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Gene knockout and knockin by zinc-finger nucleases: current status and perspectives

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Abstract

Zinc-finger nucleases (ZFNs) are engineered site-specific DNA cleavage enzymes that may be designed to recognize long target sites and thus cut DNA with high specificity. ZFNs mediate permanent and targeted genetic alteration via induction of a double-strand break at a specific genomic site. Compared to conventional homology-based gene targeting, ZFNs can increase the targeting rate by up to 100,000-fold; gene disruption via mutagenic DNA repair is similarly efficient. The utility of ZFNs has been shown in many organisms, including insects, amphibians, plants, nematodes, and several mammals, including humans. This broad range of tractable species renders ZFNs a useful tool for improving the understanding of complex physiological systems, to produce transgenic animals, cell lines, and plants, and to treat human disease.

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Abbreviations

BKG:

Beta-lactoglobulin

CCR5 :

Human chemokine receptors 5 gene

CHO:

Chinese hamster ovary

Dat :

Dopamine transporter

DHFR :

Dihydrofolate reductase

DSB:

Double-strand break

ES cells:

Embryonic stem cells

FACS:

Fluorescence-activated cell sorting

FUT8 :

α-1,6-fucosyltransferase

GFP:

Green fluorescence protein

GGTA1 :

α1,3-galactosyltransferase (Gal)

GPI-AP:

Glycosyl-phosphatidyl-inositol anchored proteins

GS:

Glutamine synthetase

HDR:

Homology-direct repair

hFIX:

Human factor IX

hif1 α :

Hypoxia-inducible factor 1α

HR:

Homologous recombination

HSPCs:

Hematopoietic stem progenitor cells

IDLV:

Integrase-defective lentivirus

IgM:

Immunoglobulin M

IL2Rγ:

Interleukin-2 receptor common γ-chain

iPS cells:

Induced pluripotent stem cells

Jag1 :

Jagged 1

KO:

Knockout

KI:

Knockin

Mdr1a :

Multidrug-resistant 1a

NHEJ:

Non-homologous end joining

Notch3 :

Notch homolog 3

OPEN:

Oligomerized pool engineering

ORF:

Open reading frame

OTS:

Off-target site

Ppar-γ:

Peroxisome proliferator-activated receptor-γ

SCID:

Severe combined immune deficiency

SCNT:

Somatic cell nuclear transfer

tfr2 :

Telomerase, transferrin receptor 2

WT:

Wild type

ZF:

Zinc-finger

ZFN:

Zinc-finger nucleases

ZFP:

Zinc-finger proteins

References

  1. Rouet P, Smih F, Jasin M (1994) Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells. Proc Natl Acad Sci USA 91(13):6064–6068

    Article  PubMed  CAS  Google Scholar 

  2. Miller J, McLachlan AD, Klug A (1985) Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes. EMBO J 4(6):1609–1614

    PubMed  CAS  Google Scholar 

  3. Pabo CO, Peisach E, Grant RA (2001) Design and selection of novel Cys2His2 zinc finger proteins. Annu Rev Biochem 70:313–340

    Article  PubMed  CAS  Google Scholar 

  4. Pavletich NP, Pabo CO (1991) Zinc finger-DNA recognition: crystal structure of a Zif268-DNA complex at 2.1 A. Science 252(5007):809–817

    Article  PubMed  CAS  Google Scholar 

  5. Kim YG, Cha J, Chandrasegaran S (1996) Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain. Proc Natl Acad Sci USA 93(3):1156–1160

    Article  PubMed  CAS  Google Scholar 

  6. Smith J, Bibikova M, Whitby FG, Reddy AR, Chandrasegaran S, Carroll D (2000) Requirements for double-strand cleavage by chimeric restriction enzymes with zinc finger DNA-recognition domains. Nucleic Acids Res 28(17):3361–3369

    Article  PubMed  CAS  Google Scholar 

  7. Bibikova M, Carroll D, Segal DJ, Trautman JK, Smith J, Kim YG, Chandrasegaran S (2001) Stimulation of homologous recombination through targeted cleavage by chimeric nucleases. Mol Cell Biol 21(1):289–297

    Article  PubMed  CAS  Google Scholar 

  8. Bibikova M, Golic M, Golic KG, Carroll D (2002) Targeted chromosomal cleavage and mutagenesis in Drosophila using zinc-finger nucleases. Genetics 161(3):1169–1175

