Cloning and base editing of GFP transgenic rhesus monkey and off-target analysis-Reference-Cited by-同舟云学术

Cloning and base editing of GFP transgenic rhesus monkey and off-target analysis

Published:2022-07-22 Issue:29 Volume:8 Page:
ISSN:2375-2548
Container-title:Science Advances
language:en
Short-container-title:Sci. Adv.

Author:

Kang Yu¹²³^ORCID,Dai Shaoxing¹³^ORCID,Zeng Yuqiang¹³^ORCID,Wang Fang¹³^ORCID,Yang Pengpeng¹³,Yang Zhaohui¹³^ORCID,Pu Youwei¹³^ORCID,Li Zifan¹³,Chen Xinglong¹³,Tian Baohong¹³,Si Wei¹³,Ji Weizhi¹²³^ORCID,Niu Yuyu¹²³^ORCID

Affiliation:

1. State Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China.

2. Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, Yunnan 650500, China.

3. Yunnan Key Laboratory of Primate Biomedical Research, Kunming, Yunnan 650500, China.

Abstract

We report the cloning of a 12-year-old transgenic green fluorescent protein (GFP) monkey by somatic cell nuclear transfer (SCNT) and base editing of the embryos, accompanied with safety evaluation of adenine base editors (ABEs). We first show the ability of ABEmax to silence GFP through A-to-G editing of the GFP sequence in 293T cells. Subsequently, using donor cells from a monkey expressing GFP, we have successfully generated 207 ABEmax-edited (SCNT-ABE) and 87 wild-type (SCNT) embryos for embryo transfer, genotyping, and genome and transcriptome analysis. SCNT-ABE and SCNT embryos are compared for off-target analysis without the interference of genetic variants using a new method named as OA-SCNT. ABEmax does not induce obvious off-target DNA mutations but induces widespread off-target RNA mutations, 35% of which are exonic, in edited monkey embryos. These results provide important references for clinical application of ABE.

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

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