...DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your insert of interest...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3...1.3 pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...volume 500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI ratios...

... of our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression... Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), is also available in AAV1. Plasmid Serotype .... Find the Dolcetto lentiviral prep here Control AAV Control AAV allow researchers to not only interpret...mediated neuronal control. The advantages of optogenetics lie in the spatiotemporal control offered by the...respectively of a GFP dependent Cre. The two segments come together in the presence of GFP to form a functional... pAAV-hDlx-GqDREADD-dTomato-Fishell-4 Controls Plasmid Serotype Name 114471 AAV2 pAAV-Ef1a-fDIO...mCherry 83900 AAVrg pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg pAAV-mDlx-GCaMP6f-Fishell...

...GBPs/GFP. The new system, CRE-DOG (Cre Dependent On GFP), is activated by GFP and derivatives GFP and ...to bind GFP. The idea of using GFP as a scaffold suddenly seemed very realistic. Envisioning GFP as a substrate...presence of GFP. These systems, known as T-DDOG and Cre-DOG, respectively, repurpose popular GFP reporter...lines expressing GFP in specific cell types to do more than just label cells. If GFP could be co-opted...co-opted to control gene expression, he could then selectively manipulate only GFP-labeled cells. Once Tang ...tested pairs of their GFP-binding proteins (GBPs) to find those that could co-occupy GFP. Once they’d found...culture system. In cells cotransfected with GFP and the GBPs, GFP linked the two GBPs together to create a...

...scientists to begin to study GFP and deduce its biochemical properties. Now GFP and its fluorescent derivatives...biology. Scientists can easily utilize GFP-containing plasmids as a means to many functional ends. We’...fusions. Transcription Reporter: Placing GFP under the control of a promoter of interest can be used to...selection: Expression constructs like plasmids often include GFP as a marker to help identify which cells...taken up the plasmid. This can serve as an alternative to antibiotic selection. Plasmids of this type ...type may have the GFP under the control of an additional promoter from that of the gene of interest, or expressed...expressing GFP from cells that are not. Developmental/transgenic uses: Because of its stability, GFP can be...

...those formed from control (GFP gRNA-targeted) BRAF sites (lanes 3, 4) were used as controls. Duplexes were...from BRAF-edited and GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification...amplification products (GFP-GFP duplex) as a control. 4. After optimizing the reaction and confirming the presence...express CAs9 and are included as a control (WT). Vinculin is a loading control. 3. In terms of making the... for gene editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011, 59702-LV) was used to assay... by the virally-delivered GFP gRNA, will edit out the virally-delivered GFP gene. So, cells with no Cas9...I thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low...

...popular control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs ...about. Controls Plasmid Serotype Name 37825  AAVrg (20 µL size serotype testing) pAAV-CAG-GFP 50465... Plasmid Serotype Name 105530  AAV1 pAAV.CMV.PI.EGFP.WPRE.bGH 37825  AAV1 pAAV-CAG-GFP 59462  AAV2...Chemogenetics Plasmid Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype...intersectional studies And more! Control AAV We’re always excited to add new control AAVs to our inventory since...optogenetics inhibitors Plasmid Serotype Name 99039  AAVrg pAAV-CamKII-ArchT-GFP (PV2527) 105679  AAV5... testing) pAAV-CAG-GFP 50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The...

...pAAV.hSyn-FLEX.iGABASnFR.F102G pAAV-Syn-Archon1-KGC-GFP-ER2 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] pAAV-FLEX-tdTomato pAAV-GFAP104...pAAV-mDlx-NLS-mRuby2 CAG-NLS-GFP pAAV-hSyn-mCherry Read on to learn more!  Calcium sensor AAV The GFP-based calcium...pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2]) of this membrane-embedding...serotype you’d like to be notified about. Controls Plasmid Serotype Name 114472  AAV5, AAV8  pAAV-...indicators Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST Chemogenetics Plasmid Serotype...37825  AAV8*, AAV9*  pAAV-CAG-GFP 50465  AAV8*, AAV9*  pAAV-hSyn-EGFP *Also coming in our new 20 ...cre-dependent version at Addgene! Fluorescent protein and control AAV These fluorescent protein expression vectors...

...the various controls that can be used for plasmid-based experiments. What are control plasmids? Generally...Negative controls: untreated cells, empty vector control, and non-targeting control Negative control conditions...follow an example of a plasmid-based experiment, delineate its requisite control plasmids, and discuss why ...Transfection controls: empty vector and internal control A transfection control measures transfection efficiency...cells. One transfection control is an empty vector control; specifically, the plasmid without the independent...empty control vector would be Plasmid A sans shRNA, or backbone Y alone. The empty vector control allows...internal control may be a plasmid that constitutively expresses a reporter protein (e.g., GFP or luciferase...

