- Novel photocleavable universal support for oligonucleotide synthesis
-
A novel photocleavable universal support for the automated solid phase synthesis of oligonucleotides is described. The linker between the growing oligonucleotide chain and CPG support contains a nucleophilic amine protected with a photo-cleavable group. On exposure to UV light, this group is detached and the free amine affords cleavage of the oligonucleotide from the support. The use of long wavelength UV light avoids damage to the DNA.
- Anderson, Emma,Brown, Tom,Picken, Douglas
-
-
Read Online
- METHODS AND COMPOSITIONS
-
The invention relates to genetic incorporation of 2,3-diamino propionic acid (DAP) into polypeptides, to unnatural amino acids comprising DAP, to a tRNA synthetase for charging tRNA with unnatural amino acids comprising DAP, and to methods of using the resulting polypeptides, for example in capturing substrates and/or intermediates in enzymatic reactions. The invention also relates to compounds of formula (I) or (II): or salts, solvates, tautomers, isomers or mixtures thereof.
- -
-
Page/Page column 66; 70; 71
(2020/05/29)
-
- A Photoactivatable Formaldehyde Donor with Fluorescence Monitoring Reveals Threshold to Arrest Cell Migration
-
Controlled light-mediated delivery of biological analytes can enable the investigation of highly reactivity molecules within living systems. As many biological effects are concentration dependent, it is critical to determine the location, time, and quantity of analyte donation. In this work, we have developed the first photoactivatable donor for formaldehyde (FA). Our optimized photoactivatable donor, photoFAD-3, is equipped with a fluorescence readout that enables monitoring of FA release with a concomitant 139-fold fluorescence enhancement. Tuning of photostability and cellular retention enabled quantification of intracellular FA release through cell lysate calibration. Application of photoFAD-3 uncovered the concentration range necessary for arresting wound healing in live cells. This marks the first report where a photoactivatable donor for any analyte has been used to quantify intracellular release.
- Chan, Jefferson,Ibarra, Gabriela E.,Krishnamurthy, Vishnu,Pino, Nicholas W.,Smaga, Lukas P.
-
supporting information
(2020/01/31)
-
- Photo-controlled cell-specific metabolic labeling of RNA
-
Elucidating gene expression programs within a cell-specific manner is a grand challenge for biologists. Harder still is the ability to have kinetic control over such experiments. Metabolic labeling with bioorthogonally-functionalized metabolic intermediates provides a means to profile RNA expression in a cell-specific manner, but there is still a lack of kinetic resolution. Herein we present the synthesis and evaluation of photocaged metabolic uracil intermediates. We compare the photo-decaging properties and demonstrate their utility in metabolic labeling experiments in a cell-specific manner. We anticipate that our approach will have far-reaching impact as it provides control over tagging of nascent RNA.
- Feng,Li,Spitale
-
supporting information
p. 5117 - 5120
(2017/07/10)
-
- Isotope effects in photochemistry: Application to chromatic orthogonality
-
Equation Presented The main challenge in developing new wavelength-specific photolabile protecting groups is the rigorous control of the photolysis rate. This rate is controlled by two factors: the chromophore absorbance and the reaction quantum yield. Fine-tuning the properties by changing substituents or structural features is difficult, because both factors are independently affected. By the use of the kinetic isotope effect, we could tune the quantum yield without altering the absorbance, and hence control the overall reaction rate. We exemplified this approach with chromatically orthogonally protected diesters.
- Blanc, Aurelien,Bochet, Christian G.
-
p. 2649 - 2651
(2008/02/08)
-
- Protection and labelling of thymidine by a fluorescent photolabile group
-
A fluorescent photolabile group including coumarin and MeNPOC moieties was synthesized to protect 5′-OH terminal function of thymidine (T). Its photochemical and photophysical properties were studied, in particular the photocleavage (photodeprotection under a 365-nm irradiation) is only lowered by a factor of two by addition of the fluorophore. Fluorescence properties of the coumarin probe are not changed upon irradiation, which is satisfactory for the application required, i.e., in situ synthesis of DNA microarrays.
- Muller, Caroline,Even, Pascale,Viriot, Marie-Laure,Carre, Marie-Christiane
-
p. 3735 - 3741
(2007/10/03)
-
- The efficiency of light-directed synthesis of DNA arrays on glass substrates
-
New methods based on photolithography and surface fluorescence were used to determine photodeprotection rates and stepwise yields for light-directed oligonucleotide synthesis using photolabile 5'-(((α-methyl-2- nitropiperonyl)-oxy)carbonyl)(MeNPOC)-2'-deoxynucleoside phosphoramidites on planar glass substrates. Under near-UV illumination (primarily 365 nm) from a mercury light source, the rate of photoremoval of the MeNPOC protecting group was found to be independent of both the nucleotide and length of the growing oligomer (t( 1/4 ) = 12 s at 27.5 mW/cm2). A moderate dependence on solvent polarity was observed, with photolysis proceeding most rapidly in the presence of nonpolar solvents or in the absence of solvent (e.g., t( 1/4 ) = 10 - 13 s at 27.5 mW/cm2). In solution, the photolysis rate was linearly dependent on light intensity over the range 5-50 mW/cm2. Average stepwise yields for the synthesis of dodecamer oligonucleotides were in the range of 92-94%, using monomers based on N6-(phenoxyacetyl)-2'-deoxyadenosine, N2- isobutyryl-2'-deoxyguanosine, N4-isobutyryl-2'-deoxycytidine, and thymidine. By comparison, an efficiency of 98%/step was obtained using a conventional 5'-dimethoxytrityl monomer with acid deprotection on the same support. The lower yields associated with the photochemical process appears to be due to incomplete recovery of free 5'-hydroxyl groups after photolysis on the support, although high yields of 5'-OH nucleosides (≤96%) are consistently observed when 5'-MeNPOC monomers are photolyzed in solution.
- McGall, Glenn H.,Barone, Anthony D.,Diggelmann, Martin,Fodor, Stephen P. A.,Gentalen, Erik,Ngo, Nam
-
p. 5081 - 5090
(2007/10/03)
-
