Construction of a system for single-cell transgene induction in Caenorhabditis elegans using a pulsed infrared laser.
The spatial and temporal control of transgene expression is an important tool in Caenorhabditis elegans biology. We previously described a method for evoking gene expression in arbitrary cells by using a focused pulsed infrared laser to induce a heat shock response (Churgin et al., 2013). Here we describe detailed methods for building and testing a system for performing single-cell heat shock. Steps include setting up the laser and associated components, coupling the laser beam to a microscope, and testing heat shock protocols. All steps can be carried out using readily available off-the-shelf components.
Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, PA 19104, United States.
Matthew A
MA
Churgin
Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, PA 19104, United States.
Liping
L
He
Department of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States.
John I
JI
Murray
Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, PA 19104, United States; Department of Physics, Korea University, Anam-dong, Seongbuk-gu, Seoul 136-701, South Korea. Electronic address: fangyen@seas.upenn.edu.
Christopher
C
Fang-Yen
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D017173:Caenorhabditis elegans / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental / Q000528:radiation effects*; D018869:Heat-Shock Response / Q000528:radiation effects*; D007834:Lasers; D011401:Promoter Regions, Genetic / Q000528:radiation effects; D059010:Single-Cell Analysis / Q000379:methods*; D019076:Transgenes
C. elegans; Heat shock; Lasers; Transgenes