...decrease in red puncta. Find the mRFP-GFP-LC3 plasmid at Addgene! GFP-LC3-RFP-LC3∆G: A fluorescent probe ...an internal control. Autophagic flux is estimated by the GFP/RFP ratio: a decrease in GFP fluorescence...Addgene! mRFP-GFP-LC3 tandem fluorescent protein This autophagy biosensor contains mRFP and GFP fused to the...flux. mRFP-GFP-LC3 functions as an autophagy bionsensor via the same mechanism as Rosella: GFP is quenched... that releases an internal control Another autophagy biosensor is the GFP-LC3-RFP-LC3ΔG probe. It’s similar...molecule of GFP-tagged LC3 for every one molecule of RFP-tagged LC3∆G. Like the unmodified LC3, GFP-LC3 is ...decrease in autophagic flux.   Find the GFP-LC3-RFP-LC3∆G plasmid at Addgene! References Klionsky, Daniel...

...Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor plasmid contains 1 kb homology...and tested plasmids that use CRISPR/Cas9 to endogenously tag a wide variety of genes with GFP. These plasmids...found within the homology areas of the plasmid to prevent the plasmid from being cut and destroyed by CRISPR...introduced donor plasmid as a repair template. This results in the insertion of the GFP tag precisely where... the tag. Created with BioRender.com. Plasmids, plasmids everywhere To find out more about gene edited...individual proteins under endogenous regulatory control. But even with the advent of CRISPR/Cas9 technology... in human cell lines. Furthermore, the quality control necessary to ensure the edited cells are behaving...

...Chang-Deng Hu Green sfGFP Split super-folder GFP, our most-requested GFP(1-10) and GFP(11) Versatile protein...Commun. 2016 Bo Huang spGFP Split superpositive GFP Split-superpositive GFP reassembly is a fast, efficient...popular split-FP plasmids from our catalog to help you find the right split-FP plasmid for your experiment...live cells.  Controls  When designing experiments, it’s important to include proper controls to assess the...base FP (for example, EBFP2(1-10) can be used with GFP(11)). Some of the articles linked below only provide...below, and click the Article link to find all the plasmids from the associated paper:   Color Base FP...Blue EBFP2 EBFP2(1-10) and Capri(1-10) for use with GFP(11) Multiplexed labeling of cellular proteins with...

...light-based control of signaling pathways. This is particularly exciting because using light to control signaling...  Option #2 may sound like fiction, but light-controlled protein purification is now a reality thanks ..., and their use for protein purification and controlling cell signaling. Key components of OptoNBs OptoNBs...Lab monitored how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target proteins...co-localization with a membrane-localized mCherry or GFP protein. With this system, if the OptoNB can bind...insertion into loops 1 and 6 resulted in light controlled binding, but with opposite effects: insertion...more OptoNBs, one against mCherry and one against GFP. Insertion of AsLOV2 into loop 1 always generated...

...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...subunit (RPP30) as a control for normalization purposes are being used. Quality Control Mycoplasma Our 293T.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...virus preps are titered and subjected to quality control by Addgene before being distributed to customers...production protocols, titering methods, and quality control are described below. AAV Production AAV distributed...probes targeting the ITR elements and an internal control of known titer (protocol modified from Lock et ...

...expression plasmid encoding the ORF to be expressed Finally, the researcher needs a plasmid encoding their...TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can be...be used as a positive control during optimization experiments. This version of GFP will contain a phosphoserine...-phosphorylated or ‘wild type’ control protein. The ‘wild type’ control can be made one of two ways. First...using the same expression plasmid. This is a particularly important control when creating a phosphoserine-incorporation...Sep-OTSλ plasmid - Addgene # 68292 C321.ΔA bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG GFP plasmid...Find Synthetic Biology Plasmids Find Bacterial Expression Systems Find Plasmids for Protein Purification...

...released Anti-GFP [N86/8R] as another option for targeting GFP. But why might you need a second GFP antibody...western blot for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP [N86/8R] page...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ...  Find heat-inducible lentiviral plasmids here!   Wu Y, et. al. Control of the activity of CAR-T cells ...PMCID: PMC9015817.   Meet anti-GFP [N86/8R]! by: Ashley Waldron Anti-GFP [N86/38.1R] was among the first... missed something smokin'? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ...system Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder YFP ...

