...than 75% photobleaching after just 1 minute of exposure. MG-2I-dL5** experienced minor bleaching after longer...ablate cells in culture and in vivo.  Read on to learn more about this killer illumination technique! ...real time.  In both cases, dL5** was tagged with mCerulean3 (mCer3) for visualization and expressed from ...Sign Up to Receive the Fluorescent Protein eBook Learn about Small Molecule Fluorophores Resources on ...

... of the mRNA, leaving only a red FP bound to the reporter. Because it contains a nuclear localization ...cell. When cellular mRNAs are digested with ribonucleases, ribosomes protect the segments of mRNA to which...occur. For a TRICK reporter mRNA this displacement leads to a change in the fluorescent signal: untranslated...Addgene. Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background...

...updates to iGluSnFR and SF-iGluSnFR have made it clear that it’s time to update our iGluSnFR post! Here...give researchers the ability to monitor glutamate release from neurons and other brain cells in vivo. Glutamate...protein binds to glutamate, conformational changes lead to an increase in fluorescence intensity from the...differentiating between synaptic vs. extrasynaptic glutamate release. Building on the previous generation of the SF-...Resources Additional Resources on the Addgene Blog Learn about other Fluorescent Biosensors A Practical Approach...

...incorporation of a 3’-5’ exonuclease in the reaction mixture. The exonuclease removes mismatched 3’ overhangs...guide the joining reaction. BioBrick Assembly will leave scar regions at the site of fragment joining, but...still creating scarless plasmid inserts.  Want to Learn More about DNA Assembly? Check Out Our Plasmids ...      Additional Resources on the Addgene Blog Learn More about Gateway Cloning Gibson Assembly and Other...

...complex processes can’t be overstated, we can also learn a lot from Nature’s simpler solutions to transmitting..., and they can rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more...appealing target for bioengineering, but it wasn’t clear whether they could 1) be decorated with proteins...Tunable Protein Piston That Breaks Membranes to Release Encapsulated Cargo. ACS Synth. Biol. (2016). doi...

...libraries Learn about other members of the Addgene Advisory Board Additional Resources Learn a little...post-doc, Doench worked on using RNAi screens to learn more about cancer biology in Ed Harlow’s group at...and “with CRISPR, just the results are so much cleaner. The time frame from doing your primary screen ...Institute here Read other blog posts about CRISPR Learn more about the Broad Institute and the Platform ...

...Addgene’s blog to learn about making your own competent cells and our protocols page to learn about bacterial...have studied how HGT occurs in nature and have learned how to introduce genetic materials into cells in... and can include plasmid DNA or fragmented DNA released into the environment. Various factors promote ...more viral particles. Eventually the cell lyses releasing the newly formed viral particles that can infect...

...illumination. This ability to reversibly bind and release a protein of interest in response to light allows...reaches the nucleus where gene expression changes lead to cell proliferation and differentiation. The lab...739201.  Additional resources on the Addgene blog Learn about using nanobodies to control protein activity...Read about RANbodies, reporter nanobody fusions Learn more about using the secondary nanobody toolbox ...Addgene.org Find nanobody expression plasmids here Learn more about optogenetics in Addgene's optogenetics...

...a team that created a novel form of AI that can learn new tasks and beat humans at the ancient Chinese... side you’re sure to have many opportunities to learn about and develop technologies that will have impacts...pulled tools from across the biological spectrum to learn more about the brain and neurological disease. I...better understand how aberrations in brain biology lead to altered function or maybe your favorite FRET-...pathways. Many scientists came to the Addgene booth to learn more about these tools and to tell us how useful...

...rapid multi-part DNA assembly method. Read on to learn how to use it in your research! The starting point...The key is T4 DNA polymerase, which has 3′→5′ exonuclease activity. T4’s polymerase activity can be thwarted...available to polymerize, allowing the slower exonuclease activity to proceed. By including only one type...researcher can decide exactly how far the T4 exonuclease proceeds: at the first occurrence of the chosen...overhangs must not contain the dNTP present in the exonuclease reaction or polymerization will occur at that...

