TY - JOUR
T1 - Cellular arrays for large-scale analysis of transcription factor activity
AU - Bellis, Abigail D.
AU - Peňalver-Bernabé, Beatriz
AU - Weiss, Michael S.
AU - Yarrington, Michael E.
AU - Barbolina, Maria V.
AU - Pannier, Angela K.
AU - Jeruss, Jacqueline S.
AU - Broadbelt, Linda J.
AU - Shea, Lonnie D.
PY - 2011/2
Y1 - 2011/2
N2 - Identifying molecular mechanisms or therapeutic targets is typically based on large-scale cellular analysis that measures the abundance of mRNA or protein; however, abundance does not necessarily correlate with activity. We report a method for direct large-scale quantification of active pathways that employs a cellular array with parallel gene delivery of constructs that report pathway activity. The reporter constructs encode luciferase, whose expression is influenced by binding of transcription factors (TFs), which are the downstream targets of signaling pathways. Luciferase levels are quantified by bioluminescence imaging (BLI), which allows for rapid, non-invasive measurements. Activity profiles by BLI of 32 TFs were robust, consistent, and reproducible, and correlated with standard cell lysis techniques. The array identified five TFs with differential activity during phorbol-12-myristate-13-acetate (PMA)-induced differentiation of breast cancer cells. A system for rapid, large-scale, BLI quantification of pathway activity provides an enabling technology for mechanistic studies of cellular responses and processes.
AB - Identifying molecular mechanisms or therapeutic targets is typically based on large-scale cellular analysis that measures the abundance of mRNA or protein; however, abundance does not necessarily correlate with activity. We report a method for direct large-scale quantification of active pathways that employs a cellular array with parallel gene delivery of constructs that report pathway activity. The reporter constructs encode luciferase, whose expression is influenced by binding of transcription factors (TFs), which are the downstream targets of signaling pathways. Luciferase levels are quantified by bioluminescence imaging (BLI), which allows for rapid, non-invasive measurements. Activity profiles by BLI of 32 TFs were robust, consistent, and reproducible, and correlated with standard cell lysis techniques. The array identified five TFs with differential activity during phorbol-12-myristate-13-acetate (PMA)-induced differentiation of breast cancer cells. A system for rapid, large-scale, BLI quantification of pathway activity provides an enabling technology for mechanistic studies of cellular responses and processes.
KW - Bioluminescence imaging
KW - Cellular array
KW - Transcription factor activity
UR - http://www.scopus.com/inward/record.url?scp=78650224439&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78650224439&partnerID=8YFLogxK
U2 - 10.1002/bit.22916
DO - 10.1002/bit.22916
M3 - Article
C2 - 20812256
AN - SCOPUS:78650224439
SN - 0006-3592
VL - 108
SP - 395
EP - 403
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
IS - 2
ER -