... an inducible promoter (Plasmid 64542 or 64543) Before translation, both GFP and RFP are bound to the...Addgene.org Find Fluorescent Protein Plasmids Check Out Other Hot Plasmid Technologies ...researchers explore the complexity of translational control in cellular physiology. Ribosome profiling Ribosome...switching, TRICK requires the following components: a GFP protein that binds to the coding region of a reporter...reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter...appear yellow. The act of translation pushes NLS-PCP-GFP off of the mRNA, leaving only a red FP bound to the... contains a nuclear localization signal, NLS-PCP-GFP then returns to the nucleus to bind cognate untranslated...

...(cap) expressing plasmids is generated. Diversity is introduced into the cap plasmid library by inserting... comparison, capsids were packed with a GFP transgene and GFP intensity and transduction efficiency of...Caltech stepped up to the challenge. Find the PHP Plasmids Here! Their Solution: Use the CREATE method to...region of the cap gene. In this special CREATE cap plasmid, loxP sites flank the downstream polyA sequence...sequence. When viruses produced from this plasmid pool are injected into an animal with cell-type-specific Cre... do these AAVs transduce the CNS and PNS? Both plasmids infect the PNS or the CNS much better than the...AAV.PHP.eB and AAV.PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were derived...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...Science Find the Cas14 plasmids Anti-CRISPR proteins for optogenetic control of CRISPR-Cas9 Anti-CRISPR... Find the anti-CRISPR plasmids Tagging human transcription factors with GFP Kevin White’s lab recently...deposited a set of CRISPR plasmids to tag human transcription factors with GFP. As part of the Encyclopedia...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Read the paper in Science Find the new SHERLOCK plasmids More precision with dual-nuclease Cas9-Cas9 ...Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit for genome editing in plants...

...recombinase. If GFP is under control of an endogenous promoter, you can use expression GFP to track cells...tag an endogenous protein with GFP, as seen in blue flame plasmid OCT4-eGFP-PGK-Puro from the Jaenisch lab...knockout a given gene and insert GFP into the genome, we’d create a plasmid similar to the one shown below...designed to insert GFP into a given locus.   Conditional knockouts The methods and plasmids described in...DNA designed in the lab. Specially constructed plasmids can be used to replace genes in yeast, mice, or...gene. Here, we'll describe the techniques and the plasmids used to inactivate specific genes in mammalian...mind when choosing a knockout strategy! Knockout plasmids Homologous recombination is a mechanism to accurately...

...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...and GFP shRNA. Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...inducible inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al...editing. Dr. Masahito Ikawa has created a GFP reporter plasmid for scientists to validate the efficacy ... target sequence, the plasmid undergoes HR or SSA to reconstitute functional GFP, and the cells will turn...set of plasmids required to make singly or multiply phosphorylated proteins (B40 OTS and pCRT7-GFP). To ...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available...cell free system allows for a relatively simple, controlled “one-pot” reaction capable of reproducing useful... Xiao Wang Lab has recently deposited a set of plasmids that helps identify base edited cell populations...populations without end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter...coding sequence for a green fluorescent protein (GFP). The success of the A-to-G conversion could be seen...

...sugars (including lactose), and a lac operon controlled GFP. When the enzyme breaks down the HMO of interest...lactose released activates expression of GFP from the lac operon. GFP expression can then be used to determine...Xin hypothesized that she could make enzymes controllable by tethering them to Dronpa, a fluorescent protein...their ability to be precisely spatio-temporally controlled as a key advantage over other inducible enzyme...Photosactivatable Fluorescent Proteins Use Light to Control CRISPR Genome Editing Choose the Brightest Fluorescent...Addgene.org Find Fluorescent Protein Biosensors Browse Plasmids from the Tsien Lab Browse the Michael Davidson...second sensor is a fusion protein consisting of sfGFP fused to a His-tagged ABC transporter known to bind...

...amazing plasmid technologies developed by our community of depositors. With over 40,000 plasmids avaliable...amazing new plasmid tools that have come through our doors in 2015. Do you have a favorite new plasmid tool ...temporal control over CRISPR/Cas9 mediated genome modification and gene expression. The plasmids associated...system plasmids are available through Addgene and are already very popular. Use these plasmids to increase...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins... can visualize: Cell Structure Plasmids: Cell Structure Plasmids: ER mCherry-Sec61 beta BFP-Sec61...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...

...both positively and negatively controls the output. In this case, plasmid copy number is the input and ...to decouple gene expression and plasmid number. In a suite of plasmids modified from pSC101 with copy ...of all types of plasmids, the stabilized promoters were also tested in different plasmid backbones that...stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, ...trans-acting RepA protein under the control of IPTG. Because RepA is a plasmid replication initiator protein... constitutive promoter upstream of GFP increased expression of GFP six-fold, but a stabilized promoter...genome or a plasmid. Thus, a stabilized promoter can allow a system to be moved from a plasmid to the genome...

