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Application of digital PCR for assessing DNA fragmentation in cytotoxicity response

Jeongran Han1, Ji Youn Lee2, Young-Kyung Bae2

  • 1Center for Bioanalysis, Korea Research Institute of Standards and Science, 267 Gajeong-ro, Yuseong-gu, Daejeon, Republic of Korea; Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon, Republic of Korea.

Biochimica Et Biophysica Acta. General Subjects
|May 10, 2019
PubMed
Abstract

Insights

This study introduces novel digital PCR (dPCR) assays to measure cell death by quantifying DNA fragmentation. The RP fragmentation index provides a precise method for assessing cytotoxicity in human cell lines.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biotechnology

Background:

  • Regulated cell death is crucial in biological processes.
  • Existing methods quantify cell death due to apoptosis or necrosis.
  • Cytotoxicity assays often use human cell lines sensitive to anti-cancer agents.

Purpose of the Study:

  • To develop novel digital PCR (dPCR) assays for quantitative measurement of cell death.
  • To establish a new metric, the RP fragmentation index, for assessing DNA fragmentation.
  • To evaluate the efficacy of dPCR assays in measuring cytotoxicity.

Main Methods:

  • Designed novel digital PCR (dPCR) assays to quantify targets of varying sizes within the RNase P (RP) gene locus.
  • Utilized the ratio of short to long target copy numbers to determine the degree of DNA fragmentation.
  • Calculated the RP fragmentation index based on DNA fragmentation levels.

Main Results:

  • The developed dPCR assays precisely quantify DNA fragmentation.
  • The RP fragmentation index accurately reflects the degree of cell death.
  • dPCR assays demonstrate high sensitivity in measuring cytotoxicity.

Conclusions:

  • The RP fragmentation index is a valid indicator for measuring cell death.
  • dPCR assays offer a precise method for assessing DNA fragmentation and cytotoxicity.
  • These assays provide a quantitative measure of cell death in response to cytotoxic agents.
Keywords:
CytotoxicityDNA fragmentationDigital PCRRegulated cell death

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