...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...

...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 ...

...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...

...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 ...

...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...

...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...