... time or location can be critical when, say, the gene is essential during development and a knockout is...activity can be monitored by phosphatidic acid probe GFP-PASS. B) Time series of HEK 293T cells expressing...membrane-targeted SuperPLDx30 (magenta) and PA probe GFP-PASS (green). After 30 minutes of blue light illumination...the Addgene blog Viral Vectors 101: Optogenetics Additional resources on Addgene.org Optogenetics Guide... The Chow Lab has also generated a variety of membrane-recruitment optogenetic tools for manipulating ...a full toolbox and workflow for generating new Rho GTPase optogenetic systems (Berlew et al., 2022). Stopping...LOV, CRY2-CIB, and other optogenetic modules? Or want to find more optogenetic tools in our collection?...

...translational. Ex: the discovery of GFP. Chapter 4: Goals of the Center for Regenerative Meidince - Understanding...Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids from the...treat disease through tissue repair, stem cells, gene therapy, and tissue engineering. How the multidisciplinary...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...Darrell Kotton is the director of the Center for Regenerative Medicine at Boston University Medical School...source biology in his own lab, at the Center for Regenerative Medicine, and within the biomedical research...Along these lines, Darrell recently joined the Addgene Board of Directors where he hopes to help keep ...

...NIR FPs can be used similar to GFP-like probes by supplying a single gene to cells. Still in bacteria, ..., which do not produce BV, an additional gene coding for heme oxygenase should be supplied (Baloban et...crosstalk-free three-color imaging with green and red GFP-like FPs (Shcherbakova et al., 2016), as well as ...applications of fluorescent proteins, genetically encoded biosensors, and optogenetic tools. Follow her recent research...-the-art probes, many of which you can find at Addgene. Biliverdin chromophore Near-infrared fluorescent...FPs can be combined with blue-light controlled optogenetics, such as channelrhodopsins and tools based on...reported widely used dimeric iRFPs (Table 1). In general, the monomeric state of a probe leads to less interference...

...ideal for large gene transfer. The M24 promoter provides constitutive, high-level gene expression and ...further customizing gene expression. Sahoo et al. have tested the system with genes up to 4 kb in length...versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants...Scientists have worked to optimize this system for gene transfer, studying the stability of modified Ti plasmids...versatile binary Ti vector for both transient and stable gene expression applications in plants. This smaller,...and many plant scientists want to transfer large genes, smaller binary vectors with a larger cloning capacity.... They also replaced the promoter regulating the gene of interest with a constitutive, robust M24 promoter...

...proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes are expressed under native...sequence and a selectable gene product for isolation of successful transformants. Addgene distributes several...be interested in attaching epitope tags to their genes of interest, allowing for easy capture and detection...modifications in S. Pombe, including full and partial gene deletion, overexpression (by promoter substitution...either the N- or C-terminus (3xHA, 13xMyc, GST, or GFP). Longtine et al.(3) describe a complimentary set...few. Have you used these tools in your own lab? Addgene would love to hear from you, our community, about...

... Gerlich 25999 LV-GFP Chromatin H2B GFP Elaine Fuchs 11680 H2B-GFP Chromatin H2B GFP Geoff Wahl 21210 ...61803 GFP-Rab7A Late endosomes Rab7a AcGFP Gia Voeltz 12605 GFP-rab7 WT Late endosomes RAB7 GFP Richard...Targeting Gene/Signal Fluorescent Protein PI 110060 FUW mCherry-GFP-LC3 Autophagosome LC3 mCherry, GFP Anne...Gadella 50057 pLYS1-FLAG-MitoGFP-HA Mitochondria MCU GFP Vamsi Mootha 49153 GFP-Mff Mitochondria-Outer ...Targeting Gene/Signal Fluorescent Protein PI 26722 GFP-cortactin Actin Filaments Cortactin EGFP Anna Huttenlocher...89472 GFP-hChibby1 Mother Centrioles / Ciliary Base Chibby1 GFP Ken-Ichi Takemaru 89770 pLenti-EGFP-hChibby1...Filaments beta-actin EGFP Robert Singer 27382 pDEST/N1-hEB1-GFP Microtubules EB1 GFP Robin Shaw 40908 pDEST...