...applications. This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB)-2A-GFP (PX458) - This...and an envelope expressing plasmid, such as our second most requested plasmid of all time, pMD2.G. Find...pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR... top requested plasmid from 2016. You may notice the “V2.0” at the end of this plasmid which indicates...prior version of this plasmid. Indeed the first version is the 15th most popular plasmid of all time, but ...envelope expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini....1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...

...via HDR using the supplied donor plasmid will express GFP. If there’s GFP expression, presumably the endogenous...  A donor plasmid that contains a puromycin resistance gene linked to an in-frame H2B-GFP. This visual...also found in the reporter gene). A control RFP expression plasmid.  With this set up, all successfully...integrated GFP reporter leaving you with just your desired genomic edit.  Find SRIRACCHA plasmids at Addgene...RFP to GFP expressing cells provides a real-time read out of Cas9 activity.  By using the RFP:GFP ratio...in the genome) and an out-of-frame H2B (histone)-GFP reporter into the genome of mammalian cells.  ...the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The Madison lab also created...

...self-assembling split GFP (GFP1-9 and GFP10-11) by flipping a single β-strand of GFP, hence the name FlipGFP. FlipGFP...apoptosis reporter, ZipGFP, which utilized the self-assembling split GFP (GFP1-10 and GFP11) to increase fluorescence... reporters (Zhang et al., 2019). How does FlipGFP work? GFP contains 11 β-strands and one central ɑ-helix...Lab used FlipGFP to track the spatiotemporal dynamics of apoptosis during embryo development. GFP fluorescence...diseases and disorders. You can find the plasmids for FlipGFP and FlipCherry at Addgene. References ...system, and cell turnover. Apoptosis is a highly controlled process that is triggered by internal and external...protein-protein interaction sensor based on tripartite split-GFP association." Scientific reports 3 (2013): 2854. ...

...Table 1). The four systems Name Function Plasmid Transduction Marker Mouse Strain C-CHIME... out a pair of genes pXPR_219 Violet-excited GFP (vex) Rosa26-Cas9 I-CHIME Inducible knockout...knockout of a single gene pXPR_068 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 L-CHIME ...types (lineage-based) pXPR_070 Violet-excited GFP (vex) Lineage-Cre; Rosa26-Cas9 S-CHIME Knocks...of genes sequentially pXPR_220 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 Table 1: X-CHIME...Concept These systems were benchmarked on positive control genes in CD4+ and CD8+ T cells. To demonstrate ...  The X-CHIME deposit also includes pXPR_071, a plasmid that leads to higher titer lentivirus preparations...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...sender” cells express GFP on the surface and “receiver” cells express an anti-GFP nanobody on the cell ...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...head over to our CRISPR Plasmids and Resources page.  Plasmids for delivering CRISPRi components by a single...inducible degron system for C. elegans New CRISPR plasmids New items from the viral service Epigenetic ... Haynes at Emory recently contributed a set of plasmids to the repository that will help epigenetic researchers...

...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ...protocol to transfect mammalian cells with your plasmid of interest. Protocols...channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM high ..., the plasmid should also be propagated in an endonuclease negative E. coli strain such as NEB stable....is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1–2 ...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...

...Fluorescent Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite multiple studies ...protein or Enhanced Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general... a tag with a serious drawback. The standard EGFP plasmid that used to be sold by Clontech and is in a...in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently...best solution is to use truly monomeric FPs. EGFP and other GFP family members (superfolder, Emerald, CFP...

...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, may be...Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning and expressing... of interest with GFP expression.  Cloning and expressing RNAi pMKO.1 puro (Plasmid #8452) RNA interference...Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral Barcoding...expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has been used by The RNAi Consortium...

...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. .... Today, it's time for another edition of Hot Plasmids!  Here's what you'll find in this post:  New...., 2023 under CC-BY 4.0 License. Find PE6 plasmids here!  Doman, J.L., Pandey, S., et al. (2023)....therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific... enzyme, Lee and Cheah et al. achieved precise control over when and where the enzyme is active, drastically...pulse-chase experiments, and more. Find LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID...LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments open up new possibilities...

...feature some recently deposited plasmids every quarter. The “hot plasmids” from last March were quite popular...behalf of scientists. 9. Plasmids 101: Positive and Negative Selection for Plasmid Cloning by Jennifer Tsang...past year: CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These...s the top 10 blog posts from the year. 10. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent... about mNeonGreen, mTFP1, and mWasabi and find plasmids containing these fluorescent proteins at Addgene... PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection by ...Joachim Goedhart shares some strategies for generating GFP fusion proteins. From considering protein size and...

