Infrared Laser-Mediated Gene Induction at the Single-Cell Level in the Regenerating Tail of Xenopus laevis Tadpoles

Cold Spring Harbor Protocols
Riho HasugataHitoshi Yokoyama

Abstract

We describe a precise and reproducible gene-induction method in the amphibian, Xenopus laevis Tetrapod amphibians are excellent models for studying the mechanisms of three-dimensional organ regeneration because they have an exceptionally high regenerative ability. However, spatial and temporal manipulation of gene expression has been difficult in amphibians, hindering studies on the molecular mechanisms of organ regeneration. Recently, however, development of a Xenopus transgenic system with a heat-shock-inducible gene has enabled the manipulation of specific genes. Here, we applied an infrared laser-evoked gene operator (IR-LEGO) system to the regenerating tail of Xenopus tadpoles. In this method, a local heat shock by laser irradiation induces gene expression at the single-cell level. After amputation, Xenopus tadpoles regenerate a functional tail, including spinal cord. The regenerating tail is flat and transparent enabling the targeting of individual cells by laser irradiation. In this protocol, a single neural progenitor cell in the spinal cord of the regenerating tail is labeled with heat-shock-inducible green fluorescent protein (GFP). Gene induction at the single-cell level provides a method for rigorous cell-lineage tr...Continue Reading

References

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Dec 17, 2008·Nature Methods·Yasuhiro KameiShunsuke Yuba
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Mar 29, 2013·Nature Communications·Atsuko ShimadaHiroyuki Takeda

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Citations

Jul 31, 2019·Plant & Cell Physiology·Dukhyun HwangShuh-Ichi Nishikawa
Jun 28, 2018·Development, Growth & Differentiation·Hitoshi YokoyamaKoji Tamura
Feb 18, 2020·Biophysics Reviews·Shuji AkiyamaYoshihiro Kubo
Mar 31, 2021·Cold Spring Harbor Protocols·Paula G SlaterJuan Larraín

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