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Related Experiment Videos

Temperature effect on CRISPR-Cas9 mediated genome editing.

Guanghai Xiang1, Xingying Zhang1, Chenrui An1

  • 1State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.

Journal of Genetics and Genomics = Yi Chuan Xue Bao
|April 17, 2017
PubMed
Summary

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Unlike other genome editing tools, CRISPR-Cas9 efficiency decreases with hypothermia. However, hyperthermia can enhance CRISPR-Cas9 editing in some cell lines by increasing enzyme activity and sgRNA expression.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Genome editing tools like ZFNs, TALENs, and CRISPR-Cas9 are crucial in molecular biology.
  • Previous research indicated hypothermia enhances ZFN and TALEN editing in mammalian cells.
  • The impact of temperature on CRISPR-Cas9 efficiency remained largely uncharacterized.

Purpose of the Study:

  • To investigate the effect of varying culture temperatures on CRISPR-Cas9 genome editing efficiency.
  • To compare CRISPR-Cas9's temperature sensitivity with ZFNs and TALENs.
  • To identify optimal temperature conditions for CRISPR-Cas9 applications.

Main Methods:

  • Culturing mammalian cells under hypothermic, normothermic (37°C), and hyperthermic conditions.
  • Utilizing CRISPR-Cas9 technology for genome editing.
Keywords:
CRISPR-Cas9Genome editingHyperthermiaMammalian cells

Related Experiment Videos

  • Quantifying genome editing efficiency at different temperatures.
  • Assessing enzyme activity and sgRNA expression levels at elevated temperatures.

    • Main Results:

    • Hypothermia significantly reduces CRISPR-Cas9 genome editing efficiency, contrasting with ZFNs and TALENs.
    • Hyperthermia enhances CRISPR-Cas9 editing efficiency in certain cell lines.
    • Increased enzyme activity and sgRNA expression at higher temperatures contribute to enhanced editing.

    Conclusions:

    • CRISPR-Cas9 exhibits distinct temperature sensitivity compared to ZFNs and TALENs.
    • Temperature optimization, particularly hyperthermia, can improve CRISPR-Cas9 efficacy in specific contexts.
    • Findings have broad implications for applying CRISPR-Cas9 across diverse species with varying optimal temperatures.