    PubMed  CAS  Google Scholar 

  9. Doyon Y, Choi VM, Xia DF, Vo TD, Gregory PD, Holmes MC (2010) Transient cold shock enhances zinc-finger nuclease-mediated gene disruption. Nat Methods 7(6):459–460

    Article  PubMed  CAS  Google Scholar 

  10. Roobol A, Carden MJ, Newsam RJ, Smales CM (2009) Biochemical insights into the mechanisms central to the response of mammalian cells to cold stress and subsequent rewarming. FEBS J 276(1):286–302

    Article  PubMed  CAS  Google Scholar 

  11. Qiu P, Shandilya H, D’Alessio JM, O’Connor K, Durocher J, Gerard GF (2004) Mutation detection using surveyor nuclease. Biotechniques 36(4):702–707

    PubMed  CAS  Google Scholar 

  12. Kim S, Lee HJ, Kim E, Kim JS (2010) Analysis of targeted chromosomal deletions induced by zinc finger nucleases. Cold Spring Harb Protoc 2010 (8):pdb prot5477. doi:10.1101/pdb.prot5477

  13. Santiago Y, Chan E, Liu PQ, Orlando S, Zhang L, Urnov FD, Holmes MC, Guschin D, Waite A, Miller JC, Rebar EJ, Gregory PD, Klug A, Collingwood TN (2008) Targeted gene knockout in mammalian cells by using engineered zinc-finger nucleases. Proc Natl Acad Sci USA 105(15):5809–5814

    Article  PubMed  CAS  Google Scholar 

  14. Liu PQ, Chan EM, Cost GJ, Zhang L, Wang JB, Miller JC, Guschin DY, Reik A, Holmes MC, Mott JE, Collingwood TN, Gregory PD (2010) Generation of a triple-gene knockout mammalian cell line using engineered zinc-finger nucleases. Biotechnol Bioeng 106(1):97–105

    PubMed  CAS  Google Scholar 

  15. Malphettes L, Freyvert Y, Chang J, Liu PQ, Chan E, Miller JC, Zhou Z, Nguyen T, Tsai C, Snowden AW, Collingwood TN, Gregory PD, Cost GJ (2010) Highly efficient deletion of FUT8 in CHO cell lines using zinc-finger nucleases yields cells that produce completely nonfucosylated antibodies. Biotechnol Bioeng 106(5):774–783

    Article  PubMed  CAS  Google Scholar 

  16. Yu S, Luo J, Song Z, Ding F, Dai Y, Li N (2011) Highly efficient modification of beta-lactoglobulin (BLG) gene via zinc-finger nucleases in cattle. Cell Res 21(11):1638–1640

    Article  PubMed  CAS  Google Scholar 

  17. Whyte JJ, Zhao J, Wells KD, Samuel MS, Whitworth KM, Walters EM, Laughlin MH, Prather RS (2011) Gene targeting with zinc finger nucleases to produce cloned eGFP knockout pigs. Mol Reprod Dev 78(1):2

    Article  PubMed  CAS  Google Scholar 

  18. Hauschild J, Petersen B, Santiago Y, Queisser AL, Carnwath JW, Lucas-Hahn A, Zhang L, Meng X, Gregory PD, Schwinzer R, Cost GJ, Niemann H (2011) Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases. Proc Natl Acad Sci USA 108(29):12013–12017

    Article  PubMed  CAS  Google Scholar 

  19. Gandhi M, Evdokimova VN, TC K, Nikiforova MN, Kelly LM, Stringer JR, Bakkenist CJ, Nikiforov YE (2012) Homologous chromosomes make contact at the sites of double-strand breaks in genes in somatic G0/G1-phase human cells. Proc Natl Acad Sci USA 109(24):9454–9459

    Article  PubMed  CAS  Google Scholar 

  20. Moehle EA, Rock JM, Lee YL, Jouvenot Y, DeKelver RC, Gregory PD, Urnov FD, Holmes MC (2007) Targeted gene addition into a specified location in the human genome using designed zinc finger nucleases. Proc Natl Acad Sci USA 104(9):3055–3060

