...promoter driving tTA; contains insulator sequence; plasmid pEnt L1L3 rtTA-3 is a rtTA version tTA Off Hsiao ...tetracycline-controlled transactivator (tTA) was developed. tTA was created by fusing tetR with the C-terminal..., which controls GFP expression tTA Off Verma 26803 pEnt L1L3 EF1a-tTA-2 Contains Gateway L1L3 sites and...34856 fos-tTA Mouse fos promoter plus part of the first intron (-764/+918) and fos 3'UTR tTA Off Mayford...transregulation. Dox binds with high affinity to tTA and rtTA; thus, dox can be used in both tetracycline ...interest. ID Plasmid Description Co-expressed tTA, rtTA, or TetR On or Off PI 21916 Tet-pLKO-neo 3rd generation...minimal CMV and is silent in absence of binding of tTA or rtTA None On Vogelstein 25735 pSLIK-Neo Lentiviral...

...99118 pAAV-CAG-tTA CAG-driven, constitutive Expression of the tet-off transactivator (tTA) 2 Gradinaru ...-ihSyn1-tTA Inducible Synapsin promoter (ihSyn) Expression of the tet-off transactivator (tTA) with a ...feedback loop for amplified tTA expression. 1 Gradinaru 99121 pAAV-ihSyn1-DIO-tTA Cre-dependent and inducible... tet-off transactivator (tTA) with a positive feedback loop for amplified tTA expression. 1 Gradinaru ...

...non-standard), expressing Rep2 and PHP.eB Cap controlled by tTA-TRE amplification system Gradinaru 103006 pUCmini-iCAP-PHP.S...non-standard), expressing Rep2 and PHP.S Cap controlled by tTA-TRE amplification system Gradinaru 127847 pUCmini-iCAP-PHP.V1...non-standard), expressing Rep2 and PHP.V1 Cap controlled by tTA-TRE amplification system Gradinaru 175004 pUCmini-iCAP-AAV.CAP-B10...non-standard), expressing Rep2 and B10 Cap controlled by tTA-TRE amplification system Gradinaru 175005 pUCmini-iCAP-AAV.CAP-B22...non-standard), expressing Rep2 and B22 Cap controlled by tTA-TRE amplification system Gradinaru 185136 pUCmini-iCAP-AAV.MaCPNS1...), expressing Rep2 and MaCPNS1 Cap controlled by tTA-TRE amplification system Gradinaru 185137 pUCmini-iCAP-AAV.MaCPNS2...), expressing Rep2 and MaCPNS2 Cap controlled by tTA-TRE amplification system Gradinaru RepCap Plasmids...

...levels of XFPs - Reduce the amount of tTA. Paradoxically, too much tTA leads to a reduced expression level...transcription. It is critical to express a minimal amount of tTA to achieve highest expression levels. Clearing large...

...March.  Addgene is now offering an AAV2 CAG-driven tTA expresser from the Gradinaru lab. This is useful ...AAV8 Optogenetics Boyden New serotypes pAAV-CAG-tTA AAV2 Molecular Tools Gradinaru New viral service ...

...pPRIME-Tet-GFP-FF3) and one with either tTA or rtTA (FUW-M2rtTA). In this kind of approach, a population...contrast, in the Tet-on system, tTA has been modified (and renamed rtTA) to bind TRE only in presence of...vector systems containing both your transgene and tTA/rtTA (pINDUCER20, pINDUCER21) have been developed, ...Doxycycline (Dox), the Tet-controlled transactivator (tTA, also expressed by your transfer vector) binds to...population of cells might express only the tTA or your transgene resulting in low inducibility and potentially...

... Name 99120  AAV1 pAAV-ihSyn1-tTA 99121  AAV1 pAAV-ihSyn1-DIO-tTA Viral vectors coming soon! These...

...created the tetracycline-controlled transactivator (tTA). They fused TetR with the transcriptional activation...domain VP16 taken from the herpes simplex virus. tTA binds to promoters containing TetO elements (often...tetracycline or one of its derivatives is added, it binds tTA, resulting in a confirmation change that prevents...

...Gordon Fishell 99120-AAV1 pAAV-ihSyn1-tTA An AAV genome that expresses tTA from the hSyn1 promoter that contains...amplifed expression of the tTA Viviana Gradinaru 99121-AAV1 pAAV-ihSyn1-DIO-tTA An AAV genome with Cre-dependent...-dependent expression of the tTA from the hSyn1 promoter. Contains a positive feedback loop for amplifed...amplifed expression of the tTA Viviana Gradinaru 99130-AAV1 pAAV-mDlx-NLS-mRuby2 An AAV genome that expresses...Gradinaru 100798-AAV1 pAAV-syn-FLEX-splitTVA-EGFP-tTA helper virus for monosynaptic tracing; to be coinjected...to be coinjected with pAAV-syn-FLEX-splitTVA-EGFP-tTA Ian Wickersham 104052-AAVPHP.V1 pAAV-CAG-DIO-EYFP...

