Single-cell analysis of siRNA-mediated gene silencing using multiparameter flow cytometry

Cytometry Part A

Volumn 69, Issue 2, 2006, Pages 59-65

  Chan, Steven M ^a   Olson, Janelle A ^a   Utz, Paul J ^a,b  

^a STANFORD UNIVERSITY SCHOOL OF MEDICINE   (United States)

^b STANFORD UNIVERSITY   (United States)

Author keywords

Flow cytometry; Jurkat; Lck; Multiparameter; RNA interference; Signal transduction; siRNA; T lymphocytes

Indexed keywords

CD3 ANTIGEN; CD4 ANTIGEN; CD69 ANTIGEN; CD8 ANTIGEN; MITOGEN ACTIVATED PROTEIN KINASE P38; PHYTOHEMAGGLUTININ; PROTEIN KINASE LCK; PROTEIN KINASE ZAP 70; SMALL INTERFERING RNA; T LYMPHOCYTE RECEPTOR; LEUKOCYTE ANTIGEN; LYMPHOCYTE ANTIGEN RECEPTOR; T LYMPHOCYTE ANTIGEN; ZAP70 PROTEIN, HUMAN;

ARTICLE; CONTROLLED STUDY; ELECTROPORATION; ENZYME PHOSPHORYLATION; FLOW CYTOMETRY; GENE SILENCING; HUMAN; HUMAN CELL; IMMUNOFLUORESCENCE MICROSCOPY; LEUKEMIA CELL LINE; PERIPHERAL BLOOD MONONUCLEAR CELL; PRIORITY JOURNAL; PROTEIN EXPRESSION; QUANTITATIVE ANALYSIS; RNA INTERFERENCE; SEQUENCE HOMOLOGY; SIGNAL TRANSDUCTION; T LYMPHOCYTE; WESTERN BLOTTING; CELL LINE; CHEMISTRY; GENE EXPRESSION REGULATION; GENETIC TRANSFECTION; GENETICS; METHODOLOGY; PHYSIOLOGY; TUMOR CELL LINE; UPREGULATION;

ANTIGENS, CD; ANTIGENS, CD4; ANTIGENS, CD8; ANTIGENS, DIFFERENTIATION, T-LYMPHOCYTE; CELL LINE; CELL LINE, TUMOR; FLOW CYTOMETRY; GENE EXPRESSION REGULATION; GENE SILENCING; HUMANS; JURKAT CELLS; LYMPHOCYTE SPECIFIC PROTEIN TYROSINE KINASE P56(LCK); RECEPTORS, ANTIGEN, T-CELL; RNA, SMALL INTERFERING; SIGNAL TRANSDUCTION; T-LYMPHOCYTES; TRANSFECTION; UP-REGULATION; ZAP-70 PROTEIN-TYROSINE KINASE;

EID: 33645055687     PISSN: 15524922     EISSN: 15524930     Source Type: Journal    
DOI: 10.1002/cyto.a.20209     Document Type: Article

References (26)

