...costly to share. What is a RANbody? A RANbody is a fusion between a reporter and a nanobody that’s encoded...J.R., & Yamagata, M. (2018). Reporter-nanobody fusions (RANbodies) as versatile, small, sensitive immunohistochemical...

.... Historically, glutathione-S-transferase (GST) fusion proteins have been immobilized on a reduced glutathione...glutathione matrix. The advantage of a fusion protein is the efficient purification of an already immobilized...for kinetic analysis Proteolytic removal of the fusion protein could provide a more relevant kinetic analysis...produces biotin ligase and a protein of interest fused to maltose binding protein (MBP, New England BioLabs...protein in bacteria and subsequently purify the fusion protein from a bacterial extract. The MBP tag can...using Xa protease. The carboxyl-terminus of the fusion protein includes an AviTag and a 6X HisTag for ...protein we tested, it may be possible with other fusion proteins - but it would remove both the AviTag ...

...Fluorescent Protein Fusions Observing a bright signal for the first time after cloning an FP fusion with your favorite...consequences of misfolded dark FP fusions including erroneous quantitation of fusion protein levels. It remains...misfolding impacts fusion protein function and this could lead to under and overestimation of fusion protein activity...impact of dark, non-fluorescent, and misfolded FP-fusions on quantitative imaging experiments and cell physiology...interest is exciting and rewarding. But, what if an FP fusion with a resident endoplasmic reticulum (ER) protein...troubling, what if a significant fraction of the fusion protein molecules fail to correctly fold? Misfolded...localization and dynamics of fluorescent protein fusions in live cells in real time. Previously, we have...

...engineering of fusion proteins is relatively straightforward. However, generating a fusion while keeping...Creating fusions at the N- and C- termini of your protein of interest The most straightforward fusion protein...use terminology like “C-terminal fusion,” but I find this very confusing because it can mean that the fluorescent...cloning site for generating fusions. For the POI-fluorescent protein fusions, our experience is that a ...For the fluorescent protein-POI fusions, we generate straight fusions, without any linker. When both orientations...example, the APT1-mVenus fusion localizes at the Golgi, whereas the mVenus-APT1 fusion does not (figure 2)....fluorescent protein fusions There are several choices to consider before generating a fusion protein, e.g. ...

...Yardena Samuels 29591 426Gal-FUS FUS GAL1 ALS Aaron Gitler 29592 426Gal-FUS-YFP FUS YFP GAL1 ALS Aaron Gitler...Gitler 29593 416Gal-FUS-YFP FUS YFP GAL1 ALS Aaron Gitler 29594 416GPD-FUS-YFP FUS YFP GPD ALS Aaron Gitler...416Gal-SV40NLS-FUS-YFP FUS YFP, SV40 NLS GAL1 ALS Aaron Gitler 29596 426Gal-FUS-1-170aa-YFP FUS YFP GAL1 ALS...29597 426Gal-FUS-1-270aa-YFP FUS YFP GAL1 ALS Aaron Gitler 29598 426Gal-FUS-1-373aa-YFP FUS YFP GAL1 ALS...29599 426Gal-FUS-1-413aa-YFP FUS YFP GAL1 ALS Aaron Gitler 29600 426Gal-FUS-1-453aa-YFP FUS YFP GAL1 ALS...29601 426Gal-FUS-1-501aa-YFP FUS YFP GAL1 ALS Aaron Gitler 29603 426Gal-FUS-101-526aa-YFP FUS YFP GAL1 ALS...29605 426Gal-FUS-267-526aa-YFP FUS YFP GAL1 ALS Aaron Gitler 29606 426Gal-FUS-285-526aa-YFP FUS YFP GAL1 ...

... them to a tumor - they used focused ultrasound (FUS) to heat the tumor area, as the image below shows...stable levels for 6 days - after just 15 minutes of FUS.  Similar constructs to the one above can be made...

...events that occur in less that 5% of the population (Fu et al. 2013, Vouillot et al. 2015). Meanwhile, Sanger...sequence requirement at the terminal 3’ PAM site (Fu et al. 2013; Pattanayak et al. 2013). This approach...magazine.scienceforthepeople.org/.     References 1. Fu, Yanfang, et al. "High-frequency off-target mutagenesis...

... three different sites in the EGFP reporter gene (Fu et al., Nat Biotechnol. 2013). The expression plasmids...vector ( JDS246 ). Additional details can be found in Fu et al., Nat Biotechnol. 2013 and users are encouraged...

... to modify four different endogenous human genes (Fu et al., Nat Biotechnol. 2013). The expression plasmids...vector ( JDS246 ). Additional details can be found in Fu et al., Nat Biotechnol. 2013 and users are encouraged...

...guides (19-20bp)?  Small-scale studies (Tsai et al., Fu et al.) suggest 17-19 bp long guides have reduced...PMID: 25513782. Pubmed Central PMCID: PMC4320685. 3. Fu, Yanfang, et al. "Improving CRISPR-Cas nuclease specificity...

...sites in various endogenous zebrafish genes (Hwang & Fu et al., Nat Biotechnol. 2013). The expression plasmids...pMLM3613 ). Additional details can be found in Hwang & Fu et al., Nat Biotechnol. 2013 and users are encouraged...

...target sites in endogenous zebrafish genes (Hwang & Fu et al., Nat Biotechnol. 2013). The Cas9 expression...expression of Cas9 in zebrafish can be found in Hwang & Fu et al., Nat Biotechnol. 2013. The guide RNA ( gRNA...

...Nup153 Nuclear pores 114410 FUS-mEGFP AICSDP-64 mEGFP RNA-binding protein FUS Paraspeckles/stress granules...

...Consortium (SGC) Plasmids Opher Gileadi Lab plasmids Haian Fu Lab plasmids The TREAT-AD centers are focused on ...

...modifying endogenous genes in zebrafish (Hwang and Fu et al., Nat Biotechnol. 2013). The Joung lab has ...

...PMID: 26829887. PubMed Central PMCID: PMC4735761. 6. Fu, Y. et al. High-frequency off-target mutagenesis ...

...Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F. Cell...

...10.1016/j.neuron.2010.11.021  Gambarotto D, Zwettler FU, Guennec ML, et al (2019) Imaging cellular ultrastructures...

...him on Twitter.   References Arribere JA, Bell RT, Fu BXH, Artiles KL, Hartman PS, Fire AZ (2014) Efficient...

...one of the following genes: SOD1, C9ORF72, TARDBP, FUS and OPTN. Huntington's Disease Huntington’s disease...expression of diphtheria toxin receptor (DTR)–GFP fusion protein. Azim et al. Nature. 2014 Apr 17. Next-...