The Laboratory of Chromatin
Biology and Cancer Epigenetics

Lu Yang, Anthony K N Chan, Kazuya Miyashita, Christopher D Delaney, Xi Wang, Hongzhi Li, Sheela Pangeni Pokharel, Sandra Li, Mingli Li, Xiobao Xu, Wei Lu, Qiao Liu, Nicole Mattson, Kevin Yining Chen, Jinhui Wang, Yate-Ching Yuan, David Horne, Steven T Rosen, Yadira Soto-Feliciano, Zhaohui Feng, Takayiki Hoshii, Gang Xiao, Markus Muschen, Jianjun Chen, Scott A Armstrong, Chen-Wei Chen

Identification of novel functional domains and characterization of detailed regulatory mechanisms in cancer-driving genes is critical for advanced cancer therapy. To date, CRISPR gene editing has primarily been applied to defining the role of individual genes. Recently, high-density mutagenesis via CRISPR tiling of gene-coding exons has been demonstrated to identify functional regions in genes. Furthermore, breakthroughs in combining CRISPR library screens with single-cell droplet RNA sequencing (sc-RNAseq) platforms have revealed the capacity to monitor gene expression changes upon genetic perturbations at single-cell resolution. Here, we present “sc-Tiling,” which integrates a CRISPR gene-tiling screen with single-cell transcriptomic and protein structural analyses. Distinct from other reported single-cell CRISPR screens focused on observing gene function and gene-to-gene/enhancer-to-gene regulation, sc-Tiling enables the capacity to identify regulatory mechanisms within a gene-coding region that dictate gene activity and therapeutic response.