CRISPR Start-Loss: A Novel and Practical Alternative for Gene Silencing through Base-Editing-Induced Start Codon Mutations

• Main Authors: Siyu Chen, Wanhua Xie, Zhiquan Liu, Huanhuan Shan, Mao Chen, Yuning Song, Hao Yu, Liangxue Lai, Zhanjun Li
• Format: Article
• Language: English
• Published: Elsevier 2020-09-01
• Series: Molecular Therapy: Nucleic Acids
• Subjects: CRISPR start-loss; start codon; loss-of-function; Fgf5; Otc; rabbits
• Online Access: http://www.sciencedirect.com/science/article/pii/S2162253120302262
• ISSN: 2162-2531

CRISPR-Cas9-mediated gene knockout and base-editing-associated induction of STOP codons (iSTOP) have been widely used to exterminate the function of a coding gene, while they have been reported to exhibit side effects. In this study, we propose a novel and practical alternative method referred to as CRISPR Start-Loss (CRISPR-SL), which eliminates gene expression by utilizing both adenine base editors (ABEs) and cytidine base editors (CBEs) to disrupt the initiation codon (ATG). CRISPR-SL has been verified to be a feasible strategy on the cellular and embryonic levels (mean editing efficiencies up to 30.67% and 73.50%, respectively) and in two rabbit models mimicking Otc deficiency (Otc gene) and long hair economic traits (Fgf5 gene).