...want a low affinity calcium biosensor, so that the (1) binding of calcium, (2) allosteric rearrangements... to a red fluorescent state only by simultaneous (1) user-controlled illumination with violet light and...fluorescent indicators. Nature protocols. 2006 Aug;1(3):1057-65. Resources on Addgene.org Addgene’s AAV...GCaMP6 only). The genetic elements required for Drosophila studies have been added for jGCaMP7 and jGCaMP8...

...and drawbacks of four split AAV vector approaches. 1. Overlapping  Overview: The overlapping strategy has...Adeno-Associated Virus Capsids. Human Gene Therapy Methods 27:1–12 . https://doi.org/10.1089/hgtb.2015.140 Frischmeyer...deliver the mini-dystrophin gene to a mouse model of muscular dystrophy. While mini-dystrophin expression levels...levels were lower than expression of wild-type dystrophin in normal muscles, mice treated with the overlapping... trans-splicing vector to deliver a 6 kb mini-dystrophin gene to a mouse model of muscular dystrophy. ...achieved ~80% of muscle cells expressing mini-dystrophin.  3. Hybrid  Overview: The hybrid approach combines...Gene Therapy of mdx Mice With Large Truncated Dystrophins Generated by Recombination Using rAAV6. Molecular...

...Zhang et al., 2015). In this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin...splitting cGAL4 in two and binding each half to a gp41-1-N-intein. Split inteins are protein domains that naturally...mutations with unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust...by members of the worm community but there are Drosophila and zebrafish articles published there as well...

...reconstitution with a normally functioning T cell repertoire (1). In this example, it was possible to transduce the...significant improvements in vision over a period of 1 to 3 years – the treatment appears to be both safe...research and science communication.     References 1. Gaspar, H. B., S. Howe, and A. J. Thrasher. "Gene...directed at diseases including but not limited to hemophilia B, LPL deficiency, Cystic fibrosis, Muscular ...

...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference... plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan, A., ... used arg1 to in vivo image tumour-homing S. typhimurium strain ELH1301. Bourdaeu et al see this as just...

...the other (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv. ...intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged with 10-24 copies...Application. Clinical and Developmental Immunology 2012:1–15 . https://doi.org/10.1155/2012/980250  Bates A,...without disulfide bonds, made by molecular evolution 1 1Edited by I. A. Wilson. Journal of Molecular Biology...made up of glycine and serine with dispersed hydrophilic residues for increased solubility (Monnier et...

...expression inducible by red light. pCph8 50552 pLtetO-1 Anhydrotetracycline Jeffrey Tabor Anhydrotetracycline... pZE31ms2 25860 MS2 coat protein binding sites Philippe Cluzel RNA localization: Contains a 2x array of...DsRed-ms2x2 26248 MS2 coat protein binding sites Philippe Cluzel RNA localization: Contains a 2x array of...Fluorescent protein, GUS, and LacZ Derivatives avaliable Philip Poole A family of promoter probe vectors incorporating...

...biosynthesis in Drosophila melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed...stories. Six spooky science stories Zombie ants Ophiocordyceps unilateralis is a fungus that infects ants ... a set of cytochrome P450 genes discovered in Drosophila melanogaster (Gilbert, 2004). Mutations in these...blood out. Once the blood is depleted, the tongue atrophies and falls off. The louse remains becoming the ...

...This gives each guide a score ranging between 0 and 1. The cutting frequency determination (CFD score) is...converted to a weight (which ranges between 0 and 1) so that they can be used in the rank equation above...converted to a weight (which ranges between 0 and 1) (shown in the graph below) so that it can be used...HDR. Optimal mutation in-depth One the most sophisticated features of CrispyCrunch is how it mutates guide...

...biological questions. The two most common uses are: 1) to test the expression level in a specific system...brightness is compared to that of EGFP which is set as 1. Some proteins are very dim (e.g. TagRFP657, which...Photostability can be as short as 100ms (EBFP) or as long as 1 hour (mAmetrine1.2). However, for most FPs it is a...emission is green or red light. For example, T-Sapphire, LSSmOrange, and LSSmKate. Fluorescent Sensors...

...References 1. Peters, Nathan C., et al. "In vivo imaging reveals an essential role for neutrophils in leishmaniasis...do researchers track fluorescently labeled cells? 1. Epifluorescence microscopy This technique uses a ... The video above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph...infection with S. aureus (red). You can then see neutrophils swarming around S. aureus and eating the bacteria...two-photon microscopy include: How do T cells and/or Neutrophils travel to the sites of infection or damage (Peters...Good two-photon probes when using a standard Ti:Sapphire Laser include: Blue-green fluorescent proteins...this requires other lasers than the standard Ti:Sapphire laser. It is also important to finely tune the...

...organoid model (Sun, et al. 2021).  Figure 1: Example schematic from the Organoid EasyTag system...without altering that of the internal control (Fig 1). Because this method relies on the fluorescence ratio...and can be used for genome editing in moderate thermophiles.  These evolved SpCas9 variants have improved...

...more tools for plasmid mapping and design (Figure 1). Benchling: While you might think of Benchling as...Davis from the University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid...patterns, post-translational modifications and polymorphisms Codon optimization tool: To access this tool...

... that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible dimerization domain...expression needed for most applications.   Figure 1: With blue light, the VVD domains of the BLADE dimerize...Mustafa Khammash's lab at ETH Zürich. “Instead of a graphical abstract, it’s a musical abstract,” says Di Ventura...

.... In literary terms, conflict can be defined as (1) the struggle between two opposing forces or (2) the...journey even when they know the ending.    Fig. 1: We all know that the superheroes win in the end. ...other affected people, man/animal vs nature, anthropomorphized animals, or even seemingly unrelated stories...

...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ...analyzing protein–protein interactions in live Drosophila. eLife 7. https://doi.org/10.7554/elife.38853...

...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion...genes: deletion mapping functional domains of Drosophila hsp 70." The EMBO journal 3.13 (1984): 3087. ...

...another great year of sharing!   References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide...

...sequences were initially discovered in E. coli in 1987 (1), the concept that these clustered repeat sequences...neutralize CRISPR/Cas. Earlier this year, expression of 1 of 5 anti-CRISPR genes were found to inactivate the...light until 2007. Initial experiments exposed S. thermophilus to predatory phages to test if the exogenous...

...has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million bp depending on the polymerase...analyzing the gene’s function. The process is shown graphically in the following figure, in which we add EcoRI...