...this is indicative of protein degradation). To control for protein degradation due to other pathways, ...electron microscopy analysis is time consuming. GFP-tagged Atg8p or LC3: ATG8 is part of a group of genes...with autophagosomal membranes, so tagging it with GFP allows for tracking the localization or accumulation...autophagosomes and autophagic bodies in yeast. A GFP-tagged version of LC3, the mammalian Atg8p homolog...pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic pHluorin (SEP). They are ...Fluorescent Protein Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy... Resources on Addgene.org Fluorescent Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors...

...experiment, consider co-transfecting with GFP. This allows you to sort for GFP-positive cells and to enrich for...articles (3/12/2014). With so many options for CRISPR plasmid tools and numerous experimental design decisions...Engineering Using CRISPR/Cas Systems has expanded out of control. To further consolidate these FAQs, Le Cong worked...genome editing using the Zhang lab's CRISPR/Cas plasmids. Designing your CRISPR genome editing experiment...with expression, though it is possible for the plasmid to still express without the G. Because the G is...marker to select transfected cells (for example, plasmid with a puromycin resistance cassette, such as PX459...related to the most frequently requested CRISPR plasmids at Addgene Find protocols and gRNA design tools...

...protein (Plasmid #141391-LV [+N]) or membrane protein (Plasmid #141386-LV [+M], negative control) were immunoblotted...of the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles.  ... over the original CROPseq. Find CROPseq-multi plasmids here Walton, R.T., Qin, Y., Blainey, P.C. (2024...potential improvements to the editor. Find PE7 plasmids here!     Figure 2: PE7 improves prime...also immunoblotted against vinculin as a loading control (right panel). Image from Addgene. These antibodies...conjugated with its HaloTag domain.  Find Voltron2 plasmids here! Abdelfattah, A.S., et al. (2023) Neuron...Figure 5: A) Representative confocal images showing GFP expression from ssAAV5s with hIBA1 or hIBA1a promoters...

...about GFP Controlling Protein Activity with GFP Resources at Addgene.org Find the Spinach Plasmids at ...nature of GFP fluorescence to create an improved fluorescent aptamer system. Knowing that GFP’s  structure...increase its fluorescence, essentially mimicking GFP.  Though the aptamer, called Spinach, required longer...at Addgene Find Broccoli Plasmids at Addgene View All Fluorescent Protein Resources ...

...PubMed. Find the plasmids from this publication at Addgene. Multichromatic control of gene expression... to neuroscience. In fact, precise light-based control of biological processes is highly useful in other... to be optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust...engineered to deliver precise, consistent, and controllable pulses of light. Complicated patterns of light...new pathways with sophisticated gene expression controls. Most cell types do not respond to low-intensity...two systems can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive...via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red light rapidly...

...with some plasmids. You just need three plasmids to start making AAV:  the packaging plasmid which contains... transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids have two ...of an AAV transfer plasmid. Parts shown in green are key elements of an AAV plasmid. Parts shown in grey... elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal ...AAV transgene. On a plasmid map, promoters are upstream or 5’ of the gene they control. Promoters can be...transfer plasmids have a promoter, but this element isn’t required when the transfer plasmid harbors non-coding...Transfer Plasmid video!   Now you’ve got a basic understanding of some common AAV plasmid elements!...

...and expression is controlled by the upstream recombination outcome. Find SPARC plasmids at Addgene To more...system along with precisely truncated attP sites to control recombination of the gene construct. A predictable...gene. Effector genes, any gene that you want to control the expression of using the SPARC system. The effector... more precisely control the amount of labeling, you can alter the attP site. Truncating the attP site ...effector position, as well as to optogenetically control the depolarization of R2 neurons using CsChrimson...transcription activation systems, such as LexA/p65 or mCD8/GFP, to allow sparse labeling in a variety of fly cell...genome engineering in flies Learn more about controlling gene expression in flies Resources on Addgene.org...

..., AAVrg Control Karl Deisseroth New serotype (multiple) rAAV-CAG::FLEX-rev:: hM4D-2a-GFP AAV1 Chemogenetics...pAAV-AiE0452h-minBG-SYFP2-WPRE3-BGHpA (Alias: CN2237) AAV-PHPeB Control Jonathan Ting New viral prep AiP12787 - pAAV-...minBG-SYFP2-WPRE3-BGHpA (Alias: CN2787) AAV-PHPeB Control Jonathan Ting New viral prep AiP12610 - pAAV-...pAAV-AiE0780m-minBG-SYFP2-WPRE3-BGHpA (Alias: CN2610) AAV-PHPeB Control Jonathan Ting New viral prep AiP13755: pAAV-AiE0743m...10aa-H2B-WPRE3-BGHpA (Alias: CN4496) AAV-PHPeB Control Jonathan Ting New viral prep AiP14825: pAAV-AiE0441h...prep pAAV_BiSSTe10_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New viral prep pAAV_BiSSTe10_ChR2... pAAV_BiCHATe27_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New viral prep pAAV_BiCHATe27_...

... create a fusion with a reporter protein such as GFP? All of these outcomes can be most effectively achieved...conjunction with activator or repressor domains to control gene expression. Base editors can help you make...add a tag, you will also need to deliver a donor plasmid or single-stranded DNA oligonucleotide, so you ...typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells or a selection...transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells. Likely contains...generate stable cell lines. Packaging and Envelope plasmids provide the necessary components to make lentiviral...for in vivo viral delivery. Cas9 mRNA and gRNA Plasmids containing gRNA and Cas9 are used in in vitro ...

