...exhibiting its versatile utility in quantifying multiple gene editing activities. Click here to find the ACE ...Wanisch  Listen to the optogenetics podcast segment LOVTRAP is an optogenetic system that was recently... the University of North Carolina (plasmids at Addgene). It consists of two domains, LOV2 and Zdk1, where...function is more specific and much faster than any genetic or pharmacological manipulations on cells, and ... riboswitches used in the Riboglow platform at Addgene. Braselmann E, et al. Nature Chemical Biology...real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry fluorescence...involves tracking the ratio of mCherry-positive to eGFP positive cells through fluorescence microscopy or...

...ampicillin resistance gene, you’ll find the full name of the gene, the name of its gene product, and a description... plasmid features such as antibiotic resistance genes, selectable markers, promoters, origins, tags, and... Overview of SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid...powered by GSL Biotech's SnapGene Server Software. With the backing of SnapGene’s sequence viewer software...the features you need quickly. SnapGene’s Feature Library - SnapGene’s software seamlessly identifies ...nucleotides, such as the fluorescent proteins EGFP and mEGFP. Many of these features, such as Cas9, weren...scientists) for suggesting these changes, and the SnapGene and Addgene software development teams for working tirelessly...

...been used for many years for in vivo delivery and gene therapy and represent one of the most studied viruses...CRISPR-Cas. Figure 1: Schematic of targeted gene knock-out (left) and targeted DNA knock-in (right...binds to target DNA. In the final step of targeted gene knock-out (left), a fragment is deleted from two... This capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral...Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an insertion, a deletion or a frameshift...with Cas9 and a gRNA designed to target a mutated gene. In a Scientific Reports paper introducing the delivery... CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various cell types...

...could replace one of the reporter genes above with your gene or genes of interest. Some of the plasmids...reporter gene). Addgene's collection offers a variety of plasmids to express two or more genes, some of... that are expressing your gene, especially if you are studying a novel gene, is not always a straightforward... your gene of interest, scientists have instead developed novel methods to co-express your gene along ...with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to easily screen...more of the genes replaced.  Additionally, 2A peptides could be PCR-cloned between your genes of interest...more than two genes. Plasmid Name Multicistronic Element Expression Type MSCV-IRES-EGFP IRES Retroviral...

... what part of a gene enhancers like to interact with, and how many promoters or genes each enhancer is...plot line in gene regulation. When chromatin conformation data is compared with changes in gene expression...dynamic role in gene regulation The high resolution and multiplexed chromatin ‘photos’ generated with CAPTURE...changes in gene expression and determine who’s a key driver of these changes. By generating a more vivid...updated Jul 2, 2020. Promoters may be the star of gene regulation, but enhancers and chromatin looping ... bound by transcription factors, can increase a gene’s transcription. Sometimes enhancers are located ...located thousands of base pairs away from the gene they regulate, but are brought in proximity by the looping...

...luciferase or eGFP) flanked by the EBOV-specific gene start and gene end sequences. Thus, the minigenome...replication and viral gene transcription (6). The EBOV genome contains seven viral genes, each flanked by ...sequence (gene start) and a transcription termination/polyadenylation signal sequence (gene end). A minigenome...Figure 1). All viral genes are removed and replaced by a single (non-viral) reporter gene (e.g. firefly luciferase...Cloning the viral genes in this plasmid would be more difficult compared to if each gene were encoded in...transcribe the reporter gene (e.g. firefly luciferase). The polymerase recognizes the gene start signal and ...of the reporter gene. The polymerase will elongate the mRNA until it reaches the gene end signal, where...

...observed in mammalian cells is to tag an endogenous gene. By imaging cells (or tissues) that produce an endogenous...fluorescent proteins or using endogenous tagged genes) can be used to verify performance under different... develops genetically encoded FRET based biosensors.     Additional Resources on the Addgene Blog Which... QY (EC*QY, sometimes normalized to the value of EGFP). The higher the number, the higher the theoretical...mVenus as a reference protein is available from Addgene. Another way to dodge cellular variation observed...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...

... Figure 1: In conventional gene targeting (A), you must first generate your desired mutation in mouse...mutations, the 8-10 month process of conventional gene targeting is lowered to about 2-3 months. But certain...example of inserting a 2A-FlpO cassette into the Fgf8 gene. Miura et al. (2018) suggest using a 50-80 bp sequence... mouse model generation easi-er! Generating mice before CRISPR The process of generating knock-in mice...process of mouse model generation simpler, as described in an earlier post on generating CRISPR mouse models...after editing. How to use Addgene plasmids for Easi-CRISPR You can use Addgene plasmids to create a repair...template. Figure 2: Overview of ivTRT to generate an ssDNA repair template. Although effective...