    Article  PubMed  CAS  Google Scholar 

  21. Deng C, Capecchi MR (1992) Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus. Mol Cell Biol 12(8):3365–3371

    PubMed  CAS  Google Scholar 

  22. Orlando SJ, Santiago Y, DeKelver RC, Freyvert Y, Boydston EA, Moehle EA, Choi VM, Gopalan SM, Lou JF, Li J, Miller JC, Holmes MC, Gregory PD, Urnov FD, Cost GJ (2010) Zinc-finger nuclease-driven targeted integration into mammalian genomes using donors with limited chromosomal homology. Nucleic Acids Res 38(15):e152. doi:10.1093/nar/gkq512

    Article  PubMed  Google Scholar 

  23. Pruett-Miller SM, Connelly JP, Maeder ML, Joung JK, Porteus MH (2008) Comparison of zinc finger nucleases for use in gene targeting in mammalian cells. Mol Ther 16(4):707–717

    Article  PubMed  CAS  Google Scholar 

  24. Isalan M, Choo Y, Klug A (1997) Synergy between adjacent zinc fingers in sequence-specific DNA recognition. Proc Natl Acad Sci USA 94(11):5617–5621

    Article  PubMed  CAS  Google Scholar 

  25. Ramirez CL, Foley JE, Wright DA, Muller-Lerch F, Rahman SH, Cornu TI, Winfrey RJ, Sander JD, Fu F, Townsend JA, Cathomen T, Voytas DF, Joung JK (2008) Unexpected failure rates for modular assembly of engineered zinc fingers. Nat Methods 5(5):374–375

    Article  PubMed  CAS  Google Scholar 

  26. Sander JD, Dahlborg EJ, Goodwin MJ, Cade L, Zhang F, Cifuentes D, Curtin SJ, Blackburn JS, Thibodeau-Beganny S, Qi Y, Pierick CJ, Hoffman E, Maeder ML, Khayter C, Reyon D, Dobbs D, Langenau DM, Stupar RM, Giraldez AJ, Voytas DF, Peterson RT, Yeh JR, Joung JK (2011) Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA). Nat Methods 8(1):67–69

    Article  PubMed  CAS  Google Scholar 

  27. Maeder ML, Thibodeau-Beganny S, Osiak A, Wright DA, Anthony RM, Eichtinger M, Jiang T, Foley JE, Winfrey RJ, Townsend JA, Unger-Wallace E, Sander JD, Muller-Lerch F, Fu F, Pearlberg J, Gobel C, Dassie JP, Pruett-Miller SM, Porteus MH, Sgroi DC, Iafrate AJ, Dobbs D, McCray PB Jr, Cathomen T, Voytas DF, Joung JK (2008) Rapid “open-source” engineering of customized zinc-finger nucleases for highly efficient gene modification. Mol Cell 31(2):294–301

    Article  PubMed  CAS  Google Scholar 

  28. Reyon D, Kirkpatrick JR, Sander JD, Zhang F, Voytas DF, Joung JK, Dobbs D, Coffman CR (2011) ZFNGenome: a comprehensive resource for locating zinc finger nuclease target sites in model organisms. BMC Genomics 12:83–91

    Article  PubMed  CAS  Google Scholar 

  29. Doyon Y, McCammon JM, Miller JC, Faraji F, Ngo C, Katibah GE, Amora R, Hocking TD, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Amacher SL (2008) Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases. Nat Biotechnol 26(6):702–708

    Article  PubMed  CAS  Google Scholar 

  30. Geurts AM, Cost GJ, Freyvert Y, Zeitler B, Miller JC, Choi VM, Jenkins SS, Wood A, Cui X, Meng X, Vincent A, Lam S, Michalkiewicz M, Schilling R, Foeckler J, Kalloway S, Weiler H, Menoret S, Anegon I, Davis GD, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Jacob HJ, Buelow R (2009) Knockout rats via embryo microinjection of zinc-finger nucleases. Science 325(5939):433

    Article  PubMed  CAS  Google Scholar 

  31. Perez EE, Wang J, Miller JC, Jouvenot Y, Kim KA, Liu O, Wang N, Lee G, Bartsevich VV, Lee YL, Guschin DY, Rupniewski I, Waite AJ, Carpenito C, Carroll RG, Orange JS, Urnov FD, Rebar EJ, Ando D, Gregory PD, Riley JL, Holmes MC, June CH (2008) Establishment of HIV-1 resistance in CD4+T cells by genome editing using zinc-finger nucleases. Nat Biotechnol 26(7):808–816

