...! References Kuhlman, B., Dantas, G., Ireton, G. C., Varani, G., Stoddard, B. L., & Baker, D. (2003)....John Jumper! The Baker Lab focuses on designing entirely new proteins, improving our understanding of how...

...predictable kinetics over 14 hours of expression in C. elegans embryos. However, this initial timer was ...timer could be used in vitro, in mammalian cells, in C. elegans, and in Xenopus embryos. Of course, timers...of protein concentration. Calibration curves correlating this ratio to the total expression time can therefore...used to: Monitor dynamics of vesicle fusion and release from the plasma membrane by fusing various vesicle... situation where one wants to understand the relationship between the age of a cell, protein, or cellular...nascent tissues and the formation of limbs are correlated with changes in gene expression. In a developing...

... a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical abbreviations: Gr= Gracile tract...Deisseroth. (2014) Nat Protoc., 9, 1682-1697. Pubmed. C. Soderblom, D.-H. Lee, A. Dawood, M. Carballosa, et...major advances in light microscopy, particularly related to the neurosciences.  These developments in microscopy...microscopy) (16).  Both 3DISCO and CLARITY have relied on the use of transgenic mice in which a subpopulation... our understanding of the structure-function relationship of the central nervous system under normal and...

...construct. Reactions are often cycled between 37°C and 16°C to provide optimal reaction temps for both the...practices for assemblies of more than 10 modules often rely on two-step hierarchical approaches using different...pre-determined manner. If the original insert religates into its original vector, it will subsequently...

...If the phenotype is corrected, students can be relatively confident that the gene from the test species...procedure adds both V5 and His6 epitope tags to the C-termini of Met proteins, which can be used to confirm...

...calculate the viral titer: GC/mL = {[(R*C)(1000/V)]*D} R = Reaction volume C = Copies/uL V = Volume of virus ...polymerase chain reaction Browse our blog posts related to viral vector protocols and tips Read all of ... by qPCR Browse our collection of viral vector related protocols Learn more about the Addgene viral service...

...Most FP fusions for FRET studies are at the N- or C-terminus of your target protein, but even these modifications...can insert your protein of interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition...for observed FRET efficiencies between 80% and 10%. C Cy3NTA and Cy5NTA bind His tags similarly but with...poly-histidine tag often used for protein purification is a reliable FRET acceptor labeling site when used in conjunction...

...into an attL-containing entry vector.   Method C: Restriction cloning of a restriction enzyme fragment...attL-containing entry vector.   Figure 5: Method C to create an entry clone: Restriction cloning the ...their desired DNA sequences. Gateway technology relies on the two reactions described below: The BP Reaction...

... epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest on the C- or N-terminus. ...conformations and interactions. For difficult to release proteins, including those in the nucleus, more ...

...overhangs Cas13a (C2c2) 2x HEPN crRNA RNA 3' A, U, or C (not required by all orthologs) Specific RNA cleavage...spacer sequence and consists of a single A, U, or C base pair. In bacteria, once Cas13a has recognized...Class 2, candidate 1, 2, or 3). C2c1 and C2c3 are related to Cpf1, but they require both a tracrRNA and crRNA...

...8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune response against AdV. Importantly...4. Mittereder, Nanette, Keith L. March, and Bruce C. Trapnell. "Evaluation of the concentration and bioactivity...

...agarose gel to confirm product presence, size, and relative quantity. End point PCR is most commonly used ...     References and Resources References Shen C-H (2019) Amplification of Nucleic Acids. In: Diagnostic...reagents and several heating and cooling steps. PCR relies on heat resistant DNA polymerase from the thermophilic...

...Nicotiana benthamiana. N. benthamiana is a tobacco relative and a good model for plant disease because it ...assembly of multigene constructs. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. PLoS One. 2011...tools work in any plant? Patron: The MoClo kit is entirely for dicots, not grasses and wheat, but we do work... have your parts. That was the motivation for releasing a large parts kit. Once you have so many parts...

...References and resources Milling, L. E., Markson, S. C., Tjokrosurjo, Q., Derosia, N. M., Streeter, I. S....Cancer and the Immune System: Deciphering the Relationship...

...specificity of GFP expression for the indicated ssAAV5s. C) Quantifications showing high microglia transduction... microglia-targeting AAV plasmids here! Serrano, C., et al. (2023). Simple and Highly Specific Targeting...fast-spiking neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also led to similar...dCas9-VPR, prime editing, and delivery of disease-relevant large proteins. They demonstrated REVeRT in various...

...these methods a try.  References:  Branon, Tess C., et al. "Efficient proximity labeling in living cells...1837. PubMed PMID: 30676735. Kim, D. I., Jensen, S. C., Noble, K. A., KC, B., Roux, K. H., Motamedchaboki...array of biological contexts.  Proximity labelling relies on the labelling of neighboring proteins within...interactions in vivo (Ge et al., 2019). This technique relies on an S. auerus transpeptidase sortase A variant...

...aggregation. Saha et al. Nat Commun. 2023 Feb 2. Study the C terminal domain of TDP-43 to better understand its... atlas for whole-brain neuronal identification in C. elegans. Yemini et al. Cell. 2021 Jan 7. Use a FRET-based...common disorders followed by a list of some of the relevant collections available at Addgene. Amyotrophic ...growing collection of general and neuroscience -related antibodies including: Anti-KChIP1 K+ channel [K55...diagnostics and treatments for Alzheimer’s disease and related disorders. The site curates information on useful...

...IgG2a 177559 Anti-c-Fos [N486/32R] c-Fos Human Mouse IgG2a 177560 Anti-c-Fos [N486/76R] c-Fos Human Mouse...IgG2a 177520 Anti-LRRK2/Dardarin, C-terminus [N241A/34R] LRRK2/Dardarin, C-terminus Human Mouse IgG2a 177521...glutamate receptor Mouse Mouse IgG2a 206619 Anti-c-Fos [N486/25R] c-Fos Human Mouse IgG2a 206620 Anti-Kv1.4 K...220411 LRRK2/Dardarin kinase, C-terminus [N241A/58R] LRRK2/Dardarin kinase, C-terminus Human Mouse IgG2a ...VAPA [N479/111R] VAPA Rat Mouse IgG2a 220414 c-Fos [N486/93R] c-Fos Human Mouse IgG2a 220415 Cavalpha2delta2...] N483/126 Rufy3 Mouse Mouse 182090 C-Fos scFv [N486/32] N486/32 C-Fos Human Mouse 182091 Mortalin/GRP75...84 scFv Rufy3 Mouse Mouse 190560 c-Fos scFv [N486/76] N486/76 scFv c-Fos Human Mouse 190561 BKbeta2 K+...

...groups making use of the new optogenetic tools in C. elegans and in Plasmodium. For those who do want ...those optogenetic controls, gene expression levels reliably increased in vitro by a factor of 10 to 20. A ...modifications in the genome,” Zhang said in MIT’s press release. In an accompanying Nature News & Views Andreas...

...similar nuclease folds found in other proteins. The C-terminal domain (CTD) of the protein also belongs ...https://doi.org/10.1038/s41586-022-04470-1 Cofsky, J. C., Soczek, K. M., Knott, G. J., Nogales, E., & Doudna...Sternberg, S. H., Redding, S., Jinek, M., Greene, E. C., & Doudna, J. A. (2014). DNA interrogation by the...the enzyme needs cellular recycling factors to release its handiwork (Sternberg et al., 2014). New discoveries...