...excellent series of posts by Joachim Goedhart on the Addgene blog – outlining various important considerations...relating to that specific protein (for instance: EGFP).  Users can “favorite” certain proteins, or create...github @tlambert03.   Additional Resources on the Addgene Blog Fluorescence Microscopy Techniques - Which...Proteins for Multicolor Imaging Resources on Addgene.org Browse all fluorescent protein plasmids Find...

... editing multiple genes at once; using dual nickases to generate a knockout or gene edit; or using Cas9...the targeted gene. The ideal result is a loss-of-function mutation within the targeted gene. However, the...eliminate target gene function. It has also been used extensively to screen for novel genes that regulate... activate, or repress genes. Each library typically contains ∼3–6 gRNAs per gene to ensure modification...repress, visualize, and isolate genes. Activation or Repression of Target Genes Figure 9: Overview of CRISPRi...highest levels of single-gene activation (Figure 9D) In bacteria, activating gene expression is more difficult... of effective gene activators when fused with dCas9. Recently, synthetic CRISPR-Cas gene activators have...

...Entrez Gene matches to your insert. Select the appropriate gene if it appears in the Entrez Gene list, ...correct Entrez Gene does not appear in the auto-populated list, try entering the gene's Official Full ...known as" aliases in the Gene/Insert box. Once you have selected the appropriate gene from the list, save ...bacterial strain(s) through Addgene. Description – please indicate the gene cloned into your plasmid, any... for the gene or insert. Step 4: Growth and Distribution Please indicate which strain Addgene should propagate... Encodes insert(s) A plasmid with one or more genes, protein coding sequences, or other elements inserted...Empty backbone A plasmid designed for cloning a gene/insert into it. Empty backbones often contain epitope...

...remove genes flanked by either loxP or frt sites respectively, and empty vectors to express your gene of ...and puromycin resistance gene from the CMV promoter pCDH-CMV 72265 Express gene of interest from the CMV...pCDH-EF1 72266 Express gene of interest from the EF-1 promoter pCDH-CB 72267 Express gene of interest from ...promoter pCDH-EF1-copGFP-T2A-Puro 72263 Express copGFP and the puromycin resistance gene from the EF1 promoter...the cloning capacity. pCDH-CB-IRES-copGFP-T2A-Puro 72299 Express gene of interest from the CB promoter ... 72484 Express gene of interest from a truncated EF-1 promoter pCDH-EF1-Luc2-P2A-copGFP 72485 Expresses...Kazuhiro Oka from the Baylor College of Medicine Gene Vector Core Viral...

... Endogenous Gene Tagging in Human Cells on the Addgene Blog. Plasmids for Endogenous Gene Tagging in Human...Link opens in a new window) RNP transfection for gene editing human iPS cells (Link opens in a new window...87421 TUBA1B-mEGFP AICSDP-4 mEGFP Alpha-tubulin Microtubules 87422 LMNB1-mEGFP AICSDP-10 mEGFP Lamin B1 Nuclear...envelope 87423 TOMM20-mEGFP AICSDP-8 mEGFP Tom20 Mitochondria 87424 DSP-mEGFP AICSDP-9 mEGFP Desmoplakin Desmosomes...87425 ACTB-mEGFP AICSDP-15 mEGFP Beta-actin Actin filaments 87426 SEC61B-mEGFP AICSDP-7 mEGFP Sec61 beta...101782 LAMP1-mEGFP AICSDP-19 mEGFP LAMP-1 Lysosome 101783 MAP1LC3B-mEGFP AICSDP-25 mEGFP Autophagy-related...101786 ST6GAL1-mEGFP AICSDP-26 mEGFP Sialyltransferase 1 Golgi 109122 NPM1-mEGFP AICSDP-50 mEGFP Nucleophosmin...

...in neurons. (b) Genetic cargo can be customized to control transgene expression. Gene regulatory elements... relatively even distribution of target gene expression (EGFP, in the depositing manuscript) across the....     Figure 1: AAV toolkit for targeted gene delivery to the brain. The AAV (a) capsid, (b) cargo...Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory element; NHP, non-human primate. ...Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nature Protocols;14, 379–414...2017). Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems...Gradinaru, V. (2022). Engineered AAVs for non-invasive gene delivery to rodent and non-human primate nervous...

