Simultaneous monitoring of the expression of multiple genes inside of single breast carcinoma cells

被引:72
作者
Medley, CD
Drake, TJ
Tomasini, JM
Rogers, RJ
Tan, WH [1 ]
机构
[1] Univ Florida, Dept Chem, Ctr Res Bio Nano Interface, Gainesville, FL 32611 USA
[2] Univ Florida, Shands Canc Ctr, Genet Inst, Gainesville, FL 32611 USA
[3] Univ Florida, McKnight Brain Inst, Gainesville, FL 32611 USA
[4] Univ Florida, Coll Med, Dept Anesthesiol, Genet Inst, Gainesville, FL 32610 USA
关键词
D O I
10.1021/ac050881y
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Monitoring gene expression is at the center of research for a wide variety of medical, biological, and biotechnological applications. Currently no method exists for true multiple gene expression monitoring inside of a single living cell that allows for the gene expression profile of the cell to be directly compared with another single living cell. By microinjecting multiple molecular beacons with different fluorophores inside of single breast carcinoma cells and monitoring with advanced fluorescent microscopy, the expression of multiple genes can be simultaneously monitored inside single living cells. Using ratiometric analysis as a basis for the measurements allows the different gene expression levels to be compared from cell to cell. Not only does this allow differentiation of individual mRNA expression levels between multiple single cells but it also allows for mRNA expression trend analysis at the single cell level.
引用
收藏
页码:4713 / 4718
页数:6
相关论文
共 21 条
[1]   Visualizing the distribution and transport of mRNAs in living cells [J].
Bratu, DP ;
Cha, BJ ;
Mhlanga, MM ;
Kramer, FR ;
Tyagi, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (23) :13308-13313
[2]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
[3]   Methods for visualizing RNA processing and transport pathways in living cells [J].
Dirks, RW ;
Molenaar, C ;
Tanke, HJ .
HISTOCHEMISTRY AND CELL BIOLOGY, 2001, 115 (01) :3-11
[4]   Molecular beacon DNA probes and their bioanalytical applications [J].
Drake, TJ ;
Tan, WH .
APPLIED SPECTROSCOPY, 2004, 58 (09) :269A-280A
[5]   Stochastic gene expression in a single cell [J].
Elowitz, MB ;
Levine, AJ ;
Siggia, ED ;
Swain, PS .
SCIENCE, 2002, 297 (5584) :1183-1186
[6]   Designing a novel molecular beacon for surface-immobilized DNA hybridization studies [J].
Fang, XH ;
Liu, XJ ;
Schuster, S ;
Tan, WH .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1999, 121 (12) :2921-2922
[7]   Gene expression and the myth of the average cell [J].
Levsky, JM ;
Singer, RH .
TRENDS IN CELL BIOLOGY, 2003, 13 (01) :4-6
[8]   Peptide-linked molecular beacons for efficient delivery and rapid mRNA detection in living cells [J].
Nitin, N ;
Santangelo, PJ ;
Kim, G ;
Nie, SM ;
Bao, G .
NUCLEIC ACIDS RESEARCH, 2004, 32 (06) :e58
[9]   Immunodiagnosis of parasitic diseases with synthetic peptides [J].
Noya, O ;
Patarroyo, ME ;
Guzmán, F ;
de Noya, BA .
CURRENT PROTEIN & PEPTIDE SCIENCE, 2003, 4 (04) :299-308
[10]   Real-time monitoring of intracellular mRNA hybridization inside single living cells [J].
Perlette, J ;
Tan, WH .
ANALYTICAL CHEMISTRY, 2001, 73 (22) :5544-5550