...sequences (either MS2, PP7, or boxB) that each recruits a different fluorescent protein (BFP, GFP, or RFP) to... sgRNA containing alternating MS2 and PP7 sequences (sgRNA-Sirius-4X(MS2-PP7)) that result in yellow fluorescence...recruit GFP to the loci of interest. To determine how the sgRNA-Sirius-8XPP7 and sgRNA-Sirius-8XMS2 scaffolds...) Diagram of sgRNA-Sirius-8XMS2, sgRNA-Sirius-8XPP7, and sgRNA-Sirius-4X(MS2-PP7) showing recruitment ...unstable.   Figure 1: CRISPRainbow labeling. Two MS2, PP7, or boxB sequences inserted into the sgRNA generates...green, or red colors for labeling. A combination of MS2, PP7, or boxB sequences in the same sgRNA will generate...’ end of the gRNA. Because of the success of the MS2 and PP7 RNA hairpin sequences in CRISPRainbow, they...

...Blue MS2-MS2 Green PP7-PP7 Red boxB-boxB Cyan MS2-PP7 Yellow PP7-boxB Magenta boxB-MS2 White...signal to noise ratio of the system, they modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly...the amount of background fluorescence from unbound GFP-dCas9. With these improvements, labeling efficiency...White boxB-MS2-PP7 Despite the success of Ma et al.’s approach, fluorescently labeled Cas9’s come with...sequences, which recruit fluorescent proteins BFP, GFP, and RFP fused to the respective hairpin binding ...Fluorescent Proteins Control Protein Activity with GFP Resources on Asddgene.org Find Fluorescent Protein...

...protein (NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter mRNA via an MS2 coat protein...switching, TRICK requires the following components: a GFP protein that binds to the coding region of a reporter...protein (NLS-MS2-RFP) the reporter mRNA, which contains the tethering sites for the fluorescent proteins and...Plasmid 64542 or 64543) Before translation, both GFP and RFP are bound to the reporter, causing it to ...appear yellow. The act of translation pushes NLS-PCP-GFP off of the mRNA, leaving only a red FP bound to the... contains a nuclear localization signal, NLS-PCP-GFP then returns to the nucleus to bind cognate untranslated...achieved by using multiple copies of the PP7 and MS2 coat protein binding sites to tether many copies ...

...S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness... There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made...examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP can exist in a protonated and...its acid tolerance is maintained in cells.   GFP is big GFP is a 28 kDa protein that resembles a cylinder...2013; Hilaire et al., 2017). GFP can only be used for tagging proteins GFP fusion proteins are made by ... a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps... al., 2018). GFP is not tolerated in my fusion It is standard practice to attach the GFP to the N- or ...

...protein (NLS-PCP-GFP) a RFP protein that binds to the 3’ UTR of the reporter mRNA via an MS2 coat protein ...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...protein Knock-off, is made up of three components: a GFP protein that binds to the coding region of a reporter...protein (NLS-MS2-RFP) the reporter mRNA, which contains the tethering sites for the fluorescent proteins and... coat protein binding sites and 24 copies of the MS2 bacteriophage coat protein binding sites to tether...

...of these probes, fluorophore binding aptamers and MS2-binding fluorescent proteins were the state of the...as compared to imaging with broccoli aptamers or MS2 tags using the same number of fluorophores. You can...real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry fluorescence...involves tracking the ratio of mCherry-positive to eGFP positive cells through fluorescence microscopy or...

...transcription, but with two additional activation domains (MS2 and p65) to enhance transcriptional activation. Zhang...activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 ...

...to recruit an sgRNA with MS2 hairpin binding sites, which then recruit an MS2-fused hyperactive variant..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...with unique tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many ... of DsRed connected to a pH-sensitive variant of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The...

...mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD to decrease methylation and activate transcription...) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR for targeted cytosine methylation...methylation in mammalian cells. Co-expression markers EGFP and PuroR enable sorting and selection of transduced...Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase...epigenome-editing tools. pLV hUbC-dCas9-MQ1(Q147L)-EGFP is also available for lentiviral transduction. Lysine-specific...

...binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag scaffold...scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based...Fragments. IJMS 21:6324 . https://doi.org/10.3390/ijms21176324  Additional resources on the Addgene blog: ...

...highlighted back in 2015, where sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs ...Int J Mol Sci 24:5994. https://doi.org/10.3390/ijms24065994 Xu J, Xu K, Jung S, et al (2021) Nanobodies ...