...retinal cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved...would express Cre-mCherry and GFP.   Figure 2: Synthetic promoters can be paired for targeted expression...1345-1356. https://doi.org/10.1038/s41593-019-0431-2  Neff, E.P. An AAV library for retinal cell expression...the same transgene: an optogenetic reporter, channelrhodopsin, fused to a fluorescent GFP marker (CatCH-...

...super-resolution or single-molecule techniques.   Figure 2: Two-color super-resolution imaging of ER and mitochondria...stimulated emission depletion (STED) nanoscopy. Scale bar, 2 μm. Inset shows comparison with standard confocal ...enables applications like multi-color imaging (Figure 2). If you’re working with purified proteins in vitro...Labeling in Living Cells. Chemistry & Biology, 15(2), 128–136. https://doi.org/10.1016/j.chembiol.2008.01.007...conjugates: A flexible chemical tag. Nature Methods, 2(4), Article 4. https://doi.org/10.1038/nmeth749 Strebinger...efficiently with SNAP-tag. If you want to use far-red rhodamine dyes for super-resolution imaging, HaloTag gives...

.... The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most people choose...overnight at 4 °C and their secondary antibody for 1–2 hrs at room temperature, which allows them to reduce...Tris-glycine 6–400 kDa Tris-glycine 100 V, 1–2 hours Easy and cheap to handcast Short shelf-life...–40 kDa Tris-tricine 30 V, 1 hour or 100 V, 1–2 hours Good separation, quality, and stability ...separation of proteins of different sizes.  Table 2: Recommended gel percentages for various protein sizes...are consistent and stable at room temperature for 1–2 years and are available in formulas specific to your...response over a broad dynamic range that typically spans 2–5 orders of magnitude, which includes a wide dynamic...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...story (1). From a deep sea shrimp to a compact powerhouse of bioluminescence, NLuc is 100-fold brighter...

... Figure 2: Ace-mScarlet is a fusion between the voltage-sensitive inhibitory rhodopsin Ace2N and mScarlet...Ace-mScarlet, a fusion of a voltage-sensitive inhibitory rhodopsin (Ace2N) with the red fluorescent protein (mScarlet...green-fluorescent GEVIs and blue-light-sensitive channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids...

...by delivering an AAV construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive cation channel... Plasmid tools for microbiome studies Channel rhodopsins for optogenetics Viral Vectors 101: Chemogenetics...

...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...Cas9’. The Cas9 nuclease is a genome engineering powerhouse, and for good reason. It is guided by an RNA ...

...also successfully regulated expression of channelrhodopsin-2, commonly used in optogenetics, opening up... Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation...

...retroviral library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express ...Gottschalk lab has surveyed a range of microbial rhodopsin GEVIs in excitable cells of the nematode Caenorhabditis...membranes, which might beneficially influence the rhodopsin properties. Find these voltage biosensors at Addgene...

... ligands can elicit varying degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic...fluorescent labels. In Nature Methods (Vol. 19, Issue 2, pp. 149–158). Nature Research. https://doi.org/10.1038...and mitophagy, can be tracked using dyes like Rhodamine 123 (R123) (Liu et al., 2017). In multi-color ...diffraction limit of light. Dyes like silicon-rhodamine or Janelia Fluor (JF) dyes have specific properties...

...immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101/gad.2.6.718. PMID: 2843425...Targeted Gene Expression in Drosophila.” Genetics 186(2) (2010): 735-55. PubMed PMID: 20697123. PubMed Central... in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197-214. PubMed PMID: 22129153. Additional...//doi.org/10.1186/1472-6750-6-43Kaczmarczyk A, Vorholt JA, Francez-Charlot A. Cumate-inducible gene expression...

...light sensing proteins. Furthermore, during the first 2 weeks of life, larvae already exhibit distinct behaviors..."Optical control of zebrafish behavior with halorhodopsin." Proceedings of the National Academy of Sciences...

...articles, such as: Sander JD & Joung JK, Nature Biotech, 2 March 2014. Or browse the articles related to the ...be inserted into the genome using CRISPR/Cas forHomologous Recombination (HR)? How long should the homology...

...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...your bioinformatics skills (or your friendly neighborhood bioinformatician) to look at RNA expression ...

...inside the cell,” says Pedre Pérez. Figure 2: Micrographs of the biosensor expressed in the human...method allowed Tian to optimize a red‑shifted archaerhodopsin‑derived genetically encoded voltage indicator...

...dimerization-dependent protein interactions. FEBS letters, 591 2, 415-424. PubMed PMID: 28032891. This is a different...and BioID, any protein that’s within a ~10nm neighborhood of the bait protein will be biotinylated. This...

...products have distinct localizations. Figure 2: Split-wrmScarlet labeling proteins with different...a Cre-dependent, soma-targeting inhibitory channelrhodopsin. It’s now available as AAV5. Find the AAV...

...work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes...Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work/life balance. Is it truly possible to...