cells Learn about...older than those available from biorepositories leading to genotypic and phenotypic artefacts. The life...commercial repositories and the same cell line from a colleague?”. The main difference is the QC and validation...usually the case if you obtain a cell line from a colleague. Time and money are wasted, papers are retracted...

...remains unclear whether the FP misfolding impacts fusion protein function and this could lead to under...localize [4]. Most investigators are surprised to learn that FPs are often poorly suited for cellular compartments...glycosylation, disulfide bond formation, and proteolytic cleavage. FPs localized to organelles are equally susceptible... dark fluorescent proteins in your experiments, please consider the following: How can one ensure that...

...bit of a steep learning curve and many opportunities for mistakes. The easiest way to learn optogenetics... is an approximation of reward or the sense of pleasure that the animal perceives through taking a drug...University.  His interests include the role of learning and memory in opioid addiction and science communication...Kheirbek, Mazen A., et al. "Differential control of learning and anxiety along the dorsoventral axis of the...

...Bench series was born from the shift to remote learning due to COVID-19. We had many protocols for plasmid...details about personal protective equipment to learning about how to use a tissue culture hood and other...other lab equipment, we’re here for you as you learn the ropes! All material can be found on our Protocols...Protocols page and YouTube page.  Happy learning, and welcome to the lab bench, new scientists!   ...

...investigating HIV integrase mutants in the early 1990s (Leavitt et al., 1996), and since then, numerous variants...However, IDLV genomes are transcriptionally active, leading to transient expression of gene products (Wu et... insertional mutagenesis. This integration could lead to an undesirable interruption of a critical gene... off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology, 31(9), 822...PMID: 18688286 PubMed Central PMCID: PMC2496964 Leavitt, A. D., Robles, G., Alesandro, N., & Varmus, H....

...warm meal on a winter day, you should be happy to learn that many DNA-based reagents are named for soup ... and non toxic to cells. Because it rapidly photobleaches, DFHBI can only be used for qualitative, but...regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that control the expression...the Corn-DFHO complex. For dessert (last but not least), the only fruit-named aptamer, Mango, binds to ...PMC4143336. Additional resources on the Addgene blog Learn more about aptamer fluorophores Investigate other...

...the uses of biotinylation in molecular biology! Learn about in vivo biotinylation of bacterial fusion ...biotin-avidin complexes bound to your protein of interest leading to amplification of the reporter signal and easier...microenvironments. There are also reversible and cleavable biotinylation reagents to help with the specific...vivo biotinylation of bacterial fusion proteins Learn how biotin is used in Split-BioID: An improved method...method for studying protein-protein interactions Learn how biotin can be used with CRISPR Additional resources...

...expressed proteins endA, endA1 Endonuclease I (nonspecific cleavage of dsDNA ) mutation Improves plasmid...cell mcrA, mcrBC Inactivation of pathway that cleaves methylated cytosine DNA Allows for uptake of foreign... mrr, Δ(mcrC-mrr) Inactivation of pathway that cleaves methylated adenine or cytosine DNA Allows for uptake...recombination, increases plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability...

...blog posts Learn more about AAVs Resources on the Addgene website Visit the AAV Data Hub Learn more about... serotype, injection site, and incubation time. Learn more about the Data Hub in this blog post. Alongside...attending SfN2019, stop by booth #2163 to say hi and learn more about Addgene’s viral service. Additional ...

... the Addgene Blog Learn How Piggy Bac Vectors Can Be Used for Genome Editing Learn about Michael Koeris...needs. For more context, exposition, and detail, please refer to the paper. Why might one be interested...uploaded to Addgene. We also recommend inserting self-cleaving hammerhead ribozymes directly upstream of the ... transcribed gene and its 5’ UTR. Various self-cleaving hammerhead ribozyme parts are described in Nielsen...

... I had found my dream job, because one thing I learned in grad school was that scientists have amazing...at their lab benches. It’s these exchanges with leading scientists (and our growing collection of 25,000...sciences research for the past decade. We’ve also learned along the way that a connected and collaborative...scientists who have deposited plasmids at Addgene.  Learn how discoveries were made and get practical tips...

...Health and Allied Sciences in Ghana. Read on to learn more about doctors Bengyella and Duedu and how they...tumors is “a major public health problem, and a leading cause of hospitalization for gynaecologic disorders...Program Additional Resources on the Addgene Blog Learn about Plasmid Awards to Researchers Working on Rare... in China Additional Resources on Addgene.org Learn About Addgene's Mission Check Out Our International...