... PINK1 N-GFP PINK1 GFP CMV Parkinson's Mark Cookson 13316 pcDNA-DEST47 PINK1 C-GFP PINK1 GFP CMV Parkinson's...21190 GFP-pcDNA3-PKCgamma-cys1Acys1B PRKCG GFP CMV Spinocerebellar ataxia 27 Tobias Meyer 21204 GFP-N2-PKCgamma...PKCgamma PRKCG GFP CMV Spinocerebellar ataxia 26 Tobias Meyer 21205 GFP-C1-PKCgamma-C1A PRKCG GFP CMV Spinocerebellar...-HtrA2-EGFP HTRA2 GFP CMV Parkinson's L. Miguel Martins 14122 pcDNA3-HtrA2-EGFP S306A HTRA2 GFP CMV Parkinson's... GPD 25QDProGFP p416 HTT GFP GPD Huntington's Susan Lindquist 15569 GPD 104QDProGFP p416 HTT GFP GPD Huntington's...GAL 25Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15581 GAL 46Q+ProGFPp416 HTT GFP GAL1 Huntington's...GAL 72Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15583 GAL 103Q+ProGFPp416 HTT GFP GAL1 Huntington's...

...biosensors To focus on a particular gene or protein, check out our gene-specific biosensors. These constructs...biosensors as an example. Many calcium biosensors include GFP fused to calmodulin (CaM) and the chicken myosin ...Visualization Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent... to the molecule it detects. This change then generates a reporter signal. Reporter signals may be electrochemical... OR enzyme activity. Small molecule biosensors Addgene has many sensors that measure the concentration..., and voltage across membranes. These sensors generally follow the same pattern described above - a sensing...fluorescent molecule   Additional Resources on the Addgene Blog Check out the Rosella Autophagy Sensor Read...

...attach the gene of interest to either the N-or the C-terminus. Even if a researcher’s gene of interest...from Joachim Goedhart: Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection.  ...researchers as we could, and Addgene is a wonderful vehicle for us to do that.” At Addgene, we are equally excited...is available in an easily searchable format on Addgene’s Michael Davidson Fluorescent Protein Collection...collection with over 300 backbones alone,” said Addgene’s Lianna Swanson, who has been working with members...sun, from the standard oldies but goodies (e.g., EGFP and YFP) to the new and improved fruit colors (e.g...the scientific community," says Joanne Kamens, Addgene's Executive Director, "and to promote the great ...

... for gene expression GFP Varies pBABE GFP - Retroviral gene expression pMKO.1 GFP - Retroviral...pPS808 - N-terminal GFP for yeast expression pET GFP - C-terminal GFP for...expression MSCV-IRES-GFP - Mammalian retroviral gene expression with GFP selectable ...C elegans expression GFP Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression... from the SP6 promoter pCS2 mt-GFP - Create myc-tag, GFP-tag fusions for transcription ...expression pcDNA3 GFP LIC cloning vector - C-terminal GFP for mammalian expression...vector for gene expression (tet inducible) Zeocin/Bleo Mammalian pLenti X1 GFP-Zeo DEST ...

...cells - one containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3) and one with either tTA...the env gene containing the rev response element (RRE), an internal promoter, and the desired gene (transfer...-1 genome containing all viral genes with the exception of the env gene (packaging plasmid), and 3) the...used to express your gene of interest either constitutively or inducibly. If your gene of interest is toxic...expression of your gene. The addition of Dox in the culture will then repress your gene of interest. This... Youichi Suzuki (2011). Gene Regulatable Lentiviral Vector System, Viral Gene Therapy, Dr. Ke Xu (Ed.)...harmful genes into a transfer vector. Lentiviral particles expressing proto-oncogenes or oncogenes for instance...

...48201 CAG-GFP-IRES-CRE Cre and GFP coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG...116879 CAG-Cremyc-2A-GFP GFP and Cre CAG Mammalian Lu 117148 Hiv7CMV-Cremyc-2A-GFP GFP and Cre CMV Lentiviral...pCALNL-GFP Cre dependent GFP expression Mammalian Cepko 8389 p212 pCMV-EGFP/RFP EGFP-dsRed gene switch...iRFP670-EFS:Cre-2A-GFP iRFP670, Cre, and GFP TRE Mammalian Jacks 68544 AAV-Cre-GFP Cre CMV AAV Nestler...shRNA expression. Gene Switch: These constructs contain two genes of interest, Genes A and B. When Cre...promoterless CRE-GFP fusion none Mammalian Sauer 11960 pBS537 tet-hCMV-GFPcre tet inducible Cre-GFP fusion tet-hCMV... CMV Lentiviral Fuchs 26646 pCAG-Cre-IRES2-GFP Cre and GFP coexpression CAG Mammalian Chenn 26647 pCAG-Cre...

...blot showing GFP expression in cells that have been transfected with a plasmid encoding GFP. Actin, a common...which come from naturally-occurring immunoglobulin genes, recombinant antibodies are plasmid-based. Their... What is an Antibody? Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies...cascades that clear these infections. Anything that generates an antibody response in the immune system is referred...in the DNA of the hybridoma can occur, known as genetic drift, meaning that over time, the antibody you...monoclonal antibodies, recombinant antibodies are a homogeneous population. However, unlike monoclonal antibodies...antigen binding, the variable regions alone can be generated as a fusion protein. An ScFv is made up of the...

