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Single fluorescent protein-based Ca2+ sensors with increased dynamic range.

Ekaterina A Souslova1, Vsevolod V Belousov, John G Lock

  • 1Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, RAS, Miklukho-Maklaya 16/10, Moscow, Russia. souslova@gmail.com <souslova@gmail.com>

BMC Biotechnology
|July 3, 2007
PubMed
Summary
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New genetically encoded calcium sensors, Case12 and Case16, offer enhanced brightness and dynamic range for cellular monitoring. These fluorescent protein sensors provide superior performance for tracking calcium levels in living cells and organisms.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Genetically encoded sensors based on green fluorescent protein (GFP)-like proteins are widely used for monitoring cellular analytes.
  • Calcium (Ca2+) sensors are crucial, with existing types based on Fluorescence Resonance Energy Transfer (FRET) or single circularly permuted fluorescent proteins (cpFP).

Purpose of the Study:

  • To develop advanced cpFP-based Ca2+ sensors with improved spectral properties and fluorescent responses.
  • To create novel Ca2+ indicators with high brightness and expanded dynamic range.

Main Methods:

  • Generation of a set of cpFP-based indicators derived from existing sensor knowledge.
  • Characterization of spectral properties and Ca2+ concentration-dependent fluorescent responses.
  • Demonstration of sensor utility in monitoring cellular Ca2+ dynamics.

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Main Results:

  • Developed two variants, Case12 and Case16, exhibiting high brightness and superior dynamic range.
  • Achieved up to 12-fold (Case12) and 16.5-fold (Case16) increases in green fluorescence.
  • Successfully monitored Ca2+ responses, such as to glutamate treatment in cortical neurons.

Conclusions:

  • Case12 and Case16 offer expanded dynamic range and high brightness, making them valuable research tools.
  • These sensors are suitable for both scientific studies and high-throughput screening assays.
  • The pH-stability of these sensors further enhances their utility in biological research.