...viral particle) and toxicity of the gene product (i.e., if the gene encoded by the transfer plasmid is ...hUbC-driven EGFP; can be used for cDNA expression Baltimore 12247 pLVTHM 2nd EF-1a-driven GFP and shRNA ...promoter for transgene and CMV drives expression of RFP as a reporter. See plasmid 17618 for GFP plasmid. ...24129 pULTRA 3rd bi-cistronic expression of EGFP and the gene of interest, also can express shRNA (cloning... which can be modified to encode a wide range of gene products. The main restrictions on a lentiviral ...chimeric 5’LTR, Any expression cassette (promoter and gene of interest) can be cloned into the plasmid. Nolan...gateway plasmid, constitutive Hygromycin resistance gene coexpression. See article for other versions of ...

... Cas9 (dCas9) fused to a fluorescent marker like GFP, researchers have turned dCas9 into a customizable... throughout the chromosome. Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!...

...listed below. When targeting other gene loci, you will prepare the gene-specific CRISPR and donor vectors...FLP expression vectors, a general cloning plasmid, and a prebuilt Unc-32::GFP targeting vector. Jorgensen...multiple genes. Plasmids can be found associated with the following article: Paix, et al. Genetics 2014 Allen...CRISPR plasmids for endogenous tagging of your gene of interest. CRISPR...repair template design. How to use CRISPR to tag your gene of interest Mendenhall and Myers Tagging System ... the plasmids as listed in each row. If your own gene of interest is currently unavailable, you will need...terminal affinity tag (3xFLAG-2xSTREP) on endogenous genes for the isolation of native protein complexes. This...

...Root 105533 pAAV.CMV.Luc.IRES.EGFP.SV40 Firefly CMV AAV expression of firefly luciferase and GFP James Wilson... choice in assays ranging from use as a reporter gene in vitro and in vivo to use in biosensors. The most...effect of mutations of these regulatory elements on gene expression. Empty backbones for inserting regulatory...an intron in the middle of the Firefly luciferase gene.. TREAT (3( T hree)′- R NA E nd A ccumulation during...into which you can clone your regulatory element or gene of interest into to create a luciferase reporter... Firefly/Renilla Creating ORF-Firefly luciferase gene fusions. Renilla luciferase under the control of...enhancer regions. Mutations within the luciferase gene allow for increased copy number in E. coli and expression...

...possible deletion strategies for genes and non-coding elements. For creating a gene knockout, two sgRNA located...evaluate the RNA, perform RT-qPCR for gene expression of the relevant gene 7,8 . To evaluate the protein, perform...genome editing tool that allows genetic perturbation of genes and genetic elements. Here we present a simple... as pX458 (Addgene plasmid ID 48138) or pX459 (Addgene plasmid ID 48139), which include GFP and puromycin...for efficient loss-of-function studies of genes and genetic elements in mammalian cell lines. Protocol...CRISPR/Cas9 construct (10 μg total). Add 0.5 μg of GFP expression construct. Electroporate cells with 250...filter into a FACS tube. FACS sort the top ~3% of GFP positive cells in order to enrich for cells that ...

...reporters simultaneously. By fusing sfGFP and/or mCherry to genes of interest, it is thus possible to ...to bind on the chromosome. When dCas9 binds to a gene, it blocks the progression of the RNA polymerase...enables robust, noiseless knockdown of bacterial genes. Molecular Systems Biology, 14(3):e7899. doi: 10.15252... CRISPR knockdown to modulate expression of two genes without cross‐talk. (A) Schematic of the strain ... the tracrRNA sequence (not shown). (B) Relative GFP and RFP concentration given relatively to the non‐targeting...carrying two CRISPR guides. The first one binds to sfgfp with either 0, 10, 11, 14, or 20 matching nucleotides...sites. The levels of the two reporters, mCherry and sfgfp in this case, can be controlled using a plasmid‐...

...popular markers like tubulin or epitope tags like Myc, GFP, and more. Neuroscience : Antibodies targeting proteins...Browse Addgene's collection of recombinant monoclonal antibodies. All...application-specific validation and quality control by Addgene as well as by our trusted partner labs. On this...antibodies currently available from our catalog. Addgene values transparency and accessibility in research...reactivity. Reactivity is based on validation data from Addgene or trusted partner labs or is predicted based on... on sequence homology with the antigen. Addgene supplies a list of recommended applications based on our...immunohistochemistry, and immunocytochemistry. Addgene will continue to work with both depositing and ...

