...sgRNA targeting endogenous CD71 gene 46919 pMLS-SV40-EGFP Target EGFP gene that is stably integrated into...fold repress on targeted genes. 2. Two-plasmid CRISPRi system for mammalian gene knockdown The first plasmid... silence transcription on the gene expression level without genetically altering the target loci. Thus... pU6-sgGFP-NT1 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GFP (NT1)...binding platform, to repress expression of arbitrary genes in bacteria or human cells ( Qi et al. ). This CRISPR...designed with a 20-basepair complementary region to any gene of interest. Co-expression of dCas9 and sgRNA can...genome. 1. Two-plasmid CRISPRi system for bacterial gene knockdown The first plasmid ( pdCas9_bacteria ) ...

...reprogramming transgenes and the up-regulation of corresponding endogenous pluripotency genes. Retroviral...disrupt the expression of tumour suppressor genes and/or oncogenes, potentially leading to the presentation... Shao, L., & Wu, W. (2010). Gene-delivery systems for iPS cell generation. Expert opinion on biological...a single polycistronic vector or multiple single-gene vectors can be used to package reprogramming factors... the transposase. The donor plasmid contains the gene(s) of interest flanked by terminal repeat sequences...cassette comprised of OCT4, SOX2, LIN28, NANOG, and a GFP reporter. Minicircles are removed from cells by dilution...demethylation and expression of endogenous pluripotency genes (i.e. Oct4) are other measures of reprogramming....

...Express a gene of interest fused to the C-terminus of SCFP3A Amber N1 Yellow Mammalian Express a gene of interest...Express a gene of interest fused to the C-terminus of Amber mVenus N1 Yellow Mammalian Express a gene of interest...Clover (a GFP variant) commonly used with mRuby2 pLSSmOrange-N1 Orange Mammalian Express a gene of interest...biosensors targeting small molecules or specific genes are also available. Custom biosensors can also ...to create a fluorescent fusion protein with your gene of interest and the listed fluorescent protein. ...Aquamarine mCerulean N1 Cyan Mammalian Express a gene of interest fused to the N-terminus of monomeric...monomeric Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric...

...understanding of B. subtilis gene functions and interactions. The single-gene deletion library is derived...studying the phenotype of an essential gene, the CRISPRi Essential gene Knockdown library was constructed ...Bacillus subtilis libraries with Addgene – the B. subtilis Single Gene Deletion Library—Kanamycin and the...compatible cloning sites for easy cloning of your genes of interest. In order to improve the study of specific...cassette. However, it was unknown whether variations in gene expression from these polycistronic cassettes could... stoichiometry impacted reprogramming and global gene expression patterns. Polycistronic cassettes that...-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked...

...vectors for human gene therapy of glaucoma and their reversed cell entry characteristics. Gene Ther . 2020 ...157970 pTR-UF11 chimeric CMV/Chicken Beta actin (CBA) GFP Control Sergei Zolotukhin Citation Information AAV2...cells by a mutant adeno-associated viral vector. Hum Gene Ther Methods . Feb;25(1):72-82. PMID: 24191859 Ye...Adeno-Associated Virus Vector Expressing Retinoschisin. Hum Gene Ther Clin Dev . Sep;26(3):165-76. PMID: 26390090... Display Exceptional Characteristics for Retinal Gene Therapy. Mol Ther . Jun 3;28(6):1464-1478. PMID:...: Viral Vectors As part of our Viral Service , Addgene is producing ready-to-use viral preparations from...

...pAAV-CAG-FLEX-Archon1-KGC-EGFP-ER2 CAG Archon1 EGFP Cre dependent 5 Boyden 115892 pAAV-Syn-Archon1-KGC-GFP-ER2 Syn Archon1...Archon1 EGFP Constitutive 8 Boyden 115893 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] Syn Archon1 EGFP Cre dependent...HaloCaMP1a 138327 pAAV-synapsin-HaloCaMP1a-EGFP Syn HaloCaMP1a EGFP Constitutive 1, 9 Schreiter Calcium Sensor...HaloCaMP1b 138328 pAAV-synapsin-HaloCaMP1b-EGFP Syn HaloCaMP1b EGFP Constitutive 1, 9 Schreiter Dopamine Sensors...Ready-to-use AAV available from Addgene's viral service encoding biosensor tools, including calcium sensors...

