Department of Biology, The City College of New York, City University of New York (CUNY), New York, United States.
PhD Program in Biology, The Graduate Center of the City University of New York (CUNY), New York, United States.
Elife. 2020 Apr 29;9:e54279. doi: 10.7554/eLife.54279.
During vertebrate retinal development, subsets of progenitor cells generate progeny in a non-stochastic manner, suggesting that these decisions are tightly regulated. However, the gene-regulatory network components that are functionally important in these progenitor cells are largely unknown. Here we identify a functional role for the OTX2 transcription factor in this process. CRISPR/Cas9 gene editing was used to produce somatic mutations of OTX2 in the chick retina and identified similar phenotypes to those observed in human patients. Single cell RNA sequencing was used to determine the functional consequences OTX2 gene editing on the population of cells derived from OTX2-expressing retinal progenitor cells. This confirmed that OTX2 is required for the generation of photoreceptors, but also for repression of specific retinal fates and alternative gene regulatory networks. These include specific subtypes of retinal ganglion and horizontal cells, suggesting that in this context, OTX2 functions to repress sister cell fate choices.
在脊椎动物视网膜发育过程中，祖细胞亚群以非随机的方式产生后代，这表明这些决定受到严格调控。然而，在这些祖细胞中具有功能重要性的基因调控网络组件在很大程度上尚不清楚。在这里，我们确定了 OTX2 转录因子在这个过程中的功能作用。使用 CRISPR/Cas9 基因编辑在鸡视网膜中产生 OTX2 的体细胞突变，并鉴定出与在人类患者中观察到的相似表型。单细胞 RNA 测序用于确定 OTX2 基因编辑对源自 OTX2 表达的视网膜祖细胞的细胞群体的功能后果。这证实了 OTX2 不仅对于光感受器的产生是必需的，而且对于抑制特定的视网膜命运和替代基因调控网络也是必需的。这些网络包括特定类型的视网膜神经节细胞和水平细胞，这表明在这种情况下，OTX2 起抑制姐妹细胞命运选择的作用。
