... in the rat APOBEC1 that decreases the number of off-target RNA edits by >390 fold and >3,800 fold in .... coli genome-wide CRISPR inhibition Covalent tethering of DNA template to Cas9 SECURE base editors Nme2Cas9...genome-wide CRISPR inhibition library Covalent tethering of DNA repair template to Cas9 Homology-directed...

... temporally control which genes are turned on or off in a given area and the can do so in a very specific...gene expression studies, which are evinced by the Tet system and visualization of single molecules via ... which allow you to easily and reversibly switch off and on your favourite signalling pathways (Figure...of the ribosome during translation, which knocks off all mRNA binding proteins as it travels along the...for Translation RNA Imaging by Coat protein Knock-off, is made up of three components: a GFP protein that...NLS-MS2-RFP) the reporter mRNA, which contains the tethering sites for the fluorescent proteins and an inducible...however, remain bound to the mRNA since they are tethered AFTER the stop codon. Thus a non-translated mRNA...

...some challenges still exist. Nucleases may cause off-target effects on regions of the genome with similar...either induce or repress Cre expression using TetON or TetOFF systems. So now hopefully creating a genetically... the cell can occur. Similar systems include tetracycline or doxycycline mediated activation of Cre to...

...designed to bind specific genes and turn them on or off, literally at the flip of a (blue) light switch. ...chromophores flavin-adenine dinucleotide and methyltetrahydrofolate, which are universally abundant. Second...apply LITEs for the transformation of neurons, she offers a tip: save yourself some time by making an AAV1...

... GFP-binding nanobodies, the project really took off. Unlike most antibodies, nanobodies, which occur ...DNA-binding domains, including the commonly used LexA and rTetR systems such that they activate gene expression ...advantages. T-DOGGs can be made drug-inducible using the rTetR system, but Cre-DOG lacks the toxicity that can ...

... on-target activity to siRNA, but with far fewer off-target effects! This characteristic probably explains... by DNMT3A or MQ1, and cytosine demethylation by Tet1. RNA Targeting: This page contains Cas13 constructs...

...receive your award, we’d love to see you show it off! To share your photo, tag @Addgene and #happydepositor... the world from the far-eastern islands,” said Tetsushi Sakuma, a lecturer at Hiroshima University who...

...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...

... 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...

...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)....

... 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...

...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...

...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...

...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...

...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...

...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...

... 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 ...

...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...

...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...

...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...

...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...

...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 ...

...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 ...

...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...

...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...

...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...

...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...

... 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...

...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...