...reporter gene (e.g. GFP) to identify and enrich positive cells, or selection marker to generate stable ...knockin. Select gene and genetic element to be manipulated In order to manipulate a given gene using CRISPR...mutant allele of a gene ( point mutant )? Increase or decrease expression of a target gene? Once you have ...separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich...sequence for the gene you are trying to target. However, the exact region of the gene you target will ...want to: Generate complete and permanent loss of gene expression or function ( knockout )? Generate a specific... Edit Generate a specific user-defined sequence change in a particular gene, such as generating a point...

...vector Retroviral vectors: neomycin (pSIR-neo) , GFP (pSIR-GFP) , DsRed (pSIR-DsRed-Express2) , human CD2 (...long or short list of genes provided by the user. Read more about it on the Addgene blog . Eukaryotic Pathogen...identify positively or negatively selected sgRNAs, genes, or pathways. CrispyCrunch CrispyCrunch is used ... genomes (200+) that can predict gRNA targets in gene families, HDR oligonucleotide design, and batch ...sgRNA design tool for genome editing, as well as gene regulation (repression and activation). Genome support...target sites by off-target quality. Validated for gene inactivation, NHEJ, and HDR. Reference genomes include... to identify novel gRNA target sites in a single gene, as well as a target site common to a set of similar...

... pCAG-SpCas9-GFP-U6-gRNA 79144 Mammalian hU6 yes, cut S. pyogenes EGFP Zou pCAG-eCas9-GFP-U6-gRNA 79145...Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA(F+E) 59986...cut Lb Cpf1 Neo Welker AIO-GFP 74119 Mammalian U6x2 yes, nick S. pyogenes EGFP Jackson AIO-mCherry 74120...are expressed as polycistronic glycine tRNA-gRNA genes (PTGs) and then PTG cassettes are assembled into...capable of simultaneously repressing multiple target genes in Escherichia coli. Koffas Do you have suggestions...none S. pyogenes Zhang pL-CRISPR.EFS.GFP 57818 Mammalian/Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert...none S. pyogenes Zhang pLKO5.sgRNA.EFS.GFP 57822 Mammalian/Lentiviral BsmBI none S. pyogenes EGFP Ebert ...

...protein, permeabilized cells  Cy5NTAc 32 25-45 GFP (fluorescein is similar)   Lumio Red Tc: CCPGCC 48...in setting up a FRET-based assay or screen (the Addgene blog has an excellent primer on FRET and how to...Fluorescent proteins (FPs) are the gold standard for genetically-encoded FRET donors (Fig. 1). Scores of FP variants...presented (Fig. 1) which can be carried out using Addgene plasmids. His-tag labeling reagents You may already...Gradia Lab has deposited a gateway vector with Addgene, where you can insert your protein of interest,...PubMed Central PMCID: PMC4380784. Resources on the Addgene Blog Get Tips for Using FRET in Your Experiments...Your Fluorescent Protein Basics Resources on Addgene.org Find Plasmids for FRET Browse Empty Plasmids...

...labeling. Size is also a factor. Bulkier tags, such as GFP, may interfere with protein expression or function...the dye and the endogenous target, and thus no genetically encoded tag is required. These dyes can also ...be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that covalently reacts...maintained through mechanisms like fission, fusion, biogenesis, and mitophagy, can be tracked using dyes like...Keller, P. J., Ji, N., & Lavis, L. D. (2017). A general method to fine-tune fluorophores for live-cell ....1016/j.bbamem.2013.09.005 More resources from Addgene Fluorescent protein plasmids and resourcesA practical...

.../Tag Serotype(s) PI 105551 pENN.AAV.CamKII.HI.GFP-Cre.WPRE.SV40 CamKII GFP 9, rg* Wilson 105558 pENN.AAV.CamKII...site-specific recombinases, which can be used to control gene expression. Dre AAV ID Name Promoter Fluorophore...2, 5, 8, 9, rh10 Wilson 105545 pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 CMV eGFP 1, 2, 5, 8, 9 Wilson 55632 pAAV-Ef1a-mCherry-IRES-Cre...EBFP-Cre Syn EBFP 5 Zeng 105540 pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 Syn eGFP 1, 2, 5, 8, 9, rg*, PHPeB Wilson...

