...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...

...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...

...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 (Link opens in a new window) The TREAT-AD...

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

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

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

...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-...

...://doi.org/10.1016/j.tibs.2023.10.006 Tang, Y., & Fu, Y. (2018). Class 2 CRISPR/Cas: an expanding biotechnology...the different types of CRISPR systems can be as confusing as reading Shakespeare for the first time. The...

...Human Cells. Cell Rep . 13(3):621-33. PMID: 26456817 Fu Y, Sander JD, Reyon D, Cascio VM, Joung JK. 2014....convert adenosine to guanosine. RNA editors : Cas13 fused to the ADAR2 deaminase convert adenosine to inosine...modifications at specific loci. Activate or Repress : dCas9 fused to transcriptional activators or repressors can ...transcription in a reversible manner. Epigenetics : dCas9 fused to epigenetic modifiers can modulate transcriptions...

...Molecular Cell , 73 (4), 714-726.e4. PMID: 30581144 Fu, Y., Foden, J. A., Khayter, C., Maeder, M. L., Reyon...Nature Biotechnology , 31 (9), 822–826. PMID: 23792628 Fu, Y., Sander, J. D., Reyon, D., Cascio, V. M., & Joung...Transcriptase (M-MLV RT) is fused to the C-terminus of Cas9 H840A nickase. The fusion enzyme can catalyze targeted...dCas9-VPR - dCas9 fused to several different activation domains in series; a single fusion protein, instead...constitutive expression of the dCas9 fusion and/or expression of multiple fusions. The effects of CRISPRi/a may...Small Edits Large Edits Genome-Wide Screening Cas9 Fusion Tools Cas9 Alternatives Resources Plan Your Experiment...This means proteins with other functions can be fused to a dCas9 and delivered to specific genomic locations...

...frame with tags or fusion proteins in the recipient plasmid (if you are creating a fusion protein) Ideally...insert into your vector Conduct a DNA Ligation to fuse your insert to your recipient plasmid. We recommend...