We narrowed to 81 results for: tet off
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TypeBlog Post...detection methods lack spatiotemporal control or an “on/off switch.” This makes them great tools for providing...speed of labeling. BioID requires 6-24 hours for tetracycline induced expression of the BirA* fusion protein...Split-BioID, Schopp et al allowed 24 hours for tetracycline induced expression and biotinylation of interacting...
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Hot Plasmids and Viral Preps - May 2021
TypeBlog Post... underwent affinity screening to select for slow off-rate binders. The best candidates were validated ...where gRNA expression is induced by anhydrous tetracycline. Three new anti-CRISPRs were identified that...Janelia Farm GENIE project have arrived! We now offer several versions of GCaMP8 vectors in serotypes ...-mRuby from Liqun Luo’s lab expresses membrane tethered GFP to enhance axonal labeling and synaptophysin-tagged... -
CRISPR Plasmids - Empty gRNA Vectors
TypeCollection... Wheeldon FgH1tUTG 70183 Mammalian/Lentiviral H1-Tet none S. pyogenes Herold lentiGuide-Crimson 70683 ...Zhang pRS416-dCas9-Mxi1 + TetR + pRPR1(TetO)-NotI-gRNA 73796 Yeast pRPR1(TetO) yes, interfere S. pyogenes... 65006 Bacteria BsaI yes, interfere S. pyogenes Koffas BPK764 65767 Bacteria BsaI yes, cut S. pyogenes...yes, nick S. pyogenes Duchek pAC5-dual-dCas9VP48-sgTetO 48237 Mammalian BbsI yes, activate S. pyogenes ...Joung AAV:ITR-U6-sgRNA (Backbone) PCB-FlPO-WPRE-syntetisk pA-UTR 68347 Mammalian/AAV none S. pyogenes Elverlov-Jakobsen...62315 Yeast none S. pyogenes URA3 Lim, Qi pRPR1(1xTetO)_gRNA_handle_RPR1t 62966 Yeast HindIII none S. ... Mammalian/Lentiviral hU6 none S. pyogenes Puro Moffat lenti sgRNA(MS2)_puro backbone 73795 Mammalian/... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog Post...learning and writing about science whenever he's not off the grid. References Bischof J, Duffraisse M, Furger...interactions that uses split fluorescent proteins tethered to potential protein interaction partners. Unlike... form the complete FP because the PIPs they’re tethered to bring them into close proximity (Figure 1).... -
Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping
TypeBlog Post... introduction of MerMAIDs, neurons can be turned off quickly and with high precision - a valuable optogenetics...latter experiments, such as transcription factors tethered to dCas9, require long time scales with constitutively... -
Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins
TypeBlog Post...another, or their fluorescence can be switched on and off reversibly. This ability to modulate fluorescence...PA-FPs come in 2 flavours- monomeric and tetrameric. Tetrameric PA-FPS are better for whole cell imaging...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate...UV-Violet Cyan/ Green >2,000 Medium 490/ 511 Kaede Tetramer UV-Violet Green/ Red ~2,000 High 572/ 580 tdEos...UV-Violet Green/ Red N/A High 573/ 584 KikGr Tetramer UV-Violet Green/ Red >2,000 High 583/ 593 mKikGR...Monomer Blue Dark/ Red 20 Low 572/ 610 FP595 Tetramer Green Dark/ Red 70 - 1000 Medium 590/ 600 ...non-invasive labelling, but in contrast, PA-FPs offer a reduced potential for photobleaching and phototoxicity... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog Post...generate a knockout or edit. This can help reduce off-target activity. Deleting a large region of the genome...by L-arabinose and anhydrotetracycline (aTet), respectively. pMAZ-SK contains an aTet-inducible gRNA and...coli multiplexing plasmid is available from the Koffas lab. Scaling up your multiplexed gRNAs with Golden... on a separate plasmid. Multiplexing in E. coli Koffas Lab CRISPathBrick Multiplex Plasmid: This system...self-destruction" after induction with L-rhamnose and aTet. CRMAGE is much more efficient than traditional ...plasmids from this paper at Addgene. 3. Sakuma, Tetsushi, et al. “Multiplex genome engineering in human... -
