Next Article in Journal
Identification of Novel Circular RNAs of the Human Protein Arginine Methyltransferase 1 (PRMT1) Gene, Expressed in Breast Cancer Cells
Previous Article in Journal
Mapping-by-Sequencing Reveals Genomic Regions Associated with Seed Quality Parameters in Brassica napus
Previous Article in Special Issue
Application of CRISPR/Cas Technology in Spermatogenesis Research and Male Infertility Treatment
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Genes
Volume 13
Issue 7
10.3390/genes13071132
genes-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Efficient Editing of the ZBED6-Binding Site in Intron 3 of IGF2 in a Bovine Model Using the CRISPR/Cas9 System

by
Huiying Zou
1,†,
Dawei Yu
1,†,
Shun Yao
1,†,
Fangrong Ding
2,
Junliang Li
1,
Ling Li
2,
Xue Li
2,
Shanjiang Zhao
1,
Yunwei Pang
1,
Haisheng Hao
1,
Weihua Du
1,
Xueming Zhao
1,
Yunping Dai
2,* and
Huabin Zhu
1,*
1
Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
2
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Genes 2022, 13(7), 1132; https://doi.org/10.3390/genes13071132
Submission received: 22 April 2022 / Revised: 16 May 2022 / Accepted: 16 May 2022 / Published: 24 June 2022
(This article belongs to the Special Issue Application of Transgenic Technology in Animal Breeding)
Browse Figures
Versions Notes

Abstract

Background: Insulin-like growth factor 2 is a growth-promoting factor that plays an important role in the growth and development of mammals. A nucleotide substitution in intron 3 of IGF2—which disrupts the ZBED6-binding site—affects muscle mass, organ size, and fat deposition in pigs. The ZBED6-binding site is also conserved in cattle. Methods: In the present study, we introduced mutations in the ZBED6-binding site in intron3 of IGF2 in bovine fetal fibroblasts using the CRISPR/Cas9 system, and investigated the effect of disruption of ZBED6 binding on IGF2 expression. Results: Eleven biallelic-mutant single-cell clones were established, three of which contained no foreign DNA residues. Single-cell clones 93 and 135 were used to produce cloned embryos. Dual-luciferase reporter assay in C2C12 cells demonstrated that the mutation in the ZBED6-binding site increases the promoter 3 activity of bovine IGF2. A total of 49 mutant cloned embryos were transplanted into surrogate cows. Unfortunately, all cloned embryos died before birth. IGF2 was found to be hypomethylated in the only fetus born (stillborn), which may have been due to the incomplete reprogramming. Conclusions: We efficiently constructed IGF2-edited cell lines and cloned embryos, which provided a theoretical basis and experimental materials for beef cattle breeding.
Keywords: CRISPR/Cas9 system; gene editing; insulin-like growth factor 2; ZBED6-binding site; cattle CRISPR/Cas9 system; gene editing; insulin-like growth factor 2; ZBED6-binding site; cattle

Share and Cite

MDPI and ACS Style

Zou, H.; Yu, D.; Yao, S.; Ding, F.; Li, J.; Li, L.; Li, X.; Zhao, S.; Pang, Y.; Hao, H.; et al. Efficient Editing of the ZBED6-Binding Site in Intron 3 of IGF2 in a Bovine Model Using the CRISPR/Cas9 System. Genes 2022, 13, 1132. https://doi.org/10.3390/genes13071132

AMA Style

Zou H, Yu D, Yao S, Ding F, Li J, Li L, Li X, Zhao S, Pang Y, Hao H, et al. Efficient Editing of the ZBED6-Binding Site in Intron 3 of IGF2 in a Bovine Model Using the CRISPR/Cas9 System. Genes. 2022; 13(7):1132. https://doi.org/10.3390/genes13071132

Chicago/Turabian Style

Zou, Huiying, Dawei Yu, Shun Yao, Fangrong Ding, Junliang Li, Ling Li, Xue Li, Shanjiang Zhao, Yunwei Pang, Haisheng Hao, and et al. 2022. "Efficient Editing of the ZBED6-Binding Site in Intron 3 of IGF2 in a Bovine Model Using the CRISPR/Cas9 System" Genes 13, no. 7: 1132. https://doi.org/10.3390/genes13071132

APA Style

Zou, H., Yu, D., Yao, S., Ding, F., Li, J., Li, L., Li, X., Zhao, S., Pang, Y., Hao, H., Du, W., Zhao, X., Dai, Y., & Zhu, H. (2022). Efficient Editing of the ZBED6-Binding Site in Intron 3 of IGF2 in a Bovine Model Using the CRISPR/Cas9 System. Genes, 13(7), 1132. https://doi.org/10.3390/genes13071132

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop