...a FRET acceptor to mRuby2 or mScarlet. Image: Murakoshi et al., 2019. ShadowR is an orange light-absorbing...light-absorbing chromoprotein that Hideji Murakoshi’s lab developed as an acceptor for fluorescence lifetime...https://doi.org/10.1074/jbc.m606921200 Dove SG, Takabayashi M, Hoegh-Guldberg O (1995) Isolation and Partial...1). https://doi.org/10.1186/s13036-018-0100-0 Murakoshi, H., Horiuchi, H., Kosugi, T., Onda, M., Sato,...

...properties once exclusive to small-molecules: red-shifted spectra, ion sensitivity, photoactivation, etc....sample is wholly dependent on the photon budget and pushing the frontiers of fluorescence microscopy often ...can feel like a college student searching couch cushions for spare change, desperate to extract a few more...which is particularly important for samples where washing is difficult, such as intact tissue. Some chemical...

...two independent groups (Komor et al., 2016 and Nishida et al., 2016) invented CRISPR-base editors by tethering...27096365. PubMed Central PMCID: PMC4873371.  2. Nishida, Keiji, et al. "Targeted nucleotide editing using...25733849. PubMed Central PMCID: PMC4371917. 5. Shimatani, Zenpei, et al. "Targeted base editing in rice...PubMed Central PMCID: PMC5972399. 12. Molla, K., Shih, J. & Yang, Y. Multiplex CRISPR-mediated base editing...

...insertion that disrupts fluorescence due to a frameshift. Restoration of fluorescence can only occur through...Harris Lab relies on the restoration of a frame-shift in mCherry that restores fluorescence to monitor...tyrosine generating a GFP variant (GFPY66) that has a shifted emission spectra. Thus a successful base editing...with a cytidine deaminase base editor results in shift in emission spectra from BFP to GFP. Image from ...

...initiation heavily dependent upon the core of a Shine-Delgarno sequence found at the 3’ end of the leader...was predictable based solely on the promoters and Shine-Delgarno sequences used in this so-called bicistronic...Academy of Sciences 113.9 (2016): 2388-2393. Huang, Shijie, Xuechen Zhu, and Charles E. Melançon III. "Detection...ChemBioChem 17.10 (2016): 895-899. Choudhury, Rajarshi, et al. "Engineering RNA endonucleases with customized...

...for. Secondly, categorize your edit – is it any frameshift mutation? A specific insertion or deletion? A...line is desired, any mutation that introduces a frameshift in the coding region of interest will get the...and run the PCR product on a gel to look for size shifts. Once you’ve identified homozygous clones, however...large so that you can easily visualize the size shift you anticipate. A good rule of thumb is to keep ...

...own piece (3). After incubation, blocking, and washing, the membrane is reassembled and imaged. In this...(PBS) are the most commonly used solutions for washing. If your background noise is high, you can consider...Like blocking, you don’t want to skimp on your washing steps! It’s important to let them wash for the ...sequence not accurate Check plasmid sequence for frameshifts and stop codons Weird background patterns...primary antibody at 4 °C overnight Incomplete washing Ensuring wash is happening under sufficient agitation...

...blue-shifted fluorescent protein (donor) and measuring the resultant emission from the red-shifted fluorescent...imaging. The solution is to couple Nluc with a red-shifted fluor. Here, NLuc internally excites the fluor ...these tools, NanoLuc-fluorescent protein fusion shifts NLuc’s bright blue luminescence to wavelenghts ...

...about fighting scientific partisanship, including more scientists in our leadership, and raising awareness...McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072°N 77.0369°W – Mark Lucera I...science education. Photo cred Alina Chan. Seattle, Washington, USA 47.6062°N 122.3321°W– Justin Sherman Prior...

...these plasmids for the majority of the countries we ship to, first on a temporary and now on a more lasting... exceptions for rabies and VSV-G genes, plasmids ship out approximately three weeks faster, as no license...addgene.org for the most-up-to-date information on shipping plasmids to your country. Curious about how rabies...

...monitor protein and cell ageing, as the spectrum shift occurs during Timer protein life due to a form of...read the Addgene interview with Mark Howarth. Shining a light on channelrhodopsins – Chronos & Chrimson...spectral peak at 590 nm, which is 45 nm more red-shifted compared to other variants. The group further optimized...functional GFP, and the cells will turn green. Mashiko et al., Sci Rep 2013 Nov 27; 3:3355. Rinehart...TALEN assembly and activity? The laboratory of Takashi Yamamoto has created a complete TALEN assembly ...

...and watch it now! (Bonus: Tim kindly offered to finishing answering all submitted questions via blog post... the next few weeks.)  Finally, Addgenies love sunshine and warm weather - check out our Instagram all...endeavors - in between some much-deserved vacations, sunshine, and beach trips, of course.    *not me, though...

... FACS machine after a single use. So how about washing and reusing pipette tips in a high-throughput and...example, offers automated, benchtop, high throughput washing devices that are compatible with a variety of pipette...incentives (for example, by grant agencies), we can shift toward a more sustainable one. The math on this ...

...of the method. One observation was an inverse relationship between deletion size and frequency [3]. There...coding sequences often result in loss-of-function frameshift mutations. However these may require restriction...genomic landscape may be surveyed in unprecedented fashion. Targeted deletions are just one of many available...

...repository in 2023.  In June, we announced a new partnership with the Institute for Protein Innovation (IPI...we’re pleased to announce that we are now able to ship rabies plasmids in our repository to international...through active conversation and collaboration.  Wishing you a healthy, productive, and thriving 2024, mirroring...

...muscular atrophy patients. Noushin Nabavi - Peritoneal Mesothelioma  Noushin Nabavi is a Postdoctoral Fellow...diagnostic tools and animals models of the disease in partnership with Taconic Biosciences. The study of peritoneal...

...Expanding your network of relationships early and often is the most effective tactic a scientist in training...result of existing professional (and personal) relationships. However, most job seekers will and should apply...after the academic bench. Utilizing existing relationships as part of the job application process can dramatically...

...person, and helps improve your interpersonal relationships with people you’re collaborating with remotely...like something out of your nightmares, shared ownership of materials produced from your collaboration ...the project. The state of research is constantly shifting, and long-distance collaborations are becoming...

...awesome. The process of writing, editing, and publishing blog posts runs its course more quickly than ...supervisors warned me, the swift gratification of publishing posts can be addicting. Even more satisfying,...always thoroughly enjoyed: travel. I’m actually finishing up this post on a plane back to Boston from Kentucky...

...deaminase from sea lamprey fused to Cas9 nickase (Nishida et al., 2016). Target-AID acts similarly but not...efficiency but higher indel formation than BE2 Nishida et al., 2016 Target-AID   Edits 3-5 base window...685. https://doi.org/10.1038/s41587-022-01533-6 Nishida, K., Arazoe, T., Yachie, N., Banno, S., Kakimoto...Mochizuki, M., Miyabe, A., Araki, M., Hara, K. Y., Shimatani, Z., & Kondo, A. (2016). Targeted nucleotide editing...