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.
Cite this paper
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, e2813. doi: http://dx.doi.org/10.4236/oalib.1102813.
Provost, F.L., Lillico, S., Passet, B., Young, R., Whitelaw, B. and
Vilotte,
J.L. (2009) Zinc Finger Nuclease Technology Heralds a New Era in Mammalian
Transgenesis. Trends Biotechnology, 28,
134-141. http://dx.doi.org/10.1016/j.tibtech.2009.11.007
Rémy, S., Tesson, L.S., Ménoret, C., Usal, A., Scharenberg, M. and
Anegon, S. (2010) Zinc-Finger Nucleases: A Powerful Tool for Genetic
Engineering of Animals. Transgenic
Research, 19, 363-371. http://dx.doi.org/10.1007/s11248-009-9323-7
Lei, Y., Lee, C. L., Joo, K., Zarzar, J., Liu, Y., Dai, B., Fox, V.
and Wang, P. (2011) Gene Editing of Human Embryonic Stem Cells via an Engineered
Baculoviral Vector Carrying Zinc-Finger Nucleases. Molecular therapy, 19, 942-950. http://dx.doi.org/10.1038/mt.2011.12
Meyera, M., Angelis, M.H., Wursta, W. and Kühn,
R. (2010) Gene Targeting by Homologous Recombination in Mouse Zygotes Mediated
by Zinc-Finger Nucleases. Proceedings of
the National Academy of Sciences of the USA, 107, 15022-15026. http://dx.doi.org/10.1073/pnas.1009424107
Hauschilda, J., Petersena, B., Santiagob, Y.,
Queissera, A.L., Carnwatha, J.W., Lucas-Hahna, A., Zhang, L., Meng, X.,
Gregory, P.D., Schwinzerd, R., Costb, G.J. and Niemanna, H. (2011) Efficient Generation
of a Biallelic Knockout in Pigs Using Zinc-Finger
Nucleases. Proceedings of the National
Academy of Sciences of the USA, 108, 2013-2017. http://dx.doi.org/10.1073/pnas.1106422108
Hauschild-Quintern, J., Petersen, B., Cost,
G.J. and Niemann. H. (2012) Gene Knockout and Knockin by Zinc-Finger Nucleases:
Current Status and Perspectives. Cellular
Molecular Life Sciences, 70, 2969-2983. http://dx.doi.org/10.1007/s00018-012-1204-1
Chen, S., Oikonomou, G., Chiu, C.N., Niles,
B.J., Liu, J., Lee, D.A., Antoshechkin, I. and Prober, D.A. (2013) A large-Scale in Vivo Analysis Reveals That TALENs Are
Significantly More Mutagenic than ZFNs Generated Using Context-Dependent Assembly. Nucleic Acids Research, 41, 2769-2778. http://dx.doi.org/10.1093/nar/gks1356
Sood, R., Carrington, B., Bishop, K., Jones, M., Rissone, A., Candotti, F., Chandrasekharappa, S.C. and Liu, P. (2013) Efficient Methods for Targeted
Mutagenesis in Zebrafish Using Zinc-Finger Nucleases: Data from Targeting of Nine
Genes Using CompoZr or CoDA ZFNs. PloS
One, 8, e57239. http://dx.doi.org/10.1371/journal.pone.0057239
Kwon, D. N., Lee, K., Kang, M. J., Choi, Y.J.,
Park, C., Jeffrey, J., Brown, A.N., Kim, J.H., Samue, M., Mao, J., Park, K.W.,
Murphy, C.N., Prather, R.S. and Kim, J.H. (2013) Production of Biallelic CMP-Neu5Ac
Hydroxylase Knock-Out Pigs. Scientific
Reports, 3, 1981-1990. http://dx.doi.org/10.1038/srep01981
Li, P., Estrada, J.L., Burlak, C. and Tector, A.J.
(2013) Biallelic Knockout of the Alpha-1, 3 Galactosyltransferase Gene in Porcine
Liver-Derived Cells Using Zinc Finger Nucleases. Journal of Surgical Research, 181, 39-45. http://dx.doi.org/10.1016/j.jss.2012.06.035
Toscano, M.G., Anderson, P., Munoz, P., Lucena, G., Cobo, M., Benabdellah,
K., Gregory, P.H., Holmes, M.C. and Martin, F. (2013) Use of Zinc-Finger Nucleases
to Knock out the WAS gene in K562 Cells: A Human Cellular Model for Wiskott-Aldrich
Syndrome. Disease Models Mechanisms, 6,
544-554. http://dx.doi.org/10.1242/dmm.010652
Cao, S.Z., Yue, C.H,
Li, X.R., Feng, C., Long, C. and Pan, D. K. (2013) Production of Myostatin Gene
Knockout Wuzhishan Miniature Pig Fibroblasts with Zinc-Finger Nucleases. HEREDITAS, 35, 778-785. http://dx.doi.org/10.3724/SP.J.1005.2013.00778