TALEN-Mediated Modification of the Bovine Genome for Large-Scale Production of Human Serum Albumin

As an initial step towards creating genetically modified cattle as a biopharming source of recombinant human serum albumin (rHSA), we report modification of the bovine albumin (bA) locus by transcription activator-like effector nuclease (TALEN)-stimulated homology-directed repair (HDR). Pedigreed bovine fibroblasts were co-transfected with TALENs and an 11.5-kb human serum albumin (HSA) minigene donor construct, designed to simultaneously disrupt and replace bovine serum albumin (BSA) expression with controlled rHSA expression in both the liver and the milk. Targeted integration of the HSA minigene was confirmed in transfected fibroblasts at a frequency of approximately 11% and transgenic bovine embryos were produced from targeted fibroblasts using somatic cell nuclear transfer (SCNT). The research delineated here lays the foundation for the future generation of transgenic rHSA cattle with the potential to provide a large-scale, reliable, and quality-controlled source of rHSA.

[1]  Daichang Yang,et al.  Human serum albumin from recombinant DNA technology: challenges and strategies. , 2013, Biochimica et biophysica acta.

[2]  Daniel F. Voytas,et al.  Efficient TALEN-mediated gene knockout in livestock , 2012, Proceedings of the National Academy of Sciences.

[3]  Ning Li,et al.  Highly efficient modification of beta-lactoglobulin (BLG) gene via zinc-finger nucleases in cattle , 2011, Cell Research.

[4]  Lang Shen,et al.  Large-scale production of functional human serum albumin from transgenic rice seeds , 2011, Proceedings of the National Academy of Sciences.

[5]  A. Feinberg,et al.  Move over ZFNs , 2011, Nature Biotechnology.

[6]  J. Keith Joung,et al.  Targeted gene disruption in somatic zebrafish cells using engineered TALENs , 2011, Nature Biotechnology.

[7]  Yolanda Santiago,et al.  Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases , 2011, Proceedings of the National Academy of Sciences.

[8]  Thuy D. Vo,et al.  Transient cold shock enhances zinc-finger nuclease–mediated gene disruption , 2010, Nature Methods.

[9]  M. Destrempes,et al.  Production of recombinant albumin by a herd of cloned transgenic cattle , 2009, Transgenic Research.

[10]  Toni Cathomen,et al.  Zinc-finger Nucleases: The Next Generation Emerges. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[11]  M. Neville,et al.  Albumin transcytosis across the epithelium of the lactating mouse mammary gland , 2004, The Journal of physiology.

[12]  David Baltimore,et al.  Chimeric Nucleases Stimulate Gene Targeting in Human Cells , 2003, Science.

[13]  A. Schnieke,et al.  Production of gene-targeted sheep by nuclear transfer from cultured somatic cells , 2000, Nature.

[14]  D. Gardner,et al.  Enhanced rates of cleavage and development for sheep zygotes cultured to the blastocyst stage in vitro in the absence of serum and somatic cells: amino acids, vitamins, and culturing embryos in groups stimulate development. , 1994, Biology of reproduction.

[15]  M. Schechter,et al.  HIV transmission to patients with hemophilia by heat-treated, donor-screened factor concentrate. , 1990, CMAJ : Canadian Medical Association journal = journal de l'Association medicale canadienne.

[16]  J. Vilotte,et al.  Efficient tissue-specific expression of bovine α-lactalbumin in transgenic mice , 1989 .

[17]  J. Vilotte,et al.  Efficient tissue-specific expression of bovine alpha-lactalbumin in transgenic mice. , 1989, European Journal of Biochemistry.

[18]  R. Grappin,et al.  Milk protein analysis , 1989 .