Special Delivery: Fluorophore Targeting for FRET Studies
TypeBlog Post...binding partners does it have and how do they come off and on in a cellular environment? If these are pressing... nonfluorescent until they bind a 6-residue tetracysteine tag (sequence: CCPGCC) that can be inserted ...interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition, since these tags are so small...et al. "Hairpin structure of a biarsenical− tetracysteine motif determined by NMR spectroscopy." Journal... -
27 Hot Plasmids from 2016
TypeBlog Post...functional knock-out via frameshift and minimize off-target effects. The library was cloned into the Liu...inserting OsTIR1 into the "safe harbor" AAVS1 locus (a tet-inducible OsTIR1 plasmid is also available). Second...actin 5C promoter. Addgene’s fly library web page offers more details and protocols, and the Liu lab’s OXfCRISPR...unique tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many of which ...context-dependent human fitness genes, the laboratory of Jason Moffat at the University of Toronto has created the Toronto...demonstrate the effectiveness of their library design, Moffat laboratory members conducted multiple screens in...hits. This work has been published in Cell. The Moffat laboratory maintains a website for the TKO library... -
Genetic Code Expansion
TypeCollection...are several things to consider. If you are working off of a previously established protocol, make sure to...Ryan Mehl 85496 pDule-Tet2.0 Tetrazine2.0 tRNA synthetase M. jannaschii Tetrazine 2.0 Bacterial TAG Ryan...Ryan Mehl 85497 pDule2-Tet2.0 Tetrazine2.0 tRNA synthetas M. jannaschii Tetrazine 2.0 Bacterial TAG Ryan...Bacterial TAG Ryan Mehl 164580 pUltraI-Tet3.0[TAA] Tet3.0RS M. barkeri Tet3.0 Bacterial TAA Ryan Mehl 172482 ...TAG Ryan Mehl 174080 pDule-Tet3.0 Tet3.0 tRNA synthetase M. barkeri Tetrazine 3.0 Bacterial TAG Ryan Mehl...NES-Flag-R2-84-MbRS Tetrazine3.0 NES-Flag-R2-84 tRNA synthetase M. barkeri Tetrazine 3.0 Mammalian Ryan...aminoacyl-tRNA synthetase M. jannaschii 1,2,4,5-tetrazine ncAAs Bacterial TAG Ryan Mehl 217361 pIDTSmart-MbPylRS... -
Custom CRISPR Screens & the Green Listed Software
TypeBlog Post...Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9." Nature biotechnology...CMM), Department of Medicine, Solna, Karolinska Institutet, in Sweden. It can be very time consuming to ...CMM), Department of Medicine, Solna, Karolinska Institutet, in Sweden. He has a particular interest in using... -
CRISPR Guide
TypeGuide...increase specificity, and decrease off-target effects Sniper-Cas9 - less off-target activity; compatible with...cleaving only one strand of target dsDNA. Off-target effects or off-target activity Cas9 cleavage at undesired...of partial homology throughout the genome, called off-targets, that can impact your experiment. There are...DSB within the target DNA, it’s unlikely that two off-target nicks will be generated close enough to cause... mutating specific amino acid residues to reduce off-target editing. Some mutations disrupt interactions...method, increased fidelity enzymes generate less off-target editing than wild type Cas9. Examples of increased...proofreading and discrimination evoCas9 - decrease off-target effects xCas9 3.7 - mutations in multiple ... -
Fluorescent Protein Guide: Biosensors
TypeCollection... tension sensor modules Grashoff Lab Tension Sensor Plasmids Carsten Grashoff Tension tCRMod tension sensors...the plasmids associated with the article. We also offer ready-to-use AAV preparations of select plasmids...sensors, ER-targeted CEPIA1er ER-mitochondria tethering by PDZD8 regulates Ca(2+) dynamics in mammalian..., reagentless biosensor for ADP based on tetramethylrhodamine-labeled ParM A fluorescent, reagentless ...reagentless biosensor for ADP based on tetramethylrhodamine-labeled ParM. ACS Chem Biol. 2010 Apr 16;5(4):415-25...single cells. Angew Chem Int Ed Engl. 2018 Jun 27. Tetsuya Kitaguchi ATP Variants of ATP sensor iATPSnFR1.0...dual-color imaging. PLoS One. 2014 Jun 24;9(6):e100252. Tetsuya Kitaguchi cAMP (cyclic AMP) Red fluorescent protein-based... -