...type or mutant ORFs, gene fragments, and gRNAs for genes linked to neurodegenerative disorders. Find plasmids...within the huntingtin (HTT) gene. An individual with the mutated HTT gene will have huntingtin proteins...association between a gene and late-onset AD. It is more likely a combination of gene variation and lifestyle... are associated with known genetic mutations in one of the following genes: SOD1, C9ORF72, TARDBP, FUS...three different inherited genes: Presenilin 1, Presenilin 2, and APP gene.The majority (>90%) of individuals...these additional genes may play in Alzheimer's disease. Research Tools Neurodegeneration Plasmid Collection...toxin receptor (DTR)–GFP fusion protein. Azim et al. Nature. 2014 Apr 17. Next-generation GRAB sensors for...

...plasmids allow cloning of protein coding sequences with GFP-LOVpep, cpPDZ, ePDZb and ePDZb1 as tags. pDawn/pDusk...pDusk Blue-light photoreceptor for light-controlled gene expression. These plasmids rely on the engineered... Science Guides Optogenetics Guide Optogenetics Guide Jump To: Optogenetics Guide Introduction Microbial...Collection . To find optogenetics plasmids available through Addgene, check out our Optogenetics Plasmid Collection...Addgene's guide to using Optogenetics plasmids in your lab, including explanations of microbial opsin...References Introduction The field of optogenetics integrates optics and genetic engineering to measure and manipulate...from Kir2.1 540 Plan Your Optogenetics Experiment When designing your optogenetics experiment, you’ll need...

...reporter gene (for example, luciferase or GFP) that offers a read-out of the activity of the genetic element...bacterial populations, an antibiotic resistance gene (i.e., a gene whose product confers resistance to ampicillin...transcription. Gene Knock-down Plasmids - Used for reducing the expression of an endogenous gene. This is frequently...chromosomes down to study their genes, marking the birth of molecular genetics. Today, scientists can easily...easily study and manipulate genes and other genetic elements using specifically engineered plasmids, commonly...when a gene is inserted within the MCS, its expression will be driven by the promoter. As a general rule...the gene, promoter, or other DNA fragment cloned into the MCS. The insert is typically the genetic element...

... this gene. Should you cut it at the beginning of the coding sequence to delete the entire gene? Perhaps... in vitro, and using it in mouse embryos to generate gene modified mouse lines. Hopefully this post will...they have been generated (more on that later). Find Validated gRNAs to Target Mouse Genes To begin the ...chances of gene modification in vivo. Find Plasmids for Testing Your gRNAs in vitro at Addgene CRISPR/Cas9...decide to do a gene modification experiment in mice to the day you have your F0 generation (barring life...Haoyi, et al. "One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated ...blogger Samantha Young. The use of CRISPR/Cas9 for gene editing has expanded since its adaptation for use...

...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002) and many ...2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters 520... acceptor GFP2-Gγ chimeras, and two untagged Gβ constructs. For the Gγ constructs, the GFP2 tag is fused... Gα-Rluc8 and Gγ-GFP2 will be in close association, resulting in a high level of GFP2 emission. Following... exhaustive refinement, it was not possible to generate an optimal Rluc8-Gα construct for two of the 16...interest, a Gα-Rluc8 plasmid, and the appropriate Gγ-GFP2 and Gβ constructs. Cells are then treated with the...complex will disassociate, resulting in a decreased GFP2 emission. The activation state can be measured by...

...6 letter representation of the gene (or DNA sequence). pBACKBONE-hGene Tip: Often researchers will add...pBACKBONE-Flag-hGene(Q295A)-EGFP  Wildtype version: pBACKBONE-Flag-hGene-EGFP or pBACKBONE-Flag-hGene(wt)-EGFP...would list it first. pBACKBONE-Flag-hGene If there was also an EGFP fused to the C-terminal of your insert...would list it after the insert. pBACKBONE-Flag-hGene-EGFP Other considerations  If your insert contains...be included in the plasmid name. Mutations are generally listed as the amino acid change and not a nucleotide...

...magnetic responses: EGFP, mScarlet, and AsLOV2. After several rounds of semi-random mutagenesis and screening.../nn.4265.  Additional resources on addgene.org Find Optogenetics Plasmids Find Biosensor Plasmids Visit... al., 2024). They deposited their plasmid with Addgene, hoping to inspire others to explore the phenomenon...Although a wide range of fluorescent proteins and optogenetic tools exist, humans (like many organisms) are...magnetoresponsive fluorescent protein. Controlling an optogenetic or fluorescent tool with magnets would enable... molecules like proteins with magnetic fields generally appears to be weak — if they exist at all. Andrew...protein they tested — E. coli expressing good ol’ EGFP — showed a slight but detectable change in fluorescence...