...even to not bind to their target molecule at all, leading to inconclusive results, erroneous conclusions,... that your antibody has been validated using at least one of these strategies. Choosing which strategy...) the protein of interest in your sample, should lead to reduced or eliminated antibody signal. If you...translation start sites or alternative splicing that could lead to some form of the target protein to be present... Check out our Selecting an Antibody blog post! Learn more about Antibodies and their many applications...

...alcohol-based disinfectants. A 10% bleach solution should be used to clean AAVs from workspaces. Speed...Manitoba due to their Biosafety Level-2 or 2+. Please Note: Despite the fact that AAVs themselves are...vectors (e.g. Lentivirus, Adenovirus) AAVs cause the least amount of immune response. Any investigator who ...antisense oligonucleotides (ASOs), zinc finger nucleases, and many others (12). The most attractive potential...

...LTRs) has been removed, Rev (which facilitates nuclear export) is expressed from a separate plasmid, and...mutation or recombination with human retroviruses could lead to RCL. Regarding transduction efficacy, no differences...Quickly after transfection (4-8h), cells start releasing viral particles into the culture supernatant and...retrovirus packaging cell lines to avoid recombination leading to helper virus production." Molecular and Cellular...5 Tips for Trouble Shooting Viral Transductions Learn Why You Might Want to Use AAV in Your Research Lentiviral...

...countries. We, like everyone around the world, are learning to navigate the changes brought about by the pandemic...opportunities. I have spent the last 9 months quietly learning about Addgene’s past and present, but the real...successes to reflect upon, a wealth of history to learn from, and a compelling future to plan in the changing...people. I look forward to working with Addgene colleagues, collaborators, partners, and the larger scientific...

... change? Addgene has you covered. Today we’re releasing 11 bold science-themed backgrounds! Show off your...out Addgene’s Fluorescent Proteins blog topic to learn more about FRET and other techniques using FPs! ...Addgene are shipped as bacterial stabs? You can learn about what format your material will come as (and...molecular biology tools in your research or not! And learn more about the variety of careers available to scientists...

... guide Learn about different careers in science communication Resources on Addgene.org Learn how Addgene...main three points of your story. Interested in learning more? Visit their website at www.theaddictivebrain.com.... Sadangi is an eLife ambassador, SfN community leader, and an early research career panelist at BioTechniques...

...Resources at Harvard Medical School, Joanna was my colleague. She mentioned Addgene was looking for someone...jewelry making, reading, sailing, and gardening. Learn more about Julie Cicalese  Ms. Cicalese served as... for Harvard Medical School where she brought leadership teams together to advance the mission of the ...England. Additional resources on the Addgene blog Learn more about about Addgene in our "Inside Addgene"...

...plasmid from 2016 is an expression vector for CRISPR nuclease, SpCas9 from the Zhang Lab. It's name, though ...with the plasmid post transfection. The 2A "self cleaving" peptide causes the ribosome to translate Cas9...gRNA that will target SpCas9 to the approrpriate cleavage site). Finally, in addition to the 2A peptide ...Resources on the Addgene Blog Browse Our CRISPR Posts Learn about Using CRISPR to Generate Knockouts Read Some...

...applications was still very new. Now, Konermann and her colleagues including Zhang and Mark Brigham have taken the...influences in our environments. We are able to react and learn. All of this requires genes to be regulated dynamically...modifications in the genome,” Zhang said in MIT’s press release. In an accompanying Nature News & Views Andreas...processes and disease states. While LITE plasmids have clear application to anyone with an interest in neuroscience...

... well-integrated into our lab lives. Read on to learn about some creative approaches to lab sustainability...Who even regulates them? You may be surprised to learn that it hasn’t even been 50 years since the creation...fluidic machines handle biological samples and are cleaned between each use. You wouldn’t throw away your ...series!   Additional resources on the Addgene blog Learn about recycling initiatives for the lab Subscribe...

...King soundtrack and writing this post, it is my pleasure to announce that we once again reached new heights...section to make this post even more useful to your colleagues. Technique: Probe Phage Genomes for Host Binding...(check out our interview with Michael Koeris to learn how his company uses phage to test for Listeria ...posts in chronological order here: Four Things I Learned at the 10 Years of iPSCs Symposium Tips for Writing...