...transformation Read our Plasmids 101 blog posts Resources on Addgene.org Browse our Plant Plasmids and Resources...page Find synthetic biology plant plasmids Find CRISPR plant plasmids ...its function. In nature Agrobacterium contains a plasmid with a T-DNA (transfer-DNA) region. During infection...check for GUS-like activity of your tissue. Run a control with Agrobacterium lacking the GUS gene since plants...promoter activity, for example. Useful positive controls could include: pSB96 - pL2_pSB90_2x35S::fLUC-I...tMAS::rLUC-I::pMAS. Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often a FP is fused to...localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different leaf...

... is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as...continue to add more controls, and have recently released these controls: pAAV-hSyn-DIO-EGFP (50457-AAV5): ...sensors Optogenetics Recombinases And more! Control AAV For many AAV experiments, we recommend validating...entire collection! We are expanding our entire control AAV inventory to give scientists access to a wide... Email us or check our website to access them! Controls pAAV-hSyn-hChR2(H134R)-EYFP (26973-AAVrg) pAAV-hSyn-hChR2...our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors... testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This...

...conjugative mobilization of broad host-range IncQ plasmids." Plasmid 62.2 (2009): 57-70. PubMed PMID: 19465049...page Deposit microbiology plasmids with Addgene Find synthetic biology plasmids ...family-RP4 conjugation system. This system can deliver plasmids to both Gram-positive and Gram-negative cells.... Wang lab further developed a modular suite of plasmids (pGT) with various replicative origins that can...selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both sorted using FACS and isolated...genera across four phyla that have taken up the plasmid through conjugation in vivo studies. However, they... tools for microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to...

...protein control. We also added the toxin-antitoxin stability cassette Axe-Txe so that the plasmid could ...accessory genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient transfection... 1248 ng of CMV-gvpA-IRES-BFP plasmid and 954 ng CMV-gvpNJKFGWV plasmid into 70%-confluent well of HEK...in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe (Fig. 1a).  With pBAD-bARGSer-AxeTxe... Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram of pBAD-bARGSer-AxeTxe. (b) Methods to ...overexpressed compared to accessory proteins. Excess gvpA plasmid added into the transfection mixture in HEK293T ...mammalian species to produce gas vesicles. New mARG plasmids Our new mARGs come as a set of two PiggyBac integratable...

...Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET ... No spatiotemporal control. Many PPI detection methods lack spatiotemporal control or an “on/off switch...partners if they are enriched relative to controls using Split-BioID-GFP and BioID run on six unrelated proteins... by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent biotinylation...method Enter Split-BioID, a spatiotemporally controllable version of the BioID protocol. The critical ...BirA* fusion proteins are expressed in cells via plasmids or by the creation of stable cell lines and these...methods, including BioID, is its spatiotemporal controlled labeling of interacting proteins; nearby proteins...

...In a previous post from our Plasmids 101 series, we learned how the Cre-loxP recombination system can... even in the presence of Cre recombinase. This plasmid now specifically drives expression of mCherry instead... should contain a reporter DNA sequence (such as GFP) in the sense orientation, followed by coding sequence...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ensure...et al. Nat Biotech 2003): In the presence of Cre, GFP expression is lost, allowing for rescued expression...2286602. PubMed Central PMCID: PMC3406316. Find FLEx plasmids from these Addgene depositors: Ed Boyden Bernardo... plasmids...

...you find what you need.   More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular...function of Your Gene Of Interest (YGOI) in a controlled environment under various conditions. The first...need. You will also have to make sure that your plasmid has been incorporated into the host organism, usually...great for overexpressing YGOI in an isolated, controlled environment. In most cases bacterial expression...open reading frame (ORF) of interest –  try pRS plasmid shuttle vectors. You may prefer the more traditional...using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following the expression pattern...details and guidance for choosing your perfect plasmid backbone. Or just ask –  leave a message in the...

...Fluorescent Proteins Control Protein Expression with Cre-lox Control Protein Activity with GFP Resources at ... the plasmids from Zhou et al here. In addition, it could be possible to use pdDronpa to control other...transmitted by intermediate messengers. Optical control of kinase activity can provide greater spatiotemporal...There is no common method for creating optically controlled kinases. The Solution: In Zhou et al, the Lin... the delayed effects because you can precisely control the timing of kinase activity. 2. Studying negative...photoswitchable kinases in vivo not only allows for precise control of a signaling pathway in the context of a whole...Pengpeng Li, Kang Shen, and Michael Z. Lin. "Optical control of cell signaling by single-chain photoswitchable...

...without mCherry and GFP fluorescent controls. Alternatively, you can order individual plasmids from the library...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome...streamline identification. While the Zhang lab controlled their multiplicity of infection so that a single...disease depends on both genetic and expression control systems to mimic illness on a molecular level. ... of the collection, we’re releasing individual plasmids in batches. If there’s something you’d like that...

...antibodies against popular fluorescent proteins (GFP and YFP) and protein tags (c-MYC and HA). We have...materials, repositories provide standardized quality-control and technical support to the community. Thanks ...about Scientific Sharing: The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years Data...

...curated collections of plasmids as kits with greatly reduced costs per plasmid. We generally create kits...just the tool you need. The plasmids in this portion of the BIOFAB Plasmid Set contain terminators of ...populations. This kit consists of a 48 plasmid adapter library, with each plasmid carrying a unique barcoded Illumina-M13F...researcher at a nonprofit institution, you can order plasmids covering a wide range of fields from Addgene. ...kits to allow you to easily use a set of plasmids together on a tight budget. To help popularize our ...order from the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit... production (a variety of pExpression vectors) plasmids. The type IIS restriction sites used in these ...