MXS Chaining
TypeBlog Post... fluorescent protein were fused in-frame to a tethering partner, which directed localization of the fluorophore...Construct Fluorophore Excitation/emission maxima Tethering partner Subcellular localization 1 TagBFP 399nm...suited for particular downstream applications. The tradeoffs between various cloning methods are listed in ... -
Hot Plasmids: Fall 2024
TypeBlog Post...on an integrative plasmid containing tetracycline repressor TetR and resistance selection marker KanR....early embryogenesis (Figure 3). mScarlet-I3 also offered clearer signals due to reduced background signal... -
New and Upcoming Viral Vectors - June 2019
TypeBlog Post...excited to offer these useful tools to the community. Among other new biosensor tools, we now offer the jGCaMP7c...To help make this easier for scientists, we now offer trial size 20 µL samples of control AAV that scientists...control of specific neuronal populations. We now offer a Flp-expressing AAV, as well as a Cre-dependent... AAV5 ppAAV-CKIIa-stGtACR1-FusionRed New tetracycline transactivators Plasmid Serotype Name 99120... -
Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog Post...organization’s technology transfer office or other authorized office directly to make sure the deposit ...nourseothricin (clonNat), spectinomycin, streptomycin, tetracycline, combinations of the above, or other. High...to Addgene and contact your technology transfer office about the MTA. Pro Tip! Download a new spreadsheet... -
Twenty Years of Sharing: Expanding Accessibility
TypeBlog Post...distributes to per year, 2005-2023. We identified and offered a number of solutions that increased accessibility...each courier we contracted with so we can always offer the fastest shipping. However, there were issues...specific to that country. For example, when we began offering temperature-sensitive materials, we noticed a ...Culture Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial ...educational resources for each type of resource we offer. In 2013, we launched the Addgene blog for topics... -
Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering
TypeBlog Post...CRISPR/Cas9 compared to ZFNs and TALENs,” said Tetsushi Sakuma of Hiroshima University. “However, there... in their recent study, the construction system offers ease and efficiency for all-in-one CRISPR/Cas9 ... -
How to Negotiate a Successful Lab Start Up Budget
TypeBlog Post...institutes that offer shared equipment that you can use at a lower cost. Don't tether yourself to a lab...on unnecessary materials, or even a loss of the offer all together. The problem is that the discussions...turned down a faculty position because none of the offering institutes were able to provide the resources ... -
Mycoplasma Contamination: Where Does It Come From and How to Prevent It
TypeBlog Post...for up to 60% of the cell culture contamination (Uphoff, 2002). Mycoplasmas are considered to be one of...for at least two or three days before sampling (Uphoff and Drexler, 2011). Culturing on agar plates/broth...fluoroquinolone ciprofloxacin, ciprobay, zagam, baytril, tetracycline, etc. are available for mycoplasma removal from... of mycoplasma species. J Microbiol .44: 42–49 Uphoff Cc, Drexler Hg (2002) Comparative Antibiotic Eradication...10.1290/1071-2690(2002)038<0086:caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma Contamination... -
Deisseroth INTRSECT Collection
