...expression plasmid. Experimental use Since the initial report describing CRISPR-mediated purification of a specific...including Protein A, 1xFLAG, 2xAM, and HA have been reported, as well as affinity purification with an anti-Cas9...chromatin immunoprecipitation (enChIP)." Scientific reports 3 (2013): 3171. PubMed PMID: 24201379. PubMed Central...Find CRISPR plasmids by function Find recently deposited plasmids ...

...are removed and replaced by a single (non-viral) reporter gene (e.g. firefly luciferase or eGFP) flanked...necessary to direct minigenome replication and reporter gene transcription by the EBOV polymerase (reviewed...the minigenome as a template to transcribe the reporter gene (e.g. firefly luciferase). The polymerase...start signal and initiates transcription of the reporter gene. The polymerase will elongate the mRNA until...

...technology more user-friendly and precise. We will deposit the plasmids of these systems to Addgene in the...Scientific Community Addgene: In 2012 your lab first deposited plasmids with Addgene. In 2014, you joined Addgene's..., as well as the Editorial Board of Scientific Reports and PLoS One. Additionally, you are involved in...shopping for a wide range of plasmids. The merit of depositing plasmids to Addgene has been spectacular. We'...

...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...

...-founded by Harvard Investigators and Addgene depositors David Liu, PhD, Keith Joung, PhD, and Feng Zhang...Center will continue to expand as Rett scientists deposit their MECP2-related plasmid tools with Addgene ...Researchers can also find links to cell line repositories including patient-derived iPSCs, MECP2 animal...Neurological Disorder by Programmable RNA Editing. Cell Reports 32:107878 . https://doi.org/10.1016/j.celrep.2020.107878...

... extract as much information as possible from depositing laboratories and associated publications, but...would ‘spot-check’ critical regions of plasmids deposited with us via Sanger sequencing, while more recently...Generation Sequencing (NGS) technology. As our repository continues to expand, we realized the need to ...scale our lab work as we grew. As the number of deposited plasmids increased, the accompanying sequencing...

...pooled library.  Pooled libraries Scientists have deposited many types of pooled libraries here at Addgene... from the original library constructed by the depositing lab. (For our more popular libraries, you might...recommend following the protocols provided by the depositing lab whenever available. Addgene also shares the... other information or scripts provided by the depositing lab for data analysis.  I'm often asked whether...

... (Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing) technique Sensitivity: Single molecule...DNA polymerase, recombinase, and the ssDNA-FQ reporters that exhibit fluorescence upon cleavage. Upon ... results in the trans cleavage of the ssDNA-FQ reporter resulting in fluorescence. Cas12a binds all the...results in the continuous cleavage of the ssDNA-FQ reporter. The resulting fluorescence can be observed under...

...in the same lane, and the resulting fraction is reported. Figure 2: Normalization with loading controls...equally strong, so the normalized value simply reports which POI band is stronger. The right gel has exactly...abundance on your membrane, which would be directly reported by band intensity. Researchers use later steps...perturbed by the experiment Unfortunately, multiple reports suggest that common housekeeping proteins are influenced...

...integrin antibodies available through Addgene’s repository. The collection includes 26 unique antibodies...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... by IPI. Our integrin antibodies are the first deposit from an expanding collection of readily accessible...

...of interest and co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence localizes to the plasma...And orthogonal sensors with different ITAM/SH2 reporter pairs allow multiplexed measurement of different...10.1101/2022.08.13.503850.  iGECInano NIR calcium reporter Near-infrared (NIR) fluorescent proteins promise...photochemical characteristics than previous NIR calcium reporters iGECI and NIR-GECO2G, and its small size (~1500...

...stage-specific promoter and design a UAS reporter construct, the reporter will only be expressed where and/or...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...

...that uniquely express selectable and/or sortable reporters in certain states of interest. For example, in...above (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 ...

...approaches may become even more powerful following a report that one can use linear repair templates (ie. PCR-derived... 2014). A further development of this approach reported in 2017 uses the direct injection of a CRISPR/...templates (Farboud et al., 2019).   The Mello lab also reported that modifying the 5’ modification of oligos used...mixtures (Doksin et al., 2018). The Mello lab also reported efficient editing using a co-injection marker ...

...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...really cool MoClo kit, we would love for you to deposit it with us to share it with the scientific community...Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media...

...protein analysis products. You can find plasmids deposited by Promega here.    References and resources ...ligands for dopamine D2/D3 receptors. Scientific Reports, 10(1). https://doi.org/10.1038/s41598-020-78827...fluorescence-based ligand-binding assay. Scientific Reports, 6. https://doi.org/10.1038/srep25769 Grimm, J....mitochondrial dynamics in mammalian cells. Biophysics Reports, 3(4–6), 64–72. https://doi.org/10.1007/s41048-...

...and a ssDNA-fluorescently quenched reporter is added. As this reporter is degraded by Cas12a, it will release... using a fluorescent reporter. In SHERLOCK, a quenched ssRNA fluorescent reporter is added to the reaction...following the amplification protocol specified by the depositing lab. After amplification, you’ll need to use ... target sequence, it promiscuously cleaves the reporter as well, and a quantifiable signal is produced...CRISPR 101 eBook (3rd edition) CRISPR software and depositor protocols JOVE video reprint: Protocol for genomic...NNNNGATT Streptococcus thermophilus (ST) 3' NNAGAAW Treponema denticola (TD) 3' NAAAAC Additional Cas9s from...all-in-one CRISPR/Cas9 vector system. Scientific Reports , 4 (1). PMID: 24954249 Shechner, D. M., Hacisuleyman...

...LaFleur, who shared information about recent CRISPR deposits from Arlene Sharpe’s lab. X-Chime and Performing...Faulds, who introduced the very first antibody deposit (the Integrin collection) from the Institute for... Thirteen: Guest Contributors of 2023 8 Great Depositor Research Stories Interested in contributing to...

...www.addgene.org/deposit or email deposit@addgene.org for more information or to start a deposit. If you have...

...telling scientists about Addgene’s repository, and explaining how to deposit and request plasmids, but I also...for: The inside scoop from scientists who have deposited plasmids at Addgene.  Learn how discoveries were...