...the following information for a Cas enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement... (one of the most frequently used Cas enzymes) cleaves both strands of its dsDNA target in cis and has...generates staggered DNA nicks at its target in cis and cleaves ssDNA in trans. In sum, CasPEDIA documents the ...along with its action on the substrate (nick, bind, cleave, mixed). This information represents the ‘big picture...Perhaps you are a structure junky who just wants to learn about Cas domains. CasPEDIA either directly has ...

...inspired you to perform this analysis. Having a clearly stated, specific goal for each analysis will help... currently working in) every time you use them, leave them in their own folder. Doing so will ensure that...same directory where it was run. This technique leaves a trail that further helps you keep track of your... entered is useful for beginners who are still learning the basics. However, these notes take up a lot...Read Our Tips to Keep Your Lab Notebook Organized Learn about Software You Can Use to Save Time and Track...

... a date or two for beer, coffee or meals with colleagues you know will attend the conference. Invite others...sure your name tag is always visible. Some people learn names by seeing them. It is OK to ask someone to...the same talk. Read more on this advice to “never leave an empty seat” here. If there are communal meals...especially if they are not in academia. Make sure to learn how to use LinkedIn properly. Then, follow LinkedIn... lunch twice turns a stranger into a friendly colleague. Recommended Reading & More: Scientist Networking...

...will never ever stop planning, perfecting, and learning, but there are a lot of things we actually don...much potential for service and contribution! Leadership On an organizational level, we keep passing...service and support. The Addgene management team leads with excellence. They are one of the main reasons...the Addgene blog Read our viral vector blog posts Learn more about Addgene Resources on the Addgene website...website Find plasmids for your research  Learn about our viral service Check out job openings at Addgene...

...conclusion of inter-homologue recombination was released. This blog has not been updated in response to...to understand what Shoukhrat Mitalipov and his colleagues have achieved, and how the field will move forward...correcting an autosomal dominant mutation in MYBPC3 that leads to hypertrophic cardiomyopathy. An adult male with... appealing for heterozygous mutations, it will clearly be much more challenging for homozygous mutations...editing rates vary widely from gRNA to gRNA, it’s unclear where this study would fall in terms of editing...

...this gap by both highlighting that researchers do learn MANY transferable skills while working in the lab...a report to communicate your findings to your colleagues. So perhaps a better definition of “problem solving...challenges we encounter. As you spend time in a lab learning techniques and taking on independent projects,...opportunities to practice problem solving skills and learn to apply them flexibly in a range of contexts. As... organization Time management Managing a team Leadership Resources on Addgene.org Check out our careers...

...ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll find in this ...Caenorhabditis elegans on extrachromosomal arrays can lead to artifacts due to overexpression. CRISPR can insert...Ingaramo, Jerome Goudeau, Cynthia Kenyon, and colleagues at Calico Life Sciences. Like many split FPs,.... To resolve this issue, Shangqin Guo and her colleagues at Yale University fused a color-changing fluorescent...codon-optimized, engineered Streptococcus canis Cas9 nuclease that has a broad 5′-NNG-3′ PAM. Get the plasmids...

...possibly be made up of. The answer is… drum roll please… non-coding RNAs! You probably didn’t see that ...activation (CRISPRa) are powerful tools. CRISPRi uses a nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily...targeting ncRNAs. For example, the Cas12a variant can cleave single-stranded RNA using gRNAs to specifically...Charpentier E. A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012...induced by CRISPR/Cas9 in the 5’ region of microRNA lead to its depletion and Drosha processing retardance...

...vector via Golden Gate cloning, perform an exonuclease cleanup, transform, plate, and pick. In short, the...found instead ones that were… pretty close, or at least close enough, you’ve found yourself with a common...Fragmid uses Bbsl, a Type II restriction enzyme that cleaves outside of its recognition sequence. This means...in duplicate, they were able to confirm that at least one plasmid was the correct sequence for every individual...

...Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess...variants By Mike Lacy In 2022, Atsushi Miyawaki and colleagues identified a highly bright and stable fluorescent...just a few days later, Mohan Balasubramanian and colleagues showed the single mutation E138D is sufficient...is worth mentioning that Kiryl Piatkevich and colleagues generated yet another monomeric StayGold variant...-3188559/v1.  Neurogenetic alchemy: CAP-Mac unleashes the secrets of primate brains with brilliance ...