...the promoter region of gamma hemoglobin (HGB1/2) leads to continuous expression of fetal hemoglobin even...efficiency, respectively) and outperformed ABE7.10 by at least 1.4 fold. The ability of ABE8.20-m to edit primary...editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464–471 . https://doi.org/10.1038/...Addgene blog Find more blog posts about base editing Learn about another evolved base editor: ABE8e Read about...

...target DNA without cleavage until it's activated by light. Once activated, Cas9 cleavage can induce DSBs ...probes and editors. A probe array may consist of at least two sensor modules targeting a specific chromatin...complex binds to the degron sequence ultimately leading to the degradation of the AID-tagged protein. To...

...spectrum. The experimental apparatus should be cleaned thoroughly between animals as olfactory stimuli...animal (ex: urine) can be distracting. If it is not cleaned, animals tested towards the end of an experiment...coordination to more complex situations such as learning and memory or emotional states, for example anxiety...Guide to Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...Arabidopsis in Education Learning Synthetic Biology with BioBuilder 3D Models for learning about CRISPR Resources... subsequently destroyed by Argonaute-catalyzed nuclease activity or their translation disrupted through...protocols and want to share pictures, feedback etc. please write to [email protected] and if you make...

...Addgene Blog Learn about Synthetic Biology with BioBuilder Use Addgene Videos to Learn about Basic Lab...genes in other species, but these studies generally leave questions about gene function wide open. What if...Check Out Our Molecular Biology Reference Pages Learn Simple Lab Techniques in Our Protocol Videos ...

...unfortunate experience of witnessing…nothing. For some unclear reason, cells are not getting transduced. Here,...reducing the culture volume into which the virus is released, a more concentrated virus solution can be obtained...PMC330758. Additional Resources on the Addgene Blog Learn All about Adenovirus Associated Virus New to Virus...Virus? Check out Our Primer for Beginners! Learn All about Lentiviral Vectors Additional Resources on ...

...serotype, as some rAAV serotypes are abundantly released into the culture media, while others remain cell-associated...and non-genome-containing (empty) capsids, thus leading to an overestimate of transducible units in the...and dehydration (Howard & Harvey, 2017). You can learn more about common AAV applications by reading our...Wilson, J. M. (2010). Efficient serotype-dependent release of functional vector into the culture medium during...

...communication. But first, take a deep breath; at least you know you have an answer to that previous nosy... complicated process even more difficult. I’ve learned that some of the technologies that helped us navigate...application process and usually occurs in the few months leading up to applications opening. With virtual options...be a great opportunity to introduce yourself and learn more about your schools of interest. Benefits of...

...desired recombination event will remove NeoR only and leave exon 2 floxed, as seen in line 4 of Figure 4. Since...tools for creating knockout animals or cell lines, please consider sharing them by depositing with Addgene... with CRISPR through Non-Homologous End Joining Learn How the FLEx Switch Can Be Used to Turn Genes On...Mammalian Expression Vectors Find Cre-Lox Plasmids Learn More about CRISPR  ...

...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko...environmental isolates. For example, Peters and colleagues used Mobile-CRISPRi to knockdown individual genes...-CRISPRi. As a proof of principle, Peters and colleagues constructed a 40-member library targeting 20 ...

...four times in a 95% ethanol and dry ice bath to release the virus particles from the cells. Cell lysates...batch-to-batch variability. Check out this post to learn more about different ways to titer AAV. Do you ...you have any AAV production tips or tricks? Please share them in the comments below! Special thanks ...first time CRISPR user Resources on Addgene.org Learn how Addgene produces AAV and lentivirus Check out...

... blog Learn the basics of Golden Gate assembly Check out NEB's work on ligase efficiency Learn about ...assembly, ligation of mismatched pairs of overhangs can lead to incorrect assemblies, so care must be taken to...is a molecular biologist and a Development Group Leader overseeing a team bringing new products to market...

...remember what to say and provide your audience with a clear picture of your work. Practice, practice, practice...with people inside and outside your field. Your colleagues, lab mates, and PI can provide feedback on your...Anticipate questions: When practicing with your colleagues, ask them to brainstorm potential questions you... of a member of the Addgene depositor community Learn about careers at Addgene ...

...forming noncovalent dimers. This property of EGFP can lead to significant artifacts. If you're using green ...reportedly monomeric FPs are not actually monomeric, at least in practical terms. The propensity of a particular...FP dimerization coupled with POI dimerization can lead to polymerization of the fusion protein. A dimeric... members (superfolder, Emerald, CFP, YFP, BFP, Cerulean, Turquoise, Venus, and Citrine) can be monomerized...

...innate immune system; and reconfiguring of the nuclear compartment, gene expression, and protein translation...responses that inactivate viral genomes or otherwise clear the virus from the cell population (Kafri et al....Adenovirus Tales: From the Cell Surface to the Nuclear Pore Complex. PLoS Pathog, 11(6), e1004821. https...stages of infection for efficient cell lysis and release of adenovirus from infected cells. J Virol, 70(...

...numerous discoveries have been made using the Cas9 nuclease in the absence of genomic alterations. These studies...sequence via a targeting gRNA. But dCas9 does not cleave the DNA. Much of the research using dCas9 has focused...research conducted in the Hodaka Fujii Laboratory, please also see our interview with Dr. Fujii. References..., et al. "A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity." Science337.6096...

...finding how-to videos on YouTube long before he learned (somewhat successfully) how to text or use Facebook... communication that you enjoy and that plays at least slightly to your strengths. Otherwise, you’re going...cost or freeware alternatives and how steep of a learning curve it will be to get to the necessary level...balance out cons. Flexible. Can allow people to learn in multiple ways from the same experience.  Can ...

...research interests include proteomics, machine learning, and visualization for big biology. GPS for the...are critical? One hypothesis: the enhancers! This leads us to a three-step approach for designing an experiment...database. Aneesh's research interests span machine learning, proteomics, and user interfaces for math.     ...Catch Up on Your CRISPR Background with CRISPR 101 Learn about CRISPR Delivery Methods Validate Your Genome...

...and controls (see Tips below), and by validating leads with other methods. More common are false negatives...levels of bait and prey proteins - overexpression can lead to false interactions. Lowered expression levels...techniques! Minimizing false negatives: Interactions leading to detectable reporter signals depend on a number...problems with one or more of these parameters can lead to a high number of false negatives in Y2H screens...

...a cooler. Chances are one of your colleagues has a frisbee at least. Use it! 8. Organize If you’re doing...make an effort to build relationships with their colleagues. It can be easy to get caught up in the minutia...true. If your event requires a lot of set up or cleanup, ask for volunteers to help out. Delegate- it will...