    Article  PubMed  CAS  Google Scholar 

  32. Meng X, Noyes MB, Zhu LJ, Lawson ND, Wolfe SA (2008) Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases. Nat Biotechnol 26(6):695–701

    Article  PubMed  CAS  Google Scholar 

  33. Miller JC, Holmes MC, Wang J, Guschin DY, Lee YL, Rupniewski I, Beausejour CM, Waite AJ, Wang NS, Kim KA, Gregory PD, Pabo CO, Rebar EJ (2007) An improved zinc-finger nuclease architecture for highly specific genome editing. Nat Biotechnol 25(7):778–785

    Article  PubMed  CAS  Google Scholar 

  34. Szczepek M, Brondani V, Buchel J, Serrano L, Segal DJ, Cathomen T (2007) Structure-based redesign of the dimerization interface reduces the toxicity of zinc-finger nucleases. Nat Biotechnol 25(7):786–793

    Article  PubMed  CAS  Google Scholar 

  35. Sollu C, Pars K, Cornu TI, Thibodeau-Beganny S, Maeder ML, Joung JK, Heilbronn R, Cathomen T (2010) Autonomous zinc-finger nuclease pairs for targeted chromosomal deletion. Nucleic Acids Res 38(22):8269–8276

    Article  PubMed  Google Scholar 

  36. Doyon Y, Vo TD, Mendel MC, Greenberg SG, Wang J, Xia DF, Miller JC, Urnov FD, Gregory PD, Holmes MC (2011) Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures. Nat Methods 8(1):74–79

    Article  PubMed  CAS  Google Scholar 

  37. Yang D, Yang H, Li W, Zhao B, Ouyang Z, Liu Z, Zhao Y, Fan N, Song J, Tian J, Li F, Zhang J, Chang L, Pei D, Chen YE, Lai L (2011) Generation of PPARgamma mono-allelic knockout pigs via zinc-finger nucleases and nuclear transfer cloning. Cell Res 21(6):979–982

    Article  PubMed  CAS  Google Scholar 

  38. Yusa K, Rashid ST, Strick-Marchand H, Varela I, Liu PQ, Paschon DE, Miranda E, Ordonez A, Hannan NR, Rouhani FJ, Darche S, Alexander G, Marciniak SJ, Fusaki N, Hasegawa M, Holmes MC, Di Santo JP, Lomas DA, Bradley A, Vallier L (2011) Targeted gene correction of alpha1-antitrypsin deficiency in induced pluripotent stem cells. Nature 478(7369):391–394

    Article  PubMed  CAS  Google Scholar 

  39. Urnov FD, Miller JC, Lee YL, Beausejour CM, Rock JM, Augustus S, Jamieson AC, Porteus MH, Gregory PD, Holmes MC (2005) Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature 435(7042):646–651

    Article  PubMed  CAS  Google Scholar 

  40. Holt N, Wang J, Kim K, Friedman G, Wang X, Taupin V, Crooks GM, Kohn DB, Gregory PD, Holmes MC, Cannon PM (2010) Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo. Nat Biotechnol 28(8):839–847

    Article  PubMed  CAS  Google Scholar 

  41. Bobis-Wozowicz S, Osiak A, Rahman SH, Cathomen T (2011) Targeted genome editing in pluripotent stem cells using zinc-finger nucleases. Methods 53(4):339–346

    Article  PubMed  CAS  Google Scholar 

  42. Connelly JP, Barker JC, Pruett-Miller S, Porteus MH (2010) Gene correction by homologous recombination with zinc finger nucleases in primary cells from a mouse model of a generic recessive genetic disease. Mol Ther 18(6):1103–1110

    Article  PubMed  CAS  Google Scholar 

  43. Hockemeyer D, Soldner F, Beard C, Gao Q, Mitalipova M, DeKelver RC, Katibah GE, Amora R, Boydston EA, Zeitler B, Meng X, Miller JC, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Jaenisch R (2009) Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27(9):851–857

    Article  PubMed  CAS  Google Scholar 

  44. Zou J, Maeder ML, Mali P, Pruett-Miller SM, Thibodeau-Beganny S, Chou BK, Chen G, Ye Z, Park IH, Daley GQ, Porteus MH, Joung JK, Cheng L (2009) Gene targeting of a disease-related gene in human induced pluripotent stem and embryonic stem cells. Cell Stem Cell 5(1):97–110

