title: Mutation of the XIST gene upregulates expression of X-linked genes but decreases the developmental rates of cloned male porcine embryos.
abstract: XIST is an X-linked, non-coding gene responsible for the cis induction of X-chromosome inactivation (XCI). Knockout of the XIST allele on an active X chromosome abolishes erroneous XCI and enhances the in vivo development of cloned mouse embryos by more than 10-fold. This study aimed to investigate whether a similar manipulation would improve cloning efficiency in pigs. A male, porcine kidney cell line containing an EGFP insert in exon 1 of the XIST gene, resulting in a knockout allele (XIST-KO), was generated by homologous recombination using transcription activator-like effector nucleases (TALENs). The expression of X-linked genes in embryos cloned from the XIST-KO kidney cells was significantly higher than in male embryos cloned from wild-type (WT) kidney cells, but remained lower than that of in vivo fertilization-produced counterparts. The XIST-KO cloned embryos also had a significantly lower blastocyst rate and a reduced full-term development rate compared to cloned WT embryos. These data suggested that while mutation of a XIST gene can partially rescue abnormal XCI, it cannot improve the developmental efficiency of cloned male porcine embryos-a deficiency that may be caused by incomplete rescue of abnormal XCI and/or by long-term drug selection of the XIST-KO nuclear donor cells, which might adversely affect the developmental efficiency of embryos created from them.
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Yang
authors.identifier: http://orcid.org/0000-0001-5560-052X
authors.initials: Y
authors.lastname: Yang
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Wu
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Dewu
authors.initials: D
authors.lastname: Liu
authors.affiliation: Guangdong Wen's Foodstuff Group Ltd., Yunfu, Guangdong, China.
authors.forename: Junsong
authors.initials: J
authors.lastname: Shi
authors.affiliation: Guangdong Wen's Foodstuff Group Ltd., Yunfu, Guangdong, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Zhou
authors.affiliation: Guangdong Wen's Foodstuff Group Ltd., Yunfu, Guangdong, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: He
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Jianping
authors.initials: J
authors.lastname: Quan
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Gengyuan
authors.initials: G
authors.lastname: Cai
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Enqin
authors.initials: E
authors.lastname: Zheng
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Zhenfang
authors.initials: Z
authors.lastname: Wu
authors.affiliation: National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.|Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Zicong
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D002460:Cell Line; D019976:Cloning, Organism; D050172:Genes, X-Linked; D008297:Male; D009154:Mutation; D062085:RNA, Long Noncoding / Q000096:biosynthesis / Q000235:genetics; D013552:Swine; D015854:Up-Regulation / Q000235:genetics*
chemical_list: D062085:RNA, Long Noncoding; C077885:XIST non-coding RNA
keywords: 
  XIST
  ; SCNT; XCI; cloning efficiency; pigs