Biology Department, Boston University, Boston, 02215, USA.
Biology Department, Massachusetts Institute of Technology, Cambridge, 02139, USA.
Nat Commun. 2023 Jun 10;14(1):3435. doi: 10.1038/s41467-023-39200-2.
Transcription and splicing are intrinsically coupled. Alternative splicing of internal exons can fine-tune gene expression through a recently described phenomenon called exon-mediated activation of transcription starts (EMATS). However, the association of this phenomenon with human diseases remains unknown. Here, we develop a strategy to activate gene expression through EMATS and demonstrate its potential for treatment of genetic diseases caused by loss of expression of essential genes. We first identified a catalog of human EMATS genes and provide a list of their pathological variants. To test if EMATS can be used to activate gene expression, we constructed stable cell lines expressing a splicing reporter based on the alternative splicing of motor neuron 2 (SMN2) gene. Using small molecules and antisense oligonucleotides (ASOs) currently used for treatment of spinal muscular atrophy, we demonstrated that increase of inclusion of alternative exons can trigger an activation of gene expression up to 45-fold by enhancing transcription in EMATS-like genes. We observed the strongest effects in genes under the regulation of weak human promoters located proximal to highly included skipped exons.
转录和剪接是内在偶联的。通过最近描述的一种称为外显子介导的转录起始激活（exon-mediated activation of transcription starts，EMATS）的现象，内部外显子的可变剪接可以精细调节基因表达。然而，这种现象与人类疾病的关联尚不清楚。在这里，我们开发了一种通过 EMATS 激活基因表达的策略，并证明了其在治疗因必需基因表达缺失引起的遗传疾病方面的潜力。我们首先鉴定了人类 EMATS 基因的目录，并提供了其病理性变体的列表。为了测试 EMATS 是否可用于激活基因表达，我们构建了表达基于运动神经元 2（SMN2）基因可变剪接的剪接报告基因的稳定细胞系。使用目前用于治疗脊髓性肌萎缩症的小分子和反义寡核苷酸（ASO），我们证明增加可变外显子的包含可以通过在 EMATS 样基因中增强转录来触发基因表达高达 45 倍的激活。我们在靠近高度包含的跳过外显子的弱人启动子调控下的基因中观察到最强的效应。