..., VVDfast to control recruitment of an “anchor” to microtubules and iLID-SspB to control the recruitment...reversibly control Rho GTPase activity on a global to subcellular scale, enabling precise control over vascular...Optogenetics gives you the power to control cells or organisms with the flip of a switch. You may be ...with popular light-sensitive ion channels used to control activities like neuronal signaling — think of a... a few recently-deposited optogenetic tools to control the cytoskeleton and cell membrane dynamics. First...module, which are also available as lentiviral plasmids if you want to make a stable cell line. This design...component of the cytoskeleton – actin? Actin dynamics control cell morphology, movement, and more, often driven...

...Pitfalls When Using Fluorescent Proteins Control Protein Activity with GFP Resources on Asddgene.org Find Fluorescent...signal to noise ratio of the system, they modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly...the amount of background fluorescence from unbound GFP-dCas9. With these improvements, labeling efficiency...sequences, which recruit fluorescent proteins BFP, GFP, and RFP fused to the respective hairpin binding ...24360272. PubMed Central PMCID: PMC3918502. Find plasmids from this publication at Addgene. 2. Ma, Hanhui...25713381. PubMed Central PMCID: PMC4364232. Find the plasmids from this publication at Addgene. 3. Ma, Hanhui...27088723. PubMed Central PMCID: PMC4864854. Find the plasmids from this publication at Addgene. 4. Kiani, Samira...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        AAV delivery... Figure 1: The base editor is split into two plasmids and co-transfected into cells for reconstitution...levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved...voltage indicators, however, fall short compared to GFP-based sensors in terms of response and brightness...overcome this overlap, Yiyang Gong’s lab created two plasmids containing a red-shifted Ace-mScarlet, a fusion...

...cloning plasmids, expression plasmids, gene knock-down plasmids, reporter plasmids, viral plasmids, and ... of scientists and plasmid lovers - serves as an introduction to plasmids. Plasmids 101 will provide you...quantity inside a cell Plasmids 101: Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor...be asked to design, modify, or construct a plasmid. A plasmid is a small circular piece of DNA found in... components that make up a plasmid and why each is important. Our “Plasmids 101” series designed to educate...advancing the field.   What is a plasmid? At their most basic level, plasmids are small circular pieces of...virulence. All natural plasmids contain an origin of replication (which controls the host range and copy...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...They generated plasmids tagged with EB3 to mark the growing tips of microtubuli and plasmids tagged with ...and dCas9-VPR targeting plasmids can be found at Addgene. Check out these plasmids for piggyBac gRNA delivery...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head... Find the plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting... Find the plasmids at Addgene! A new tool for synthetic biology uses CRISPRi to control gene expression...

.... pBLADEONLY: This plasmid contains only BLADE and is based on the pTRC99a plasmid. It does not include...Their system is based on the AraC protein that controls transcription from the PBAD promoter. “AraC is...AraC and other promoter systems, head over to our Plasmids 101: Inducible Promoters article.) Modifying AraC...reporter downstream of the PBAD promoter. To use this plasmid with your gene of interest, you would replace mCherry...PBAD promoter. If you already have a pBAD33-based plasmid expressing your gene of interest, all you have ... of interest, under the pBAD promoter, can be controlled with light without the need of any cloning. ...strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter. They ...

...delivering control shRNAs, GFP, and rtTA from the Tet-on system. The first two can serve as controls in a variety...virus, and quality control data for each of the materials we distribute. Quality control info can be found...The viral preparations undergo rigorous quality control testing at Addgene meaning they come ready made...accelerate your research.   Why distribute virus? Plasmid distribution is booming, but, over the years, many...list of what’s available or suggest additional plasmids with the Viral Service Suggestion Form. Lentivirus...used to activate expression of a gene under the control of the tet operator. AAV AAVs are the workhorses...are already available allowing you to chemically control neuronal activity while studying your phenotype...

.... Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid as well as several...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...write about a recent plasmid deposit please sign up here.     Listen to the Hot Plasmids episode! Expanding...blue light.  Similarly, light can also be used to control homo-oligomerization of a protein fused to CRY2...further probe cellular function. The CRY2clust Plasmid can be found here. Park et al. Nat Commun. 2017..., the Barrick lab has deposited the pDCAF3-IS5 plasmid containing a synthetic operon that enables the ...

...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...

...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...

...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!...

..., 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_...