Enzymatic activity in single cells

Josephine Geertsen Keller; Magnus Stougaard; Birgitta R. Knudsen

doi:10.1016/bs.mie.2019.07.003

Enzymatic activity in single cells

Josephine Geertsen Keller, Magnus Stougaard, Birgitta R. Knudsen^*

^*Corresponding author for this work

Department of Clinical Medicine - The Department of Pathology

Research output: Contribution to book/anthology/report/proceeding › Book chapter › Research › peer-review

2 Citations (Scopus)

Abstract

With the increasing recognition of the importance in addressing cell-to-cell variations for the understanding of complex biological systems, single cell analyses are becoming increasingly important. Presented in this chapter is a highly sensitive approach capable of measuring human topoisomerase 1 (TOP1) activity in single CD133 positive DLD-1 cells. The method termed On-Slide “Rolling circle Enhanced Enzyme Activity Detection (REEAD)” relies on the specific capture and lysis of CD133 positive cells on glass slides dual functionalized with anti-CD133 antibodies and a specific DNA primer. The On-Slide REEAD was demonstrated to be directly quantitative. Furthermore, the method allowed for the highly sensitive detection of TOP1 activity in single CD133 positive DLD-1 cells. The described protocol is expected to open for new possibilities in the single cell research, particularly for the investigations of chemoresistance of the cancer stem cells.

Original language	English
Title of host publication	Enzyme Activity in Single Cells
Editors	Nancy L. Allbritton, Michelle L. Kovarik
Number of pages	15
Publisher	Elsevier
Publication date	1 Jan 2019
Pages	43-57
ISBN (Print)	9780128170908
DOIs	https://doi.org/10.1016/bs.mie.2019.07.003
Publication status	Published - 1 Jan 2019

Series	Methods in Enzymology
Volume	628
ISSN	0076-6879

Keywords

Cancer
Cancer stem cells
Enzyme activity
Human topoisomerase 1
Microscopic slides
Rolling circle amplification
Single cell detection

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10.1016/bs.mie.2019.07.003

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title = "Enzymatic activity in single cells",

abstract = "With the increasing recognition of the importance in addressing cell-to-cell variations for the understanding of complex biological systems, single cell analyses are becoming increasingly important. Presented in this chapter is a highly sensitive approach capable of measuring human topoisomerase 1 (TOP1) activity in single CD133 positive DLD-1 cells. The method termed On-Slide “Rolling circle Enhanced Enzyme Activity Detection (REEAD)” relies on the specific capture and lysis of CD133 positive cells on glass slides dual functionalized with anti-CD133 antibodies and a specific DNA primer. The On-Slide REEAD was demonstrated to be directly quantitative. Furthermore, the method allowed for the highly sensitive detection of TOP1 activity in single CD133 positive DLD-1 cells. The described protocol is expected to open for new possibilities in the single cell research, particularly for the investigations of chemoresistance of the cancer stem cells.",

keywords = "Cancer, Cancer stem cells, Enzyme activity, Human topoisomerase 1, Microscopic slides, Rolling circle amplification, Single cell detection",

author = "Keller, {Josephine Geertsen} and Magnus Stougaard and Knudsen, {Birgitta R.}",

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AU - Stougaard, Magnus

AU - Knudsen, Birgitta R.

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N2 - With the increasing recognition of the importance in addressing cell-to-cell variations for the understanding of complex biological systems, single cell analyses are becoming increasingly important. Presented in this chapter is a highly sensitive approach capable of measuring human topoisomerase 1 (TOP1) activity in single CD133 positive DLD-1 cells. The method termed On-Slide “Rolling circle Enhanced Enzyme Activity Detection (REEAD)” relies on the specific capture and lysis of CD133 positive cells on glass slides dual functionalized with anti-CD133 antibodies and a specific DNA primer. The On-Slide REEAD was demonstrated to be directly quantitative. Furthermore, the method allowed for the highly sensitive detection of TOP1 activity in single CD133 positive DLD-1 cells. The described protocol is expected to open for new possibilities in the single cell research, particularly for the investigations of chemoresistance of the cancer stem cells.

AB - With the increasing recognition of the importance in addressing cell-to-cell variations for the understanding of complex biological systems, single cell analyses are becoming increasingly important. Presented in this chapter is a highly sensitive approach capable of measuring human topoisomerase 1 (TOP1) activity in single CD133 positive DLD-1 cells. The method termed On-Slide “Rolling circle Enhanced Enzyme Activity Detection (REEAD)” relies on the specific capture and lysis of CD133 positive cells on glass slides dual functionalized with anti-CD133 antibodies and a specific DNA primer. The On-Slide REEAD was demonstrated to be directly quantitative. Furthermore, the method allowed for the highly sensitive detection of TOP1 activity in single CD133 positive DLD-1 cells. The described protocol is expected to open for new possibilities in the single cell research, particularly for the investigations of chemoresistance of the cancer stem cells.

KW - Cancer

KW - Cancer stem cells

KW - Enzyme activity

KW - Human topoisomerase 1

KW - Microscopic slides

KW - Rolling circle amplification

KW - Single cell detection

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BT - Enzyme Activity in Single Cells

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PB - Elsevier

ER -

Enzymatic activity in single cells

Abstract

Keywords

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