@article {65078,
	title = {Unlimited genetic switches for cell-type-specific manipulation.},
	journal = {Neuron},
	volume = {104},
	year = {2019},
	month = {2019 Oct 23},
	pages = {227-38},
	abstract = {<p>Gaining independent genetic access to discrete cell types is critical to interrogate their biological functions as well as to deliver precise gene therapy. Transcriptomics has allowed us to profile cell populations with extraordinary precision, revealing that cell types are typically defined by a unique combination of genetic markers. Given the lack of adequate tools to target cell types based on multiple markers, most cell types remain inaccessible to genetic manipulation. Here we present CaSSA, a platform to create unlimited genetic switches based on CRISPR/Cas9 (Ca) and the DNA repair mechanism known as single-strand annealing (SSA). CaSSA allows engineering of independent genetic switches, each responding to a specific gRNA. Expressing multiple gRNAs in specific patterns enables multiplex cell-type-specific manipulations and combinatorial genetic targeting. CaSSA is a new genetic tool that conceptually works as an unlimited number of recombinases and will facilitate genetic access to cell types in diverse organisms.</p>
},
	doi = {https://doi.org/10.1016/j.neuron.2019.07.005},
	author = {Garcia-Marques, Jorge and Yang, Ching-Po and Isabel Espinosa Medina and Mok, Kent and Koyama, Minoru and Lee, Tzumin}
}
@article {48435,
	title = {Stem cell-intrinsic, seven-up-triggered temporal factor gradients diversify intermediate neural progenitors.},
	journal = {Current Biology : CB},
	volume = {27},
	year = {2017},
	month = {2017 Apr 10},
	pages = {1303-13},
	abstract = {<p>Building a sizable, complex brain requires both cellular expansion and diversification. One mechanism to achieve these goals is production of multiple transiently amplifying intermediate neural progenitors (INPs) from a single neural stem cell. Like mammalian neural stem cells, Drosophila type II neuroblasts utilize INPs to produce neurons and glia. Within a given lineage, the consecutively born INPs produce morphologically distinct progeny, presumably due to differential inheritance of temporal factors. To uncover the underlying temporal fating mechanisms, we profiled type II neuroblasts{\textquoteright} transcriptome across time. Our results reveal opposing temporal gradients of Imp and Syp RNA-binding proteins (descending and ascending, respectively). Maintaining high Imp throughout serial INP production expands the number of neurons and glia with early temporal fate at the expense of cells with late fate. Conversely, precocious upregulation of Syp reduces the number of cells with early fate. Furthermore, we reveal that the transcription factor Seven-up initiates progression of the Imp/Syp gradients. Interestingly, neuroblasts that maintain initial Imp/Syp levels can still yield progeny with a small range of early fates. We therefore propose that the Seven-up-initiated Imp/Syp gradients create coarse temporal windows within type II neuroblasts to pattern INPs, which subsequently undergo fine-tuned subtemporal patterning.</p>
},
	issn = {1879-0445},
	doi = {10.1016/j.cub.2017.03.047},
	author = {Ren, Qingzhong and Yang, Ching-Po and Liu, Zhiyong and Sugino, Ken and Mok, Kent and He, Yisheng and Ito, Masayoshi and Nern, Aljoscha and Otsuna, Hideo and Lee, Tzumin}
}
