...be paired with Cre-expressing transgenic mouse lines to restrict transgene expression to specific tissues...control transgene expression. Gene regulatory elements [promoters and enhancers, miRNA target sites, and...2021) can help restrict expression to your cell type of interest, while miRNA target sites can be used...optogenetic and chemogenetic tools. Dual expression Promoters, miRNAs, enhancers, and switches all take up...determine cell type- or region-specific transgene expression. (a) Several engineered capsids with unique tropisms...specificity. An on/off genetic switch can be used for expression control. (c) Viral delivery methods include focused...efficiency in the brain along with high off-target expression, particularly in the liver, when administered...

...Addgene.org Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati...catheter, tail vein and femoral cut down injections. Journal of Biosciences and Medicines, 08(09), 131–141. ...

....org Check out Our Hot Plasmids Articles to See Compiled Info on New Plasmids Review the CRISPR Field...casually mention “Haery et al., showed that MYC expression was increased…” when discussing the review topic...Field with Our CRISPR Guide Pages ...was invited to write on a particular topic. Alternatively, maybe you’re researching a topic for which ...article doesn’t turn into a textbook. Get the journal’s submission rules for review articles Whether submitting... you have these rules (word limit, formatting guidelines, etc.) you have some criteria to shape the document...easily convert the citation style to whatever the journal requires. Using the (Last name, Year) format also...

...plant plasmid resources at Addgene  Choosing an expression system Choosing an appropriate expression vector....org Find plant plasmids Visit our plant resource page Deposit arabidopsis plasmids with Addgene ...., 2007). Alternatively, if you prefer Golden Gate assembly, there are a series of plasmids from the Dinneny...this method include: Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a...options, but here are a couple of my favorites. Plasmids from the Tsuyoshi Nakagawa lab are binary gateway...for visual selection of transformants based on expression of RFP in the seed (Emami, Yee and Dinneny, 2013...culture you are growing definitely contains your plasmid. Grow lots of Agrobacterium: The more agrobacterium...

...their seminal work on micro RNAs in C. elegans. The C. elegans work on small RNAs informed and drove work ...modular toolkits build single plasmid targeting vectors that encode the gRNA transcript and the repair template...step as simple as heat-shocking to induce the expression of a Cre recombinase contained within the knock-in... C. elegans, visit Addgene’s C. elegans CRISPR plasmids page. Future perspectives It is interesting to...using modified Cas9 proteins to regulate gene expression (CRISPRi/CRISPRa), visualize specific genomic...Addgene's CRISPR Resources Learn about gRNA design and gRNA optimization for your CRISPR experiment ...depending on perspective – to work with inefficient sgRNAs in my initial direct screening efforts, which were...

...freezer, you’ll probably find lots of different plasmids containing the same protein of interest tagged... whatever molecule you choose (Figure 1). Just express the fusion protein and add a specific ligand! A...better detect dim structures or low levels of expression with the same imaging workflows you already use...Addgene’s collection, so you may be able to get a plasmid ready-to-use without any cloning at all! As mentioned...Dyeing Through Chemistry Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands GCE4All...10.1038/s41467-023-39655-3 Wilhelm, J., Kühn, S., Tarnawski, M., Gotthard, G., Tünnermann, J., Tänzer, T.,...

...Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor plasmid contains 1 kb homology...mutations in crRNA target sites found within the homology areas of the plasmid to prevent the plasmid from being... the tag. Created with BioRender.com. Plasmids, plasmids everywhere To find out more about gene edited...CRISPR Tagging Plasmids Browse the Fluorescent Protein Collection Check out the CRISPR Guide Pages .../Cas9 (Cas9 protein and the crRNA-tracrRNA complex) and the donor plasmid into the cells we use electroporation...Further, the Yamamoto PITCh system plasmids provides an alternative to HDR as the double-strand break ...expensive for many labs. We’ve created and tested plasmids that use CRISPR/Cas9 to endogenously tag a wide...

