The growth and reproduction performance of TALEN-mediated β-lactoglobulin-knockout bucks.
With the technological development of several engineered endonucleases (EENs), such as zinc-finger nucleases, transcription activator-like effector nucleases (TALENs) and CRISPR/Cas9, gene targeting by homologous recombination has been efficiently improved to generate site-specifically genetically modified livestock. However, few studies have been done to investigate the health and fertility of these animals. The purpose of the present study is to investigate if gene targeting events and a recloning procedure would affect the production traits of EEN-mediated gene targeted bucks. TALEN-mediated β-lactoglobulin (BLG) gene mono-allelic knockout (BLG (+/-)) goats and bi-allelic knockout (BLG (-/-)) buck produced by using sequential gene targeting combined with recloning in fibroblasts from BLG (+/-) buck were used to evaluate their health and fertility. Birth weight and postnatal growth of BLG (+/-) bucks were similar to the wild-type goats. None of the parameters for both fresh and frozen-thawed semen quality were significantly different in BLG (+/-) or BLG (-/-) bucks compared to their corresponding comparators. In vitro fertilization (IVF) test revealed that the proportion of IVF oocytes developing to the blastocyst stage was identical among BLG (+/-), BLG (-/-) and wild-type bucks. Conception rates of artificial insemination were respectively 42.3, 38.0 and 42.6 % for frozen-thawed semen from the BLG (+/-), BLG (-/-) and wild-type bucks. In addition, germline transmission of the targeted BLG modification was in accordance with Mendelian rules. These results demonstrated that the analyzed growth and reproductive traits were not impacted by targeting BLG gene and recloning, implicating the potential for dairy goat breeding of BLG (+/-) and BLG (-/-) bucks.
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China.
Hengtao
H
Ge
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China.
Chenchen
C
Cui
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China.
Jun
J
Liu
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China.
Yan
Y
Luo
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China.
Fusheng
F
Quan
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China. yapingjin@163.com.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China. yapingjin@163.com.
Yaping
Y
Jin
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China. yongzh1956@126.com.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China. yongzh1956@126.com.
Yong
Y
Zhang
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000254:growth & development; D001947:Breeding; D064113:CRISPR-Cas Systems / Q000235:genetics; D005298:Fertility / Q000235:genetics; D055786:Gene Knockout Techniques; D018390:Gene Targeting / Q000379:methods; D006041:Goats / Q000235:genetics* / Q000254:growth & development; D059765:Homologous Recombination / Q000235:genetics*; D007782:Lactoglobulins / Q000235:genetics*; D008892:Milk / Q000378:metabolism; D012098:Reproduction / Q000235:genetics; D000069896:Transcription Activator-Like Effector Nucleases / Q000235:genetics
D007782:Lactoglobulins; D000069896:Transcription Activator-Like Effector Nucleases
Gene targeting; Goat; Recloning; Reproductive performance