...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 ...

...been fused to the MS2 binding sites within this plasmid. pZE31-DsRed-ms2x2 26248 MS2 coat protein binding...RNA-binding protein, MS2. Can be used in combination with a fluorophore-tagged MS2 (such as that found ...found in pZS*12-MS2-GFP ) to track RNA localization of your gene of interest once its appropriate cDNA has...RNA-binding protein MS2 fused to dsRED. Can be used in combination with a fluorophore-tagged MS2 (such as that...that found in pZS*12-MS2-GFP ) to track RNA localization of your gene of interest once its appropriate ...many different restriction enzymes. pPro24-gfp contains GFP but all others are empty. pdCas9-bacteria ...can be used in super resolution microscopy. pZE31ms2 25860 MS2 coat protein binding sites Philippe Cluzel...

...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 ...

...Zhang 61426 lenti MS2-P65-HSF1_Hygro MS2-P65-HSF1 Hygromycin lenti vector encoding the MS2-P65-HSF1 activator...Purpose PI 17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 ... complex with a 2A Hygro resistance marker (EF1a-MS2-p65-HSF1-2A-Hygro-WPRE) Zhang Pooled Barcoding Libraries...analysis of clonal dynamics. This library expresses EGFP for easy visualization via direct fluorescence. ...

...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...

...activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with 2A GFP 61424 : sgRNA...sgRNA incorporating two MS2 RNA aptamers at the tetraloop and stemloop 2 The MS2-P65-HSF1 activation helper...(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH fusion ...sgRNA cloning backbone with MS2 loops at tetraloop and stemloop - Contains BbsI sites for insertion of guide...resistance marker 61426 : lentiviral vector encoding the MS2-P65-HSF1 activator helper complex with a 2A Hygro...marker 61427 : lentiviral sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance...PX552; U6 promoter-driven; for sgRNA cloning; has GFP-KASH for FACS sorting SaCas9 + single guide RNA: ...

...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...

... pCAG-SpCas9-GFP-U6-gRNA 79144 Mammalian hU6 yes, cut S. pyogenes EGFP Zou pCAG-eCas9-GFP-U6-gRNA 79145...sgRNA(MS2)_zeo backbone 61427 Mammalian/Lentiviral BsmBI none S. pyogenes Zhang pL-CRISPR.EFS.GFP 57818...Cerulean Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA...cut Lb Cpf1 Neo Welker AIO-GFP 74119 Mammalian U6x2 yes, nick S. pyogenes EGFP Jackson AIO-mCherry 74120...Mammalian BbsI yes, cut S. pyogenes Puro Zhang sgRNA(MS2) cloning backbone 61424 Mammalian BbsI none S. pyogenes...Lentiviral hU6 none S. pyogenes Puro Moffat lenti sgRNA(MS2)_puro backbone 73795 Mammalian/Lentiviral U6 none...yes, interfere S. pyogenes URA3 Davis lenti sgRNA(MS2)_puro optimized backbone 73797 Mammalian/Lentiviral...

...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...

... PINK1 N-GFP PINK1 GFP CMV Parkinson's Mark Cookson 13316 pcDNA-DEST47 PINK1 C-GFP PINK1 GFP CMV Parkinson's...21190 GFP-pcDNA3-PKCgamma-cys1Acys1B PRKCG GFP CMV Spinocerebellar ataxia 27 Tobias Meyer 21204 GFP-N2-PKCgamma...PKCgamma PRKCG GFP CMV Spinocerebellar ataxia 26 Tobias Meyer 21205 GFP-C1-PKCgamma-C1A PRKCG GFP CMV Spinocerebellar...-HtrA2-EGFP HTRA2 GFP CMV Parkinson's L. Miguel Martins 14122 pcDNA3-HtrA2-EGFP S306A HTRA2 GFP CMV Parkinson's... GPD 25QDProGFP p416 HTT GFP GPD Huntington's Susan Lindquist 15569 GPD 104QDProGFP p416 HTT GFP GPD Huntington's...GAL 25Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15581 GAL 46Q+ProGFPp416 HTT GFP GAL1 Huntington's...GAL 72Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15583 GAL 103Q+ProGFPp416 HTT GFP GAL1 Huntington's...

...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 ...

...48201 CAG-GFP-IRES-CRE Cre and GFP coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG...116879 CAG-Cremyc-2A-GFP GFP and Cre CAG Mammalian Lu 117148 Hiv7CMV-Cremyc-2A-GFP GFP and Cre CMV Lentiviral...iRFP670-EFS:Cre-2A-GFP iRFP670, Cre, and GFP TRE Mammalian Jacks 68544 AAV-Cre-GFP Cre CMV AAV Nestler...13770 pCALNL-GFP Cre dependent GFP expression Mammalian Cepko 8389 p212 pCMV-EGFP/RFP EGFP-dsRed gene switch...promoterless CRE-GFP fusion none Mammalian Sauer 11960 pBS537 tet-hCMV-GFPcre tet inducible Cre-GFP fusion tet-hCMV... CMV Lentiviral Fuchs 26646 pCAG-Cre-IRES2-GFP Cre and GFP coexpression CAG Mammalian Chenn 26647 pCAG-Cre...CAG Retroviral Gage 49056 AAV-GFP/Cre Cre-GFP fusion CMV AAV Gage 49111 pEMS1980 iCre with MCS for inserting...

...fluorescent marker like green fluorescent protein (GFP), creating a customizable DNA or RNA label for fluorescence...fluorescence microscopy. RNA aptamers, like MS2 or PP7, can also be fused to the gRNA to recruit fluorescently-tagged...

...fluorescent marker like green fluorescent protein (GFP), creating a customizable DNA or RNA label for fluorescence...fluorescence microscopy. RNA aptamers, like MS2 or PP7, can also be fused to the gRNA to recruit fluorescently-tagged...