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무균돼지 사육시설 계획을 위한 사례연구 : 공간구성 및 동선계획을 중심으로
권순정,성제경,염수청 한국의료복지시설학회 2006 의료·복지 건축 Vol.12 No.1
According to the increase of demand for human organs such as kidney, heart, pancreas, joint, and cornea for therapeutic transplantation, the production of alternative organs based on Gnotobiotic Pigs gains a lot of concerns all over the world. However, it is not common to design and build Gnotobiotic Pigs' facility, and there are only a few those facilities and planning principles for them. Considering the situation above, this paper tries to develop planning guidelines for space organization and circulation system of standardized Germ Free Pig's facility on the bases of case analysis. The results of this study are as follows. At first, four swine farms including a Gntobiotic Pig's facility has been analysed from the point of space organization and circulation system. Secondly, the space zoning of Gnotobiotic Pigs' facility has been proposed into 5 groups : pigs' area, adminstration area, operating room and laboratory, service area, and mechanical area. Space components of each group have been presented also. Finally, circulation system of Gnotobiotic Pigs' facility has been explored from a operational point of view. This, also, includes human circulation, pig's circulation, and goods' circulation. This study has some limitations because it does not consider the SOPs(standard operational policies) of that facility to the fullest measure and does not suggest space area of each part, either. Despite of some weaknesses, it is expected that this study can give some useful guidelines for the design and planning of Germ Free Pigs' facilities.
Yeom, Su-Cheong,Koo, Ok Jae,Park, Chung-Gyu,Lee, Byeong-Chun,Lee, Wang-Jae The Korean Society of Animal Reproduction 2013 Reproductive & developmental biology Vol.37 No.1
In order to investigate genetic stability and gene expression profile after cloning procedure, two groups of cloned pigs were used for swine leukocyte antigen (SLA) gene nucleotide alteration and microarray analyses. Each group was consist of cloned pigs derived from same cell line (n=3 and 4, respectively). Six SLA loci were analyzed for cDNA sequences and protein translations. In total, 16 SLA alleles were identified and there were no evidence of SLA nucleotide alteration. All SLA sequences and protein translations were identical among the each pig in the same group. On the other hand, microarray assay was performed for profiling gene expression of the cloned pigs. In total, 43,603 genes were analyzed and 2,150~4,300 reliably hybridized spots on the each chip were selected for further analysis. Even though the cloned pigs in the same group had identical genetic background, 18.6~47.3% of analyzed genes were differentially expressed in between each cloned pigs. Furthermore, on gene clustering analysis, some cloned pigs showed abnormal physiological phenotypes such as inflammation, cancer or cardiomyopathy. We assumed that individual environmental adaption, sociality and rank in the pen might have induced these different phenotypes. In conclusion, the results of the present study indicate that SLA locus genes appear to be stable following SCNT. However, gene expressions and phenotypes between cloned pigs derived from the same cell line were not identical even under the same rearing conditions.
YEOM, Su-Cheong,YU, Sun-A,CHOI, Eun-Young,LEE, Byeong-Chun,LEE, Wang-Jae Japanese Association for Laboratory Animal Science 2009 Experimental animals Vol.58 No.5
<P>We investigated the prevalence of <I>Helicobacter hepaticus</I>, murine norovirus (MNV), and <I>Pneumocystis carinii</I> and the efficacy of cross-fostering for their eradication in 49 genetically engineered mouse (GEM) strains at our institute. Prevalences of <I>H. hepaticus</I>, MNV, and <I>P. carinii</I> were 33.9, 36.5, and 8.6%, respectively, and immunodeficient strains showed relatively higher prevalence of the 3 pathogens than immunocompetent strains. Additionally, the same immune phenotype strains showed similar prevalences. Furthermore, it was found that NKT cells might play a role in <I>H. hepaticus</I> resistance. Interestingly, there was a high incidence of <I>H. hepaticus</I> and MNV multiple infection. Strains with single or multiple infections of <I>H. hepaticus</I>, MNV, and/or <I>P. carinii</I> were selected, and cross-fostering was conducted. Cross-fosterings were successful at eradicating <I>P. carinii</I>, but there were some failures for <I>H. hepaticus</I> and MNV, and the efficacy of eradication was relatively low compared with previous studies. We thought that this low efficacy might have been due to persistent infection and the high suscepibility to <I>H. hepaticus</I> and MNV of immunodeficient GEM strains. Therefore, cross-fostering may be appropriate for <I>P. carinii</I> eradication, but be inappropriate for repopulation of a new breeding colony with <I>H. hepaticus</I> or MNV infected GEM strains. Our findings provide basic data on maintenance, strain susceptibility, and successful rederivation, especially for GEMs.</P>
CRISPR and SSTR mediated knock-in with mouse embryos
Su-Cheong Yeom 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11
The CRISPR/Cas9 system is widely applied in genome engineering due to its simplicity and versatility. Although this has revolutionized genome-editing technology, knock-in animal generation via homology directed repair (HDR) is not as efficient as non-homologous end-joining DNA-repair-dependent knockout. Although its double-strand break activity may vary, Cas9 derived from Streptococcus pyogenens allows robust design of single-guide RNAs (sgRNAs) within the target sequence; However, prescreening for different sgRNA activities delays the process of transgenic animal generation. To overcome this limitation, multiple sets of different sgRNAs were examined for their knock-in efficiency. We discovered profound advantages associated with single-stranded oligo-donor-mediated HDR processes using overlapping sgRNAs (sharing at least five base pairs of the target sites) as compared with using non-overlapping sgRNAs for knock-in mouse generation. Studies utilizing cell lines revealed shorter sequence deletions near target mutations using overlapping sgRNAs as compared with those observed using non-overlapping sgRNAs, which may favor the HDR process. Using this simple method, we successfully generated several transgenic mouse lines harboring loxP insertions or single-nucleotide substitutions with a highly efficiency of 18~38%. Our results demonstrate a simple and efficient method for generating transgenic animals harboring foreign-sequence knock-ins or short-nucleotide substitutions by the use of overlapping sgRNAs.
YEOM, Su-Cheong,PARK, Chung-Gyu,LEE, Byeong-Chun,LEE, Wang-Jae Blackwell Publishing Asia 2010 ANIMAL SCIENCE JOURNAL Vol.81 No.2
<P>ABSTRACT</P><P>Seoul National University (SNU) miniature pigs represent a closed colony with 24 founder pigs and a well preserved pedigree. Characterization using mRNA sequence analysis was conducted for 6 swine leukocyte