... Their reporter system is comprised of two plasmids: Luc (intronless; CMV-LUC2CP/ARE and Luc-I (intron-containing...to the widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have... occur in Luc-I, the produced luciferase is truncated and lacks enzymatic activity. The Luc plasmid (intronless... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs...customized fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single ...fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash and pcDNA3.1+-...

...Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label of cells, there's...Incorporation into a FRET biosensor A FRET biosensor is a reporter that undergoes a conformational change to bring...help you make better fluorescent fusion protein reporters for your studies and greatly lower your chances...be forming dimers in cells. Why should you care? Three simple ways a dimeric FP could ruin your day (and...FP dimerization. Equally problematic, several reportedly monomeric FPs are not actually monomeric, at ...Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...

... Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance... editing. eGFP reporters for single base editing by APOBEC-Cas9 Though the ACE reporter quantifies APOBEC-Cas9...DSB in their previous reporter, the Harris Lab created a panel of eGFP reporters to quantify the on-target...editing reporters. Image from St. Martin et al., 2019. The Harris lab used these eGFP reporters to analyze...Transient reporter for editing enrichment (TREE) The Wang and Brafman lab developed a transient reporter for...cells segregated solely by a reporter of transfection, which only report the efficiency of plasmid delivery... functional reporter system to identify and enrich for base-editing activity The reporters mentioned above...

... and a UAS-driven luciferase reporter construct. In the absence of GFP, no reporter output was observed... and a reporter construct into the mouse retina. GFP expression robustly activated the reporter gene TdTomato...and Cre-DOG, respectively, repurpose popular GFP reporter lines for more sophisticated experimental manipulations...GFP. They then tested T-DDOGs with two mouse GFP reporter lines; again, TdTomato was seen only in cells ...tested in retinal electroporation studies, DsRed reporter expression was induced in ~76% of GFP+ cells, ...Once they’d found suitable pairs, they constructed three components: GBPa-VP16 (activation domain); GBPb-...create a complete transcription factor, activating luciferase transcription. Tang et al. coined this system...

...What are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used... to study gene expression in cells. But these reporters have limited use in vivo because they are based...expression in vivo.  But what ultrasound lacked was reporter genes that could produce ultrasound contrast. ...our lab - the Shapiro Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes that, when...5202–5209 (2017). Bourdeau, R. W. et al. Acoustic reporter genes for noninvasive imaging of microorganisms... Hurt, R. C. et al. Genomically mined acoustic reporter genes for real-time in vivo monitoring of tumors...and 5 g/L NaCl) or on LB-agar plates containing glucose and L-arabinose at 37oC for at least 12 hours, ...

...maintaining fluorescence. The protein structure, first reported in 1996, is an eleven β-sheet-containing “barrel...constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a promoter of...a given cell type. This type of transcription reporting was among the earliest uses of GFP. Fluorescence...micrometastases in nude mice in cancer studies, act as a reporter for DNA double strand break repair, and to label...number of functions, including: tagging genes for elucidating their expression or localization profiles, acting...chromophore formed from a maturation reaction of three specific amino acids at the center of the protein...application Almost as soon as its sequence was elucidated, scientists began engineering new versions of...

... a cleavage free manner. The dCas9-SSAP system reports an efficiency of up to 20%, comparable to commonly... For a one kilobase insertion, the researchers reported an editing error rate of less than 0.3%, both ...using mStrawberry, localization peptides, and reporters that drive the expression of the fluorescent protein...response element (TRE) or kinase translocation reporters (KTRs). Find visual barcode plasmids here! Kaufman...reduced error rate for long sequence knock-ins by: Lucie Wilson Le Cong’s lab has developed a method using... subcellular localization peptide. By combining three different localization peptides with four different...

...that uniquely express selectable and/or sortable reporters in certain states of interest. For example, in... (Sanger sequencing, qPCR, and fluorescent reporter-based) each come with their advantages and disadvantages...the most convenient strategy is the fluorescent reporter based method since it lends itself to cheap, quick...detection with flow cytometry. However, expressing reporters will consume cellular resources and therefore ...constructed and tested eight two-input RSMs and three three-input RSMs. The plasmids used to construct these... green fluorescent proteins in each state. The three methods of state interrogation mentioned above (...PBAD), and the A118 gene downstream of the diacetylphloroglucinol (DAPG)-inducible promoter (PPhlF).  Output...

...Russell and Hirata from the University of Washington reported a surprising finding - when delivered via AAV ...take up ~6.5 kB if delivered in a single AAV. They report roughly 40% repair efficiency in the K562 leukemia...AAV vector cures haemophilia B mice." Scientific Reports 7.1 (2017): 4159. PubMed PMID: 28646206. PubMed... gene cassettes using AAV donor vectors." Cell reports 20.3 (2017): 750-756. PubMed PMID: 28723575. PubMed...underwent homologous recombination at a frequency three orders of magnitude higher than identical targeting...essentially all of an AAV’s ~4.5 kB packaging capacity. Luckily, researchers have devised methods to deliver CRISPR...