...comparing gene activation when enhancers were active or silent, they could identify the target genes of previously...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...updated Jun 24, 2020. Epigenetic modifications are an additional layer of control over gene expression that ...proteins that help package DNA, were associated with gene expression or silencing. To alter DNA modifications...oxidation and repair factors that allow for increased gene activation at the targeted site compared to other...the only CRISPR-based technology designed to alter gene expression. Fusing dCas9 to a transcriptional activator...a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers. Nature Biotechnology...

...include: Optogenetics AAV Control AAV Biosensor AAV And more! Optogenetics Optogenetics continues ...Nuc-flox(mCherry)-EGFP 27056 AAVrg pAAV-Ef1a-DIO EYFP 50457 AAV2 pAAV-hSyn-DIO-EGFP   Biosensor AAV...pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP 50457 AAV2 pAAV-hSyn-DIO-EGFP Recombinases Plasmid Serotype...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...you pick the best AAV for your experiment. The Addgene viral service has now expanded to offer many of... Plasmid Serotype Name 51502 AAV5 pCAG-FLEX-EGFP-WPRE 114471 AAV1, AAV5 pAAV-Ef1a-fDIO mCherry ...pEF1a-DIO-FLPo-WPRE-hGHpA    Additional resources on the Addgene blog Read all of our viral vectors blog posts ...

...AAV9 Optogenetics Adesnik New viral vector pAAV-nEF-Con/Foff DREADD Gi-mCherry AAV8 Chemogenetics Deisseroth... pENN.AAV.EF1a.eGFP.WPRE.rBG AAV PHP.eB Controls Wilson New serotype pAAV-CaMKIIa-EGFP AAV PHP.eB Controls...latest in Addgene's viral vectors services and resources!   More resources on the Addgene blog Addgene's...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share...Table 1: New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor...Deisseroth New viral vector pAAV.CMV.PI.EGFP.WPRE.bGH  AAV PHP.eB Controls Wilson New viral vector ...the full viral services collection or check out Addgene's Data Hub to view viral vector reports from other...

...victoria, AvGFP forms a homodimer. In corals, the red fluorescent proteins form tetramers. In general, fluorescent... develops genetically encoded FRET based biosensors.     Additional Resources on the Addgene Blog Which...detect weak dimerization tendency, as is present in EGFP. Constantini et al, (2012) developed a cell-based... properties of new fusions can be compared with mEGFP or mTurquoise2, which are established true monomeric...Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes. You can follow him... Protein Biosensors Additional Resources on Addgene.org Find Empty Backbones with Fluorescent Tags Find...

...for doing gene targeting, reporter generation, controllable gene knockouts, and inducible gene inactivation...Alec Kimmelman targets mouse metabolic genes with ~6 gRNAs per gene.  CRISPR-Act3.0, developed by Yiping... Shirley Liu's lab targets all mouse genes with 10 gRNAs per gene. The guides are optimized to maximize... to develop CRISPR-based approaches for studying gene function in these systems. As a part of this effort...Yiping Qi's lab, allows simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified...opportunities for investigating genetic mechanisms related to morphogenesis, cell fate specification, and...Lab have compiled a new toolkit for performing genetics studies in organoids. The Organoid EasyTag System...

...provide the Brunello (targeting human genes) and Brie (targeting mouse genes) pooled libraries from the labs...targeting EGFP, BRAF, and MAP3K4. The gRNAs targeting EGFP can be used as positive controls for gene editing...editing. You can also enhance expression of your gene of interest with a variety of Cas9 activators available...editing or gene activation, while the BRAF and MAP3K4 gRNAs allow you to target these widely studied signalling...pooled gRNA libraries that can be used to screen for genes involved in a wide variety of cellular processes...80,000 gRNAs to target around 20,000 individual genes. We provide enough lentivirus for you to both perform...and member of the Addgene Advisory Board) says: "When you're setting up to do a genetic screen, you need...

...promoter. Use this for doxycycline inducible (tet-off) gene expression! We've also added AAV1 and AAV5 preparations...variants of a new optogenetic tool called PdCO, deposited by the Yizhar lab. Addgene is now offering AAV1... pAAV_hSyn-SIO-stChrimsonR-EGFP-P2A-PdCO-miniWPRE AAV1, AAV5 Optogenetics Yizhar New viral service with...AAV1, AAV5 Optogenetics Yizhar New viral service with multiple serotypes pAAV_hSyn-PdCO-EGFP-WPRE AAV1... Brian O'Neill.  More resources on the Addgene blog Addgene's Data Hub What is a Virus (and a Viral Vector...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share...to our viral vectors repository since March.  Addgene is now offering an AAV2 CAG-driven tTA expresser...