title: Comparative Analysis of piggyBac, CRISPR/Cas9 and TALEN Mediated BAC Transgenesis in the Zygote for the Generation of Humanized SIRPA Rats.
abstract: BAC transgenic mammalian systems offer an important platform for recapitulating human gene expression and disease modeling. While the larger body mass, and greater genetic and physiologic similarity to humans render rats well suited for reproducing human immune diseases and evaluating therapeutic strategies, difficulties of generating BAC transgenic rats have hindered progress. Thus, an efficient method for BAC transgenesis in rats would be valuable. Immunodeficient mice carrying a human SIRPA transgene have previously been shown to support improved human cell hematopoiesis. Here, we have generated for the first time, human SIRPA BAC transgenic rats, for which the gene is faithfully expressed, functionally active, and germline transmissible. To do this, human SIRPA BAC was modified with elements to work in coordination with genome engineering technologies-piggyBac, CRISPR/Cas9 or TALEN. Our findings show that piggyBac transposition is a more efficient approach than the classical BAC transgenesis, resulting in complete BAC integration with predictable end sequences, thereby permitting precise assessment of the integration site. Neither CRISPR/Cas9 nor TALEN increased BAC transgenesis. Therefore, an efficient generation of human SIRPA transgenic rats using piggyBac opens opportunities for expansion of humanized transgenic rat models in the future to advance biomedical research and therapeutic applications.
authors.affiliation: Center for Genetics, Children's Hospital Oakland Research Institute, CA 94609, Oakland, USA.
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Jung
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Ménoret
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Lucas
authors.initials: L
authors.lastname: Brusselle
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Tesson
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Usal
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Chenouard
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Remy
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Laure-Hélène
authors.initials: LH
authors.lastname: Ouisse
authors.affiliation: INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.|OSE Immunotherapeutics, 44000 Nantes, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Poirier
authors.affiliation: INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.|OSE Immunotherapeutics, 44000 Nantes, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Vanhove
authors.affiliation: Center for Genetics, Children's Hospital Oakland Research Institute, CA 94609, Oakland, USA.
authors.forename: Pieter J
authors.initials: PJ
authors.lastname: de Jong
authors.affiliation: Platform Rat Transgenesis Immunophenomic, SFR Francois Bonamy, CNRS UMS3556 Nantes, F44093, France.|INSERM UMR 1064-ITUN; CHU de Nantes, Nantes F44093, France.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Anegon
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D064113:CRISPR-Cas Systems; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D051381:Rats; D055647:Rats, Transgenic; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics; D019076:Transgenes; D015053:Zygote
chemical_list: D000943:Antigens, Differentiation; D011971:Receptors, Immunologic; C501920:SIRPA protein, human