Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia.
AuthorsCecic I, Chan DA, Sutphin PD, Ray P, Gambhir SS, Giaccia AJ, Graves EE
JournalMol Imaging
PubMed ID17711777
'Reporter gene techniques have been applied toward studying the physiologic phenomena associated with tumor hypoxia, a negative prognostic indicator. The purpose of this study was to assess the potential adverse effects of hypoxic conditions on the effectiveness of four commonly used reporter genes: Renilla luciferase, monomeric red fluorescent protein, thymidine ... More
In vivo imaging of beta-galactosidase activity using far red fluorescent switch.
AuthorsTung CH, Zeng Q, Shah K, Kim DE, Schellingerhout D, Weissleder R
JournalCancer Res
PubMed ID14996712
'beta-Galactosidase (beta-gal) has been widely used as a transgene reporter enzyme, and several substrates are available for its in vitro detection. The ability to image beta-gal expression in living animals would further extend the use of this reporter. Here we show that DDAOG, a conjugate of beta-galactoside and 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one) (DDAO), ... More
beta-Galactosidase activity assay using far-red-shifted fluorescent substrate DDAOG.
AuthorsGong H, Zhang B, Little G, Kovar J, Chen H, Xie W, Schutz-Geschwender A, Olive DM,
JournalAnal Biochem
PubMed ID19103143
beta-Galactosidase (beta-gal) is commonly used as a reporter gene in biological research, and a wide variety of substrates have been developed to assay its activity. One substrate, 9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) beta-d-galactopyranoside (DDAOG), can be cleaved by beta-gal to produce 7-hydroxy-9H(I,3-dichloro-9,9-dimethylacridin-2-one) (DDAO). On excitation, DDAO generates a far-red-shifted fluorescent signal. Using this substrate, ... More
Fundamental to eukaryotic cell signaling is the regulation of protein function by directed localization. Detection of these events has been largely qualitative owing to the limitations of existing technologies. Here we describe a method for quantitatively assessing protein translocation using proximity-induced enzyme complementation. The complementation assay for protein translocation (CAPT) ... More
Quantification of beta-galactosidase activity after non-viral transfection in vivo.
AuthorsZinselmeyer BH, Beggbie N, Uchegbu IF, Schätzlein AG
JournalJ Control Release
PubMed ID12932652
The limited efficacy of non-viral gene delivery systems currently hampers their wider therapeutic use. In order to further develop novel gene delivery systems, it is important to quantify their efficacy. Many reporter gene assays have limitations when being used to quantify expression in vivo. We have developed a simple assay ... More
A combinatorial network of evolutionarily conserved myelin basic protein regulatory sequences confers distinct glial-specific phenotypes.
AuthorsFarhadi HF, Lepage P, Forghani R, Friedman HC, Orfali W, Jasmin L, Miller W, Hudson TJ, Peterson AC
JournalJ Neurosci
PubMed ID14614079
Myelin basic protein (MBP) is required for normal myelin compaction and is implicated in both experimental and human demyelinating diseases. In this study, as an initial step in defining the regulatory network controlling MBP transcription, we located and characterized the function of evolutionarily conserved regulatory sequences. Long-range human-mouse sequence comparison ... More
Multiparameter flow cytometric detection and quantification of senescent cells in vitro.
Authors
JournalBiogerontology
PubMed ID32776262
Identification of a Sacral, Visceral Sensory Transcriptome in Embryonic and Adult Mice.
Authors
JournalEneuro
PubMed ID31996391
Modulation of therapy-induced senescence by reactive lipid aldehydes.
AuthorsFlor AC, Doshi AP, Kron SJ,
JournalCell Death Discov
PubMed ID27453792
Current understanding points to unrepairable chromosomal damage as the critical determinant of accelerated senescence in cancer cells treated with radiation or chemotherapy. Nonetheless, the potent senescence inducer etoposide not only targets topoisomerase II to induce DNA damage but also produces abundant free radicals, increasing cellular reactive oxygen species (ROS). Toward ... More