...population. It can be used to measure protein expression, identify rare cells, or even sort out single...using the location of the cell populations as a guide  (Figure 3.) You can see that in our dot plot, we... Conventional vs. Flow Cytometry Plasmid-based Recombinant Monoclonal Antibodies Introduction to Antibodies...PLOS ONE, 12, e0180664. https://doi.org/10.1371/journal.pone.0180664 Boix, F., Llorente, S., Eguía, J., Gonzalez-Martinez...infection in liver and kidney recipients. World Journal of Transplantation, 8, 23–37. https://doi.org/10.5500...PloS One, 10, e0137550. https://doi.org/10.1371/journal.pone.0137550 More resources on the Addgene blog Conventional...

...transfer plasmid), the envelope (Env) gene on a separate plasmid (envelope plasmid), and the polymerase ...different plasmids. For a retroviral system this looks like the genetic cargo in one plasmid (the transfer...Gag structural protein on yet a third plasmid (packaging plasmid). Viral production then proceeds by transfection...Addgene viral services Addgene Biosafety Guide Addgene Viral Plasmids Which Type of Virus Should I Use? ... transfection of all three plasmids into a producer cell line, such as HEK-293T, production of viral particles... and Pol genes are not encoded in the transfer plasmid, the component that physically integrates into ... retroviral packaging components onto distinct plasmids for safety.    Viral cargo-specific safety...

...advantage of DNA sequencing to turn mAb clones into expression constructs to be conveniently modified and engineered...atlas will enable scientists to have an intuitive guide for checking their staining and neuroanatomy without...Biol 13(6): e1002165. https://doi.org/10.1371/journal.pbio.1002165 More resources on the Addgene blog Antibodies...Antibodies 101: ValidationSelecting the Right AntibodyPlasmid-based Recombinant Antibodies    ...

...quantification of mRNA, which can be used as a proxy measure for differences in protein expression. (If you're...you can assume that qPCR can accurately detect expression differences of twofold or greater with good statistical...shown to have as much as sixfold variation in expression levels reported (Biassoni, 2014), so you’ll want...Housekeeping Genes as Reference for qPCR Gene Expression Analysis During iPS Reprogramming Process. Sci...; Higgins PJ; Crawford DR. Control selection for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7...Germain H. Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable...blood glucose-regulated and can up-regulate the expression of GAPDH. Biochem Biophys Res Commun. 2001 Mar...

...depositing plasmids pre-publication Find molecular biology protocols Find eBooks on plasmids, CRISPR, viral...changes in root-to-shoot ratio in Arabidopsis, (2) expression patterns in plants with enhanced sodium accumulation...scientific journal. The peer-review is a service that we, as scientists, provide for journal editors to...been peer-reviewing journal submissions since 2015 for a wide variety of journals. Since then, I developed...the submitted paper in addition to the journal-specific guidelines.  Peer-review step 1: The first read ...whether work is suitable for publication in their journal. The early peer-review attempts took place at the...submission, then for the submission to the subsequent journal.  As a postdoctoral researcher studying plant development...

...Resources Online Microscopy Primer on the Molecular Expressions Website Other major microscopy manufacturers ...PubMed PMID: 12624904. 5. Power RM, Huisken J. A guide to light-sheet fluorescence microscopy for multiscale... Find Empty Fluorescent Protein Backbones Find Plasmids for Optogenetics ...photon microscopes. Total Internal Reflection Microscopy (TIRF) Total Internal Reflection Microscopy (TIRF... are the main focus of the experiment, a Total Internal Reflection Fluorescence (TIRF) microscope can ...2015). PubMed PMID: 25998828. 4. Axelrod D. Total internal reflection fluorescence microscopy in cell biology...

...PMCID: PMC3810607. 2. Bajar, Bryce T., et al. "A Guide to Fluorescent Protein FRET Pairs." Sensors 16.9... create fusions with a gene of interest. These plasmids will be most useful for constructing intermolecular...including fluorescent proteins, has led to the alternative acronym Fluorescence Resonance Energy Transfer... endogenous state that you want to measure (overexpressing a gene or protein after transfection). Future...stress sensor for specific proteins in situ." FEBS journal 275.12 (2008): 3072-3087. PubMed PMID: 18479457...Biosensors and Intravital FRET Microscopy." Biophysical Journal 111.6 (2016): 1103-1111. PubMed PMID: 27475975....