...both introduce genes into an organism’s genome (see figure) and to disrupt endogenous genes at the site ...first-generation transposase (as determined by transposition assays of antibiotic-resistance genes conducted...). In their Nature Genetics paper, the lab showed that efficiencies of stable gene transfer using SB100x... the rapid generation of stable transgenic cell lines which can robustly express your gene-of-interest... exciting tool for producing specific genetic mutations and gene disruptions without the use of viral ...transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun;41(6):753-61. Epub 2009... et al. A new approach to gene therapy using Sleeping Beauty to genetically modify clinical-grade T cells...

... in E3 and insertion of GFP expression cassette. Bunz 179202 pAd5-B6/7 ∆E3-GFP Adenoviral A block for ...regions present in pAd5-B6 ∆E3-GFP and pAd5-B7 . Bunz 179203 pAd5-B1-deltaE1-GFP Adenoviral A block for the...containing transgene under CMV promoter Vogelstein 16404 pAdTrack Shuttle For production of GFP-trackable...containing transgene under a chosen promoter Vogelstein 16405 pAdTrack-CMV Shuttle For production of GFP-trackable...GFP-trackable viruses containing transgene under CMV promoter Vogelstein 16407 pAdEasy 2-GFP beta-gal Shuttle Test...adenoviral plasmids. Want an adenoviral plasmid with your gene of interest? Search our database for adenoviral ...with large inserts Vogelstein 179201 pAd5-B6 ∆E3-GFP Adenoviral A block for the AdenoBuilder genome assembly...

...read-through transcripts of the effector gene. Effector genes, any gene that you want to control the expression...chemical inducers of gene expression (del Valle Rodríguez et al., 2011).  Genetics of SPARC Figure...Learn more about controlling gene expression in flies Resources on Addgene.org Find fluorescent protein...truncated attP sites to control recombination of the gene construct. A predictable number of cells will express...binds to the UAS binding motif, driving downstream gene expression. attP and attB, recombination sites. ...side of a stop cassette, upstream of an effector gene. In cells where the stop cassette is removed during...expression of either a stop cassette or an effector gene. HDVR (2X Hepatitis Delta Virus Ribozyme), a self-cleaving...

...typically include a gene that is advantageous for survival, such as an antibiotic resistance gene. In contrast...the target gene. Vital component for expression vectors: determines which cell types the gene is expressed...scientists use plasmids? Generally, scientists use plasmids to manipulate gene expression in target cells...replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing bacteria...MCS is often downstream from a promoter. Insert Gene, promoter or other DNA fragment cloned into the ...obtained. Selectable Marker The antibiotic resistance gene allows for selection in bacteria. However, many ...referred to as “vectors” or “constructs.” To insert a gene of interest into a vector, scientists may utilize...

...CRISPR-Cas9 constructs from Addgene to gene-edit mutations into XP genes and other genes involved in photosensitivity...researchers identified the gene responsible for the disease as ATP1A3. The gene encodes a sodium-potassium...Syndrome, a devastating genetic disease caused by mutations in the OCRL1 gene that leads to the early ...studies on OCRL1 already underway in his lab. The gene appears to play a role in many cell processes involving... learned there was a disease associated with the gene, it piqued their interest. They obtained cells from... University with David Goldstein’s lab, will use GFP-ATP1A3, FLAG-ATP1A3, and ATP1B3 plasmids to explore...McSweeney says they now think that mutations in the gene may prevent neurons from returning to basal conditions...

...this plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY...already have a pBAD33-based plasmid expressing your gene of interest, all you have to do is co-transform ...co-transform pBLADEONLY. That way, a previously cloned gene of interest, under the pBAD promoter, can be controlled...strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter. They ... Resources on Addgene.org Check out the Molecular Biology Reference Download Addgene's Plasmids 101 eBook...simply more precise. “We show that you have less heterogeneity in the population and you can do all of these...permission.   BLADE is just one of the several optogenetic tools that Di Ventura’s lab and Khammash's lab...

...reporter like GFP, some smaller CRISPR-Cas proteins such as SaCas9, CRISPR gRNAs, or your gene of interest...through this challenging sequence. Transgene The transgene is the gene that will be delivered by the AAV...expression of the AAV transgene. On a plasmid map, promoters are upstream or 5’ of the gene they control. Promoters...plasmid which contains the structural and packaging genes, the adenoviral helper plasmid which contains the...for the successful and specific expression of your gene of interest. References Haery L, Deverman BE, ...Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods 30:206–213 . https://doi.org/10.1089...control transgene expression. Lox sites are directional 34 bp sequences that can flank the transgene and ...