...MiniPromoter Source Gene Construct Reporter Negative Control N/A pEMS1312 cre N/A pEMS1301 cre/EGFP/NLS N/A pEMS1308... disorders by enabling region- and cell-specific gene-delivery in the mouse brain through these human ...pEMS1308 EGFP/cre N/A pEMS1302 EGFP/cre/NLS N/A pEMS1306 EGFP/NLS N/A pEMS1307 EGFP/NLS N/A pEMS1313 intron-lacZ...pEMS1153 EGFP/NLS Ple37 CRH pEMS1154 EGFP/NLS Ple38 CRH pEMS1155 EGFP/NLS Ple39 CRH pEMS1156 EGFP/NLS Ple44...pEMS1113 EGFP/cre/NLS Ple51 DBH pEMS1362 EGFP/NLS Ple52 DCX pEMS1198 EGFP/NLS Ple53 DCX pEMS1199 EGFP/NLS ...pEMS1167 EGFP/NLS Ple63 DRD1 pEMS1168 EGFP/NLS Ple64 FEV pEMS1129 EGFP/cre/NLS Ple64 FEV pEMS1398 EGFP/NLS ...pEMS1160 EGFP/NLS Ple93 GPR88 pEMS1161 EGFP/NLS Ple94 GPR88 pEMS1162 EGFP/NLS Ple95 GPR88 pEMS1163 EGFP/NLS...

...each gene. Click the gene name for a full list of plasmids available. Please note that some genes may ...Plasmids by Gene The following table lists genes in the ARF GAP and GEF family, information on each gene, and...following table lists genes in the ARF GTPase family, information on each gene, and examples of plasmid types...have plasmids available for request. Family Gene Entrez Gene ID Length (aa) Example Expression Systems ...this page was generated with the help of Richard Kahn . Browse ARF GTPase Plasmids by Gene The following...

...G-deleted rabies lacks the viral glycoprotein (G) gene, which is not required for the transcription or .... Since this starter cell now has all the viral genes present, the virus can spread from this cell to ...Ready-to-use AAV available from Addgene's viral service encoding tools for monosynpatic retrograde neuronal...to perform retrograde neuronal tracing. Note : Addgene does not distribute rabies virus. AAV Vectors for...Description Serotype PI 52473 pAAV-synP-FLEX-splitTVA-EGFP-B19G Can be used to complement deletion-mutant rabies...virus AAV1 Wickersham 100798 pAAV-syn-FLEX-splitTVA-EGFP-tTA These AAV together can be used to complement... was modified so that it could only infect a genetically specified neuronal population. This is done using...

...mCherry and firefly luciferase. pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro - Lentiviral vector...Addgene's collection of plasmids for viral pseudotyping with SARS-CoV-2 spike protein. Collections...the live virus. Alternatively, it is possible to generate pseudovirus, where the structural and functional...protein and you will find examples of each in Addgene's collection: HIV-based lentiviral particles Murine...particles Vesicular stomatitis virus particles See Addgene's Viral Plasmids and Resources page for more information...lists expression plasmids that are suitable for generating pseudotyped virus for SARS-CoV-2 and other coronaviruses...vector expressing destabilized EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing firefly...

...top Plasmids Encoding Mammalian Genes or Inserts Several mammalian genes have been identified as having...to these genes below. ID Plasmid Description Industry PI For more information on these genes, see the ...Cas12a plasmids Addgene's full CRISPR guide and summary of CRISPR pooled libraries . FlipGFP-based activity...Addgene's collection of plasmids and resources available for Coronavirus research. Collections...related coronaviruses. To assist with this effort Addgene will maintain this plasmid collection page, which... Kostman for the Innovative Genomics Institute Addgene COVID-19 Resources The following types of resources...mammalian sequences linked to SARS-CoV-2 infection. General Tools : Plasmids expressing Cas12a/Cas13a, Reverse...

... editing multiple genes at once; using dual nickases to generate a knockout or gene edit; or using Cas9...the targeted gene. The ideal result is a loss-of-function mutation within the targeted gene. However, the...eliminate target gene function. It has also been used extensively to screen for novel genes that regulate... activate, or repress genes. Each library typically contains ∼3–6 gRNAs per gene to ensure modification...repress, visualize, and isolate genes. Activation or Repression of Target Genes Figure 9: Overview of CRISPRi...highest levels of single-gene activation (Figure 9D) In bacteria, activating gene expression is more difficult... of effective gene activators when fused with dCas9. Recently, synthetic CRISPR-Cas gene activators have...