1. Dykxhoorn DM, Novina CD, Sharp PA. Killing the messenger: short RNAs that silence gene expression. Nat Rev Mol Cell Biol 2003;4:457-467.
2. Wu W, Hodges E, Redelius J, Hoog C. A novel approach for evaluating the efficiency of SiRNAs on protein levels in cultured cells. Nucleic Acids Res 2004;32:e17.
3. Frisa PS, Lanford RE, Jacobberger JW. Molecular quantification of cell cycle-related gene expression at the protein level. Cytometry 2000;39:79-89.
4. McManus MT, Haines BB, Dillon CP, Whitehurst CE, van Parijs L, Chen J, Sharp PA. Small interfering RNA-mediated gene silencing in T lymphocytes. J Immunol 2002;169:5754-5760.
5. Perez OD, Nolan GP. Simultaneous measurement of multiple active kinase states using polychromatic flow cytometry. Nat Biotechnol 2002;20:155-162.
6. Nagy P, Arndt-Jovin DJ, Jovin TM. Small interfering RNAs suppress the expression of endogenous and GFP-fused epidermal growth factor receptor (erbB1) and induce apoptosis in erbB1-overexpressing cells. Exp Cell Res 2003;285:39-49.
7. Novina CD, Murray MF, Dykxhoorn DM, Beresford PJ, Riess J, Lee SK, Collman RG, Lieberman J, Shankar P, Sharp PA. siRNA-directed inhibition of HIV-1 infection. Nat Med 2002;8:681-686.
8. Ohba H, Zhelev Z, Bakalova R, Ewis A, Omori T, Ishikawa M, Shinohara Y, Baba Y. Inhibition of bcr-abl and/or c-abl gene expression by small interfering, double-stranded RNAs: cross-talk with cell proliferation factors and other oncogenes. Cancer 2004;101:1390-1403.
9. Palacios EH, Weiss A. Function of the Src-family kinases, Lck and Fyn, in T-cell development and activation. Oncogene. 2004;23:7990-8000.
10. Overton WR. Modified histogram subtraction technique for analysis of flow cytometry data. Cytometry. 1988;9:619-626.
11. Roederer M, Treister A, Moore W, Herzenberg LA. Probability binning comparison: a metric for quantitating univariate distribution differences. Cytometry 2001;45:37-46.
12. Chan SM, Ermann J, Su L, Fathman CG, Utz PJ. Protein microarrays for multiplex analysis of signal transduction pathways. Nat Med 2004;10:1390-1396.
13. Donaldson JG. Immunofluoresence staining. In: Morgan K, editor. Current Protocols in Cell Biology. New York: Wiley; 1998. p 4.3.1-4.3.6.
14. Mardiney M, III, Brown MR, Fleisher TA. Measurement of T-cell CD69 expression: a rapid and efficient means to assess mitogen- or antigen-induced proliferative capacity in normals. Cytometry 1996;26:305-310.
15. Cantrell D. T cell antigen receptor signal transduction pathways. Annu Rev Immunol. 1996;14:259-274.
16. Zamoyska R, Basson A, Filby A, Legname G, Lovatt M, Seddon B. The influence of the src-family kinases, Lck and Fyn, on T cell differentiation, survival and activation. Immunol Rev 2003;191:107-118.
17. Krutzik PO, Irish JM, Nolan GP, Perez OD. Analysis of protein phosphorylation and cellular signaling events by flow cytometry: techniques and clinical applications. Clin Immunol 2004;110:206-221.
18. Irish JM, Hovland R, Krutzik PO, Perez OD, Bruserud O, Gjertsen BT, Nolan GP. Single cell profiling of potentiated phospho-protein networks in cancer cells. Cell 2004;118:217-228.
19. Chow S, Patel H, Hedley DW. Measurement of MAP kinase activation by flow cytometry using phospho-specific antibodies to MEK and ERK: potential for pharmacodynamic monitoring of signal transduction inhibitors. Cytometry 2001;46:72-78.
20. Juan G, Traganos F, James WM, Ray JM, Roberge M, Sauve DM, Anderson H, Darzynkiewicz Z. Histone H3 phosphorylation and expression of cyclins A and B1 measured in individual cells during their progression through G2 and mitosis. Cytometry 1998;32:71-77.
21. Juan G, Gruenwald S, Darzynkiewicz Z. Phosphorylation of retinoblastoma susceptibility gene protein assayed in individual lymphocytes during their mitogenic stimulation. Exp Cell Res 1998;239:104-110.
22. Salvador JM, Mittelstadt PR, Guszczynski T, Copeland TD, Yamaguchi H, Appella E, Fornace AJ, Jr., Ashwell JD. Alternative p38 activation pathway mediated by T cell receptor-proximal tyrosine kinases. Nat Immunol 2005;6:390-395.
23. Veillette A, Bookman MA, Horak EM, Bolen JB. The CD4 and CD8 T cell surface antigens are associated with the internal membrane tyrosine-protein kinase p56lck. Cell 1988;55:301-308.
24. Barber EK, Dasgupta JD, Schlossman SF, Trevillyan JM, Rudd CE. The CD4 and CD8 antigens are coupled to a protein-tyrosine kinase (p561ck) that phosphorylates the CD3 complex. Proc Natl Acad Sci USA 1989;86:3277-3281.
25. Pelchen-Matthews A, Parsons IJ, Marsh M. Phorbol ester-induced downregulation of CD4 is a multistep process involving dissociation from p561ck, increased association with clathrin-coated pits, and altered endosomal sorting. J Exp Med 1993;178:1209-1222.
26. Sleckman BP, Shin J, Igras VE, Collins TL, Strominger JL, Burakoff SJ. Disruption of the CD4-p561ck complex is required for rapid internalization of CD4. Proc Natl Acad Sci USA 1992;89:7566-7570.