TypeCollection... 55651 pAAV-hSyn Con/Foff EYFP Cre AND NOT Flp F3/F5 No 137162 pAAV-Ef1a-Con/Foff 2.0-EYFP Cre AND NOT.../F5 Yes 55652 pAAV-hSyn Coff/Fon EYFP Flp AND NOT Cre No 231927 pAAV-Ef1a-Coff/Fon-EYFP Flp AND NOT Cre...137130 pAAV-Ef1a-Con/Foff 2.0-BFP Cre AND NOT Flp FRT/F5 Yes 137131 pAAV-Ef1a-Coff/Fon-BFP Flp AND NOT ...137133 pAAV-Ef1a-Con/Foff 2.0-mCherry Cre AND NOT Flp FRT/F5 Yes 137134 pAAV-Ef1a-Coff/Fon-mCherry Flp AND...137137 pAAV-Ef1a-Con/Foff 2.0-oScarlet Cre AND NOT Flp FRT/F5 Yes 137138 pAAV-Ef1a-Coff/Fon-oScarlet Flp ...137120 pAAV-Ef1a-Con/Foff 2.0-GCaMP6M Cre AND NOT Flp FRT/F5 Yes 137121 pAAV-Ef1a-Coff/Fon-GCaMP6M Flp AND...137123 pAAV-Ef1a-Con/Foff 2.0-GCaMP6F Cre AND NOT Flp FRT/F5 Yes 137124 pAAV-Ef1a-Coff/Fon-GCaMP6F Flp AND... -
Building Global Connections with the International Mentorship Program USA-EUROPE
TypeBlog Post...professional development and networking resources offered by programs like IMP USA-Europe not only provide...University, the Salk Institute, and the Karolinska Institutet, among others). Mentors and mentees held scheduled...for the Spanish scientific community overseas. Official representatives from different Spanish universities... -
Neurodegeneration Plasmid Collection
TypeCollection...188572 Tet-off TauRD (P301L/V337M) MAPT Myc TRE Parkinson's, FTD Franz-Ulrich Hartl 188573 Tet-off Full ...Ataxia Richard Youle 176485 Piggybac-TetO-hNurr1-blast NR4A2 TetO Parkinson's Marius Wernig 176852 UBQLN2...Ronald Hart 190811 pTet-O-APOE-E2-T2A-Puro APOE UbC Alzheimer's Ronald Hart 190812 pTet-O-APOE-E3-T2A-Puro... CMV Ataxia telangiectasia Stephen Elledge 43918 tetO-ALN NR4A2 His, V5 TRE Parkinson's John Gearhart ...T2A-Puro APOE UbC Alzheimer's Ronald Hart 190813 pTet-O-APOE-E4-T2A-Puro APOE UbC Alzheimer's Ronald Hart ... -
Transferable Skills Guide: External Collaborations
TypeBlog Post...transferable skills while working in the lab and by offering advice on areas where you might need some help...collaborations. During my graduate studies at Karolinska Institutet I started working with other groups. First within... -
Plasmids 101: Dimers and Multimers
TypeBlog Post...multimeric forms (e.g. concatemers, dimers, trimers, tetramers). Since then, multimerization has been observed...-340-4_2. PMID: 19763938. Maucksch C, Bohla A, Hoffmann F, Schleef M, Aneja MK, Elfinger M, Hartl D, Rudolph... -
Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
TypeBlog Post... secondary structure. Today, multiple companies offer long single-stranded DNA production services or ...mCherry etc.), recombinases (Cre, Flp etc.), and tetracycline drug induction systems (ttA or rtTA) are between... -
Using AAV for Neuronal Tracing
TypeBlog Post...uptake or intracellular transport machinery. Addgene offers viral preparations of AAV2-retro vectors with a...retrograde transsynaptic transfer of a protein, tetanus toxin, subsequent to its retrograde axonal transport... -
Luciferase Plasmid Collection
TypeCollection...enchancer regions in Drosopholia Herman Wijnen 126033 pTETRIS-cargo Firefly For use as a reporter of the ability...to find plasmids expressing luciferase. We also offer ready-to-use AAV preparations of select luciferase... -
Lentiviral Vector Guide
TypeGuide...to the growth medium of the host cells, they kill off any cells that have not incorporated the lentiviral...LEDGF (also known as PSIP1 or p75 ), a lentiviral tethering gene that helps facilitate integration. Despite... of experiments being performed. Your biosafety office can provide more information on your institution's...PMID: 32502836 Johnson, N. M., Alvarado, A. F., Moffatt, T. N., Edavettal, J. M., Swaminathan, T. A., &...30518911 Wells, D. W., Guo, S., Shao, W., Bale, M. J., Coffin, J. M., Hughes, S. H., & Wu, X. (2020). An analytical... -
Molecular Biology Reference
TypeGuide...reporter gene (for example, luciferase or GFP) that offers a read-out of the activity of the genetic element...high efficiency. Addgene's expanding viral service offers select ready-made AAV and lentiviral particles....England Biolabs F' proA+B+ lacIq ∆(lacZ)M15 zzf::Tn10 (TetR) ∆(ara-leu) 7697 araD139 fhuA ∆lacX74 galK16 galE15.../mL 50 µg/mL Spectinomycin 50 mg/mL 50 µg/mL Tetracycline 10 mg/mL 10 µg/mL *Note: Carbenicillin can be...