Volume 26, No 5, May 2016

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 26 Issue 5, May 2016: 633-636

LETTERS TO THE EDITOR

One-step homozygosity in precise gene editing by an improved CRISPR/Cas9 system

Pei Zhao^1,2,*, Zhe Zhang^1,*, Xiaoying Lv³, Xiang Zhao¹, Yuji Suehiro⁴, Yinan Jiang³, Xinquan Wang³, Shohei Mitani⁴, Haipeng Gong³ and Ding Xue^1,5

¹School of Life Sciences & Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China
²Tsinghua University-Peking University Joint Center for Life Science, Beijing 100084, China
³School of Life Sciences, Tsinghua University, Beijing 100084, China
⁴Department of Physiology, Tokyo Women's Medical University, School of Medicine, and Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Tokyo 162-8666, Japan
⁵Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309, USA Correspondence: Ding Xue,(ding.xue@colorado.edu)

The CRISPR/Cas9 gene editing method has been successfully applied to modify genomes in many organisms1. However, several critical issues remain unresolved and have become major hurdles for its broad applications1. First, editing efficiency varies widely at different genetic loci and some targeted sites are resistant to editing for unknown reasons. Even for the same gene, editing efficiency differs greatly at different positions. Second, generation of undesirable insertions or deletions (InDels) at the target sites constitutes a major issue in precise genome editing and gene therapy. Third, one-step homozygosity in precise gene editing is extremely rare and is highly desirable for genetic and functional analyses, especially in systems where genetic manipulations are not possible or are tedious and time-consuming, such as cultured cells and mammals.

10.1038/cr.2016.46

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