Nucleic acid engineering is a group of technologies that can change the script of life, such as genome and transcriptome, enabling a better understanding of human genomics, and can be developed as genetic medicines to treat diseases. Currently, there are three major types of gene editing approaches, including nuclease editing, base editing, and prime editing. Yet, the therapeutic applications of those technologies are still facing unmet needs. Although gene editing strategies have been demonstrated for monogenic disorders, such as sickle cell anemia, cystic fibrosis, Huntington disease, and Duchenne muscular dystrophy, the genetic or cellular treatment of polygenic disorder that caused by combined dysfunctions of more than one gene, such as coronary heart disease, diabetes, cancer, and neurological diseases, still needs to be well developed. How to further advance those technologies, or to develop next-generation gene editing tools that can perfectly address the emerging challenges from real-life medical issues? To approach a solution of the previous question, I focused my research on six topics: 1) Enable multiplex precision genome engineering with minimal delivery size, 2) Expand the type and number of genetic perturbations, 3) Manipulate cellular endogenous biological mechanisms to advance the performance of molecular tools, 4) Engineer the key components of molecular tools with improved activity, 5) Demonstrate therapeutic genome engineering on disease-relevant genes, 6) Deliver the therapeutic genetic payload using virial and non-viral approaches.