...reporter gene (e.g. GFP) to identify and enrich positive cells, or selection marker to generate stable ...knockin. Select gene and genetic element to be manipulated In order to manipulate a given gene using CRISPR...mutant allele of a gene ( point mutant )? Increase or decrease expression of a target gene? Once you have ...separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich...sequence for the gene you are trying to target. However, the exact region of the gene you target will ...want to: Generate complete and permanent loss of gene expression or function ( knockout )? Generate a specific... Edit Generate a specific user-defined sequence change in a particular gene, such as generating a point...

...another marker, such as GFP. A researcher can use FACS to sort cells expressing GFP and later expand these...Available online: Genetic Organization and Genetic & Structural Features . In vivo gene delivery and stable...transfer plasmid. 2nd generation lentiviral plasmids utilize the viral LTR promoter for gene expression, whereas... rev, and tat genes all from a single packaging plasmid such as psPAX2 . 3rd generation packaging systems...packaging plasmid encoding the Gag, Pol, Rev, and Tat genes. The transfer plasmid contains the viral LTRs and...pictured). Unless an internal promoter is provided, gene expression is driven by the 5'LTR, which is a weak...: Uses 3 separate plasmids encoding various HIV genes. Safer. Replication incompetent and always SIN: ...

...experiment, consider co-transfecting with GFP. This allows you to sort for GFP-positive cells and to enrich for...example, choosing target sites that span the knock-in gene. For making mutations, one good way is to mutate...be okay if you are introducing a selection marker gene. The homology arms should be no more than 100bp ...for my genomic PCR, but couldn't amplify the EMX1 gene using same primer you used in the Science paper ...You may also like... Zhang Lab CRISPR plasmids General CRISPR Plasmids Dr. Le Cong, the first author of...additional PAM sequences should not be sufficient to generate off-target DSBs; there needs to be extensive homology...below are considerations for designing HR donor. Generally if off-target (non-specific) cleavage is not a...

...find antibody plasmids for: Common antigens such as GFP or mCherry Monoclonals, Nanobodies, Sybodies, or ...-mouse" or "anti-rabbit") ID Plasmid Description Gene/Insert PI Return to top Do you have suggestions ...Find information on Addgene's collection of plasmid-based antibodies, including monoclonal antibodies...Plasmid Collection Antibody Plasmid Collection Addgene distributes ready-to-use recombinant antibodies...application-specific validation and quality control by Addgene as well as by our trusted partner labs. Learn More...animals or hybridoma cells, but are difficult to generate due to expense and time. An alternative approach...peptide, from the popular SunTag system, fused to sfGFP for imaging. Learn more about antibodies and their...

...briggsae , etc.) gene or sequence: ID Plasmid Gene/Insert PI Don’t see your gene listed? Know of some great...can knock down genes by feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily... used in worms, search Addgene's entire collection. ID Plasmid Description Gene/Insert PI Plasmids Expressing...which diminishes epigenetic silencing in the germline. A "FLP-Out" system for controlled gene expression in...Expressing Worm Genes/Inserts The table below lists plasmids that contain a worm ( C. elegans, C. briggsae...vectors for C. elegans research, including lacZ and GFP fusion vectors. C. elegans optimized fluorophores...Plasmids for the temporal and spatial control of gene expression in C. elegans by combining expression...

... not endogenously-expressed (e.g., using an anti-GFP antibody), the target antigen is first transiently...distributed by Addgene have been produced in-house by Addgene scientists. They are generated using standard...or by contacting help@addgene.org . Some of the antibodies distributed by Addgene were developed by the...Overview of Addgene's antibody production methods and quality control processes for purified recombinant...optimized for each specific antibody in order to generate high quality preparations. Following production...titered and undergo stringent quality control by Addgene or a trusted quality control partner before distribution... preparations developed at IPI may differ from Addgene antibodies in “Production & Usage” and “Delivery...

...measurable through an epitope tag such as FLAG/His/GFP. Fluorescent protein tags, which correlate closely...procedure and application to the major ALS/FTD disease gene C9ORF72. eLife, 8, e48363. https://doi.org/10.7554...101: Validation Resources on addgene.org Addgene's Antibody Guide Addgene's Antibody Protocols... outlined below. Knockout cell line approach Genetic knockout-based validation can conclusively prove...you are working with cells that are amenable to genetic knockout, this represents a very robust approach...to start with a screening procedure in easily genetically modified cell types (top). The specific approach...10.1038/s41598-019-55133-7 More resources from the Addgene blog Introduction to Gating in Flow Cytometry Antibodies...