    Article  PubMed  CAS  Google Scholar 

  45. Lombardo A, Genovese P, Beausejour CM, Colleoni S, Lee YL, Kim KA, Ando D, Urnov FD, Galli C, Gregory PD, Holmes MC, Naldini L (2007) Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery. Nat Biotechnol 25(11):1298–1306

    Article  PubMed  CAS  Google Scholar 

  46. Cost GJ, Freyvert Y, Vafiadis A, Santiago Y, Miller JC, Rebar E, Collingwood TN, Snowden A, Gregory PD (2010) BAK and BAX deletion using zinc-finger nucleases yields apoptosis-resistant CHO cells. Biotechnol Bioeng 105(2):330–340

    Article  PubMed  CAS  Google Scholar 

  47. Young JJ, Cherone JM, Doyon Y, Ankoudinova I, Faraji FM, Lee AH, Ngo C, Guschin DY, Paschon DE, Miller JC, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Harland RM, Zeitler B (2011) Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases. Proc Natl Acad Sci USA 108(17):7052–7057

    Article  PubMed  CAS  Google Scholar 

  48. Foley JE, Yeh JR, Maeder ML, Reyon D, Sander JD, Peterson RT, Joung JK (2009) Rapid mutation of endogenous zebrafish genes using zinc finger nucleases made by oligomerized pool engineering (OPEN). PLoS One 4(2):e4348. doi:10.1371/journal.pone.0004348

    Article  PubMed  Google Scholar 

  49. Wood AJ, Lo TW, Zeitler B, Pickle CS, Ralston EJ, Lee AH, Amora R, Miller JC, Leung E, Meng X, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Meyer BJ (2011) Targeted genome editing across species using ZFNs and TALENs. Science 333(6040):307

    Article  PubMed  CAS  Google Scholar 

  50. Carbery ID, Ji D, Harrington A, Brown V, Weinstein EJ, Liaw L, Cui X (2010) Targeted genome modification in mice using zinc-finger nucleases. Genetics 186(2):451–459

    Article  PubMed  CAS  Google Scholar 

  51. Meyer M, de Angelis MH, Wurst W, Kuhn R (2010) Gene targeting by homologous recombination in mouse zygotes mediated by zinc-finger nucleases. Proc Natl Acad Sci USA 107(34):15022–15026

    Article  PubMed  CAS  Google Scholar 

  52. Cui X, Ji D, Fisher DA, Wu Y, Briner DM, Weinstein EJ (2011) Targeted integration in rat and mouse embryos with zinc-finger nucleases. Nat Biotechnol 29(1):64–67

    Article  PubMed  CAS  Google Scholar 

  53. Uren AG, Kool J, Berns A, van Lohuizen M (2005) Retroviral insertional mutagenesis: past, present and future. Oncogene 24(52):7656–7672

    Article  PubMed  CAS  Google Scholar 

  54. Perez-Pinera P, Ousterout DG, Brown MT, Gersbach CA (2011) Gene targeting to the ROSA26 locus directed by engineered zinc finger nucleases. Nucleic Acids Res 40(8):3741–3752

    Article  PubMed  Google Scholar 

  55. Li H, Haurigot V, Doyon Y, Li T, Wong SY, Bhagwat AS, Malani N, Anguela XM, Sharma R, Ivanciu L, Murphy SL, Finn JD, Khazi FR, Zhou S, Paschon DE, Rebar EJ, Bushman FD, Gregory PD, Holmes MC, High KA (2011) In vivo genome editing restores haemostasis in a mouse model of haemophilia. Nature 475(7355):217–221

    Article  PubMed  CAS  Google Scholar 

  56. Remy S, Tesson L, Menoret S, Usal C, Scharenberg AM, Anegon I (2010) Zinc-finger nucleases: a powerful tool for genetic engineering of animals. Transgenic Res 19(3):363–371

    Article  PubMed  CAS  Google Scholar 

  57. Mashimo T, Takizawa A, Voigt B, Yoshimi K, Hiai H, Kuramoto T, Serikawa T (2010) Generation of knockout rats with X-linked severe combined immunodeficiency (X-SCID) using zinc-finger nucleases. PLoS One 5(1):e8870. doi:10.1371/journal.pone.0008870

    Article  PubMed  Google Scholar 

  58. Flisikowska T, Thorey IS, Offner S, Ros F, Lifke V, Zeitler B, Rottmann O, Vincent A, Zhang L, Jenkins S, Niersbach H, Kind AJ, Gregory PD, Schnieke AE, Platzer J (2011) Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases. PLoS One 6(6):e21045. doi:10.1371/journal.pone.0021045

