Systematic evaluation of CRISPR-Cas systems reveals higher efficiency of Cpf1 nucleases in HDR-mediated precise genome editing

• Main Author: Xing, Heyun
• Other Authors: Tan Meng How
• Format: Theses and Dissertations
• Language: English
• Published: 2018
• Subjects: DRNTU::Engineering::Bioengineering
• Online Access: http://hdl.handle.net/10356/75292
• Institution: Nanyang Technological University

Background: CRISPR/Cas has the advantages of low cost, easy production, rapid and high efficiency, and low off-target rate. It has quickly become popular in laboratories around the world, and has become an effective tool in scientific research, medical care, and other fields. One of the hottest technologies in the field of molecular biology today. Results: In this paper, we systematically assessed the ability of different CRISPR-Cas systems to HDR-mediated genome editing in human cells. We targeted numerous genomic loci with matched spacers or matched seeds to obtain a clearer and fairer picture of how the various Cas enzymes compared against one another. Based on experiments data, we demonstrat that Cpf1 nucleases are more efficient in genome edits when ssODNs are used as donor templates. And we derive a set of rules that others may follow to select an appropriate CRISPR-Cas system for their experiments. Conclusions: Collectively, our work revealed several novel and unexpected findings on the performance of different CRISPR-Cas systems and will help guide the design of future genome engineering studies.