...control of an inducer, i.e. tet-off transactivator (tTA), is injected at a high dose to maximize the number...high (figure 4e) and low doses (figure 4f) of the tTA inducer virus, but the desired sparsity of labeling...

...AAV1 Wickersham 100798 pAAV-syn-FLEX-splitTVA-EGFP-tTA These AAV together can be used to complement deletion-mutant...

....), and tetracycline drug induction systems (ttA or rtTA) are between 1 to 2 kb in length. There are plenty...

...with tTALE to “stain” DNA. TALE proteins can bind specific DNA sequences which brings the attached fluorescent...They created plasmids for purification of tTALE-eGFP and tTALE-mCherry. In comparison to intercalating ...Tsang Figure 4: Double stranded DNA bound by tTALE-FP with the fluorescent protein fluorescing green...high salt conditions - up to 100 mM NaCl. These tTALE-FPs also have higher DNA binding affinity for AT-rich...stained evenly in 1x TE buffer. The lab also used tTALE-FP in a single molecule-fluorescence assay to monitor...molecules in real-time. Find plasmids expressing tTALE-FP Shin et al., Sci. Rep. 2019. https://doi.org/...

...with these new photoconvertible proteins. With Sattarzadeh et al.’s discovery that GFP itself is photoconvertible...GFP-labeled plant cells at 405 nm instead of 488 nm, Sattarzadeh et al. noticed that GFP had turned red. They ... cytosol via cytoplasmic strands. Image from Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications ... photoconversion signal may not be observed. Sattarzadeh et al. found that only the protonated form of...appropriate for high-pH cellular compartments. Sattarzadeh et al. have shown that GFP can be rapidly and...empty backbones containing GFP variants used by Sattarzadeh et al., as well as a wide variety of other fluorescent...refresher, see our FP blog posts.     References Sattarzadeh A, Saberianfar R, Zipfel WR, Menassa R, Hanson...

...bolster and modulate our immune defenses to better attack cancer. Combing our knowledge with our bodies’ ...concept of immunoediting, by way of favorably attaining the equilibrium stage. This can be called “stalemate...created by the cancer cells to shield themselves from attack. It is a region of incessant chemical activity,...that suppress APCs. T cells that would normally attack the cancer cells are also suppressed by chemical...microenvironment TGFβ, IL-10, can also help the immune system attack tumors. Attempts have been made to use whole cell...collaborative therapies must be explored. When attacking cancer, multiple immunotherapy targets can be ...

...endogenous molecular target, meaning the dye will be attached to the target through binding of the molecular...encoded tag is required. These dyes can also be attached to an antibody to detect various targets, such...or glycans. Fluorophores can also be covalently attached to self-labeling proteins. Some common examples...covalently reacts with a small substrate ligand motif attached to a fluorophore. When paired with a protein of...higher specificity, increased versatility in probe attachment, and compatibility with far-red and near-infrared...Isothiocyanate-conjugated Antibody. Salari, M., Bitounis, D., Bhattacharya, K., Pyrgiotakis, G., Zhang, Z., Purington, ...

...cofactor that in its active form is covalently attached to the active site of key metabolic enzymes including...Southern/Northern blotting Cell surface labelling Attaching polypeptides to a substrate Flow cytometry/fluorescence-activated...Photobiotin acetate consists of a biotin molecule attached to a photoreactive arylazide group. Under strong...methods. Scientists have commonly used this method to attach proteins or polypeptides to a solid substrate (... & Cronan, J. E. “Molecular Biology of Biotin Attachment to Proteins; Symposium: Nutrition, Biochemistry...

...is no coming back. The whole lab is emotionally attached to this disease.” He says that the award from ...Patients affected by AHC suffer from recurrent attacks of unilateral paralysis (hemiplegia) that are often...often painful and accompanied by migraines. Those attacks can be brought on by a number of stressors, including...syndrome (WS4) by Pediatrician Duangrurdee Wattanasirichaigoon carried different mutations in a gene called...

...irreversibly with a small substrate ligand motif attached to a fluorophore (Figure 1b). Engineered ligases...phosphopantetheinyltransferase) – These enzymes catalyze the covalent attachment of a fluorophore ligand to a peptide tag (Figure...bioorthogonal chemical reactions (i.e., click chemistry) to attach a fluorophore at the specific site of incorporation... the speed and selectivity of the fluorophore attachment, the brightness of the resulting conjugate, and...

...specifically at ‘TTAA’ sites randomly dispersed in the genome. The predicted frequency of ‘TTAA’ in the genome...sequence from the transposon vector to one of many TTAA sequences distributed throughout the genome. We ...donor-transposon (carrying your cargo of interest) into random ‘TTAA’ sites throughout the genome. When using piggyBac...