    Article  PubMed  CAS  Google Scholar 

  59. Rogers CS, Stoltz DA, Meyerholz DK, Ostedgaard LS, Rokhlina T, Taft PJ, Rogan MP, Pezzulo AA, Karp PH, Itani OA, Kabel AC, Wohlford-Lenane CL, Davis GJ, Hanfland RA, Smith TL, Samuel M, Wax D, Murphy CN, Rieke A, Whitworth K, Uc A, Starner TD, Brogden KA, Shilyansky J, McCray PB Jr, Zabner J, Prather RS, Welsh MJ (2008) Disruption of the CFTR gene produces a model of cystic fibrosis in newborn pigs. Science 321(5897):1837–1841

    Article  PubMed  CAS  Google Scholar 

  60. van den Heuvel M, Sorop O, Koopmans SJ, Dekker R, de Vries R, van Beusekom HM, Eringa EC, Duncker DJ, Danser AH, van der Giessen WJ (2012) Coronary microvascular dysfunction in a porcine model of early atherosclerosis and diabetes. Am J Physiol Heart Circ Physiol 302(1):H85–H94

    Article  PubMed  Google Scholar 

  61. Mikkelsen M, Moller A, Jensen LH, Pedersen A, Harajehi JB, Pakkenberg H (1999) MPTP-induced Parkinsonism in minipigs: a behavioral, biochemical, and histological study. Neurotoxicol Teratol 21(2):169–175

    Article  PubMed  CAS  Google Scholar 

  62. Cooper DK, Ayares D (2011) The immense potential of xenotransplantation in surgery. Int J Surg 9(2):122–129

    Article  PubMed  Google Scholar 

  63. Petersen B, Lucas-Hahn A, Oropeza M, Hornen N, Lemme E, Hassel P, Queisser AL, Niemann H (2008) Development and validation of a highly efficient protocol of porcine somatic cloning using preovulatory embryo transfer in peripubertal gilts. Cloning Stem Cells 10(3):355–362

    Article  PubMed  CAS  Google Scholar 

  64. Watanabe M, Umeyama K, Matsunari H, Takayanagi S, Haruyama E, Nakano K, Fujiwara T, Ikezawa Y, Nakauchi H, Nagashima H (2010) Knockout of exogenous EGFP gene in porcine somatic cells using zinc-finger nucleases. Biochem Biophys Res Commun 402(1):14–18

    Article  PubMed  CAS  Google Scholar 

  65. Whyte JJ, Prather RS (2011) Zinc finger nucleases to create custom-designed modifications in the swine (Sus scrofa) genome. J Anim Sci 90(4):1111–1117

    Article  PubMed  Google Scholar 

  66. Li P, Estrada JL, Burlak C, Tector AJ (2012) Biallelic knockout of the alpha-1,3 galactosyltransferase gene in porcine liver-derived cells using zinc finger nucleases. J Surg Res. doi:10.1016/j.jss.2012.06.035

    Google Scholar 

  67. Chu X, Zhang Z, Yabut J, Horwitz S, Levorse J, Li XQ, Zhu L, Lederman H, Ortiga R, Strauss J, Li X, Owens KA, Dragovic J, Vogt T, Evers R, Shin MK (2012) Characterization of multidrug resistance 1a/P-glycoprotein knockout rats generated by zinc finger nucleases. Mol Pharmacol 81(2):220–227

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut, Mariensee, Hoeltystrasse 10, 31535, Neustadt a. Rbge., Germany

    J. Hauschild-Quintern, B. Petersen & H. Niemann

  2. Rebirth, Cluster of Excellence, Hannover Medical School, Hannover, Germany

    H. Niemann

  3. Sangamo BioSciences, 501 Canal Blvd., Richmond, CA, 94804, USA

    G. J. Cost

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  1. J. Hauschild-Quintern
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  2. B. Petersen
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Correspondence to H. Niemann.

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Hauschild-Quintern, J., Petersen, B., Cost, G.J. et al. Gene knockout and knockin by zinc-finger nucleases: current status and perspectives. Cell. Mol. Life Sci. 70, 2969–2983 (2013). https://doi.org/10.1007/s00018-012-1204-1

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