... phage promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, ...preventing repressor cI synthesis and allowing reporter gene transcription to take place. Addgene also...Below, you’ll find a short description of these three types of inducible promoters, and examples of each...I2 sites. Supplementing cell growth media with glucose decreases cAMP and represses pBad, decreasing promoter...better option since expression can be repressed by glucose. Temperature inducible promoters are also known...

...even been linked to some diseases. It has been reported that 5-10% of human genes contain a region where...EMBO Rep 1:18–23 . https://doi.org/10.1093/embo-reports/kvd001 Additional resources on the Addgene blog...different codons that encode 20 amino acids and three stop codons, meaning that the same amino acid can...One part of the tRNA contains the anticodon,  a three nucleotide sequence complementary to the mRNA sequence...the 64 codons that encodes the 20 amino acids and three stop codons. (B) Example of codon usage among different...example, Henry Lester’s lab optimized the C. elegans GluCl ion channel genes for expression in mammalian cells...2003) Codon optimization of Caenorhabditis elegans GluCl ion channel genes for mammalian cells dramatically...

...phosphatase-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked mCherry...are available at Addgene. Kim, et al. Stem Cell Reports. 2015. PubMed PMID: 25772473 Targeting and manipulating...enhancer elements and combined it with a variety of reporters (i.e. eGFP) and effectors (i.e. DREADDs). The ...reagents shared via Addgene. Narayan S, et al. Cell Reports. 2017. PubMed PMID: 28813671 Listen to the podcast...fluorescence, brightness, and proper maturation. Three new RFPs were isolated and characterized: mScarlet...lifetime of 1.3 ns, quantum yield of 0.20). All three mScarlet variants have shown great performance in...subtilis libraries Transcription factor fusions elucidate iPSC reprogramming mechanism The Ptashne lab ...

...diseases and metabolic diseases are also underway and reported results are encouraging. Importantly, a significant...combined with AAV for in vivo gene editing, and reports suggest that the first clinical study combining...another genetic disease more prevalent or life-threatening (cystic fibrosis, hemophilia, muscular dystrophy... mid-90's with a lot of work and a little bit of luck A pioneer in the field, Dr. Jean Bennett started...that faithfully recapitulated the human condition. Luck brought all these key elements together at the right... this limitation, with limited success so far. Luckily for patients, this gene can be targeted to RPE ...

.... Andrew and Maria were encouraged by previous reports that described magnetic responses of cryptochrome...reproduced their EGFP findings since the initial report in 2023.)     Figure 1: Fluorescence intensity...a starting point for developing useful tools. A lucky start As Andrew and Maria explained, “to our surprise...directed evolution and screening experiment with three of the proteins that had shown magnetic responses...develops. And though it took a lot of work (and some luck) to get here, he believes MagLOV is just a starting...

...PMM2 gene. Videira’s group recently reviewed and reported that PMM2-CDG patients suffer from immunological... construct NFIX expression vectors as well as reporters of NFIX function. They will then mutagenize the...inherited metabolic diseases with severe and life-threatening consequences. They result from defects in different...these related syndromes. Many thanks and best of luck to the 2018 BeHeard Award winners! Paula Videira...

...have been around since 2000 when Terskikh et al. reported the production of dsRed E5. This timer predictably...fluorescent timers that change color from blue to red report on cellular trafficking." Nature chemical biology... et al’s directed mutagenesis also produced the three separate timers shown in the table: Fast-FT, Medium-FT...lineage-control network that differentiates human IPSCs into glucose-sensitive insulin-secreting beta-like cells." Nature...

...in competition with other similar integrins, as reported by IPI and the Springer lab (Hao et al., 2024)...In this study, we, along with the Springer lab, reported the binding specificity, kinetics, and affinity...environment and the cytoskeleton. Most integrins have three conformational states: bent-closed, extended-closed...processes.   Figure 1: Integrins can shape into three conformations: (A) bent-closed, (B) extended-closed...cells to spread into symmetrical circles. While elucidating the role of tension, however, the researchers...individually and in combination, showing that among the three αV integrins expressed on the BJ-5ta fibroblast ...

...sub-cellular localization signals, plant markers, and reporters (Engler et al., 2014). This kit is a great starter...into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This allows for the multiplexing of sgRNAs...Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media...plant biologists, Addgene has not one, not two, but three CRISPR MoClo kits for plants that work for both ... to share it with the scientific community! Good luck and happy cloning! This article was written by Susanna...

...binding. Suzuki et al. also envision eNLs as good reporter genes for endogenous protein fusions created using...Renilla luciferase variant fused to an FP; when supplied with a luciferase substrate, the luciferase transfers...Nano-lantern, a fusion of a mutagenized Renilla luciferase (RLuc8) with Venus. This luminescent technology is...Instead, a luciferase substrate like coelenterazine is supplied in the culture medium to allow RLuc8 to produce...constructs use the brightest characterized luciferase, NanoLuc, and its substrate furimazine. In addition...problems of low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently...transfer (BRET). Photons emitted by a Renilla luciferase variant are used to excite the fused fluorescent...

...Arabidopsis, and corn. For both β-glucuronidase (GUS) and GFP reporters, as well as multiple transient and...to be removed and replaced if needed. In total, three MCS regions permit multiple regulatory elements ...