Factors Affecting Genome Editing Using Crispr/Cas9 In Mouse Model

CRISPR/Cas9 joined genome editing family and became a powerful tool for cancer researches. While CRISPR is effective for generating knock-out, knock-in or point mutation animal models, the protocols and success rates to generate CRISPR mouse model vary significantly among different laboratories. We examined a series of parameters for generating various types of mouse models using CRISPR/Cas9. The efficiency and toxicity of gRNA, Cas9 mRNA and donor concentration, insert size and injection methods were thoroughly analyzed. We also evaluated strategies generating conditional knock-out models using CRISPR. The optimized protocols for successful mouse model generation for knock-in, knock-out, point mutation and conditional knock-out using CRISPR will be proposed. Citation Format: Qi Zheng, Ling-Jie Kong, Huanyu Jin, Jinling Li, Ruby Yanru Chen-Tsai. Factors affecting genome editing using CRISPR/Cas9 in mouse model. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 663.