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Efficient photoactivatable Dre recombinase for cell type-specific spatiotemporal control of genome engineering in the mouse.

Huiying LiQiansen ZhangYiran GuYingyin WuYamei WangLiren WangShijie FengYaqiang HuYansen ZhengYongmei LiHaifeng YeBin ZhouLongnian LinMingyao LiuHuaiyu YangDali Li
Published in: Proceedings of the National Academy of Sciences of the United States of America (2020)
Precise genetic engineering in specific cell types within an intact organism is intriguing yet challenging, especially in a spatiotemporal manner without the interference caused by chemical inducers. Here we engineered a photoactivatable Dre recombinase based on the identification of an optimal split site and demonstrated that it efficiently regulated transgene expression in mouse tissues spatiotemporally upon blue light illumination. Moreover, through a double-floxed inverted open reading frame strategy, we developed a Cre-activated light-inducible Dre (CALID) system. Taking advantage of well-defined cell-type-specific promoters or a well-established Cre transgenic mouse strain, we demonstrated that the CALID system was able to activate endogenous reporter expression for either bulk or sparse labeling of CaMKIIα-positive excitatory neurons and parvalbumin interneurons in the brain. This flexible and tunable system could be a powerful tool for the dissection and modulation of developmental and genetic complexity in a wide range of biological systems.
Keyphrases
  • poor prognosis
  • genome wide
  • single cell
  • crispr cas
  • binding protein
  • minimally invasive
  • multiple sclerosis
  • dna methylation
  • cell therapy
  • long non coding rna
  • light emitting