...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...

...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 ...

...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...

... 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...

...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 ...

...ride is a great activity for both exercise and clearing your mind. And because the outdoors is so huge...events. Download Addgene’s Science Career guide to learn more about career paths and things you can do to...Plan out meals and snacks ahead of time.  Find learning activities for kids. For example, Skype a Scientist...collected into figures for papers or presentations. Learn a new skill for analyzing and visualizing your data...introductions and literature reviews could make the months leading up to your defense less stressful. Think about...

...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...

...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...

...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 ...

...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...

...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...

...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...

... 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...

...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...

...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...

...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...

...it’s become clear that the power of this editing platform lies in the diversity of nucleases and their .... For more mechanistic details on HDR and NHEJ, please see the linked blog posts. Compared to its counterparts...compare to CRISPR sticky-end insertion using the nuclease Cpf1. Since Cpf1 cuts in a staggered pattern, ...Experiment Additional Resources on Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find ...

...Identifying which genes are important for a phenotype can lead to a wealth of additional experiments investigating...consists of two components: the non-specific endonuclease (Cas9) and a single stranded guide RNA (gRNA...to enrichment or depletion of specific gRNAs can lead to skewed results. Selecting a cell type - Theoretically...validation practices, CRISPR libraries can help you learn a lot about your phenotype of interest. For more...Lentiviral CRISPR Libraries” (McDade et al., 2016). To learn a bit about how CRISPR/Cas9 can be used in other...

...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 ...