Targeted Mutagenesis of Beta-Lactoglobulin Gene in Caprine Fetal Fibroblasts by Context-Dependent Assembly Zinc-Finger Nucleases - Open Access Library Journal

OALibJ > Vol.3 No.7, July 2016

Targeted Mutagenesis of Beta-Lactoglobulin Gene in Caprine Fetal Fibroblasts by Context-Dependent Assembly Zinc-Finger Nucleases ()

HTML XML

Download as PDF (Size: 423KB) PP. 1-8

DOI: 10.4236/oalib.1102813 730 Downloads 1,413 Views Citations

Author(s)

Yuguo Yuan^1,2, Yong Cheng¹, Jinyu Wang^2*, Qiuling Peng^3*

Affiliation(s)

¹College of Veterinary Medicine, Yangzhou University/Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China.
²College of Animal Science and Technology, Yangzhou University, Yangzhou, China.
³Department of Biological and Chemical Engineering, Yichun University, Yichun, China.

ABSTRACT

Targeted mutagenesis by zinc-finger nucleases (ZFNs) can be used to generate knock-out mammalian cell lines with high efficiency. A number of different methods have been developed for the design and assembly of gene-specific ZFNs, making them easily accessible to researchers. In this study, we used ZFNs assembled through the CoDA (context-dependent assembly) platforms to generate mutant caprine fetal fibroblasts cells for the BLG gene. ZFN plasmid was introduced into the caprine fetal fibroblasts cell by electroporation. ZFN-induced cleavage of the target sequence was confirmed by the Surveyor nuclease assay analysis. Sequence analysis revealed that ZFN-induced mutations such as base insertion, deletion, or substitution were generated in the ZFN cleavage site of BLG. The simplicity and efficacy of CoDA will enable broad application of ZFN technology. This technique could be used with homologous arm, which may target foreign genes into the BLG locus at higher efficiency.

KEYWORDS

Context-Dependent Assembly, Zinc Finger Nucleases, Beta-Lactoglobulin, Gene Knockout, Goat

Share and Cite:

Yuan, Y. , Cheng, Y. , Wang, J. and Peng, Q. (2016) Targeted Mutagenesis of Beta-Lactoglobulin Gene in Caprine Fetal Fibroblasts by Context-Dependent Assembly Zinc-Finger Nucleases. Open Access Library Journal, 3, 1-8. doi: 10.4236/oalib.1102813.

Journals Menu

Follow SCIRP

	+1 323-425-8868
	customer@scirp.org
	+86 18163351462(WhatsApp)
	1655362766

	Paper Publishing WeChat

Journals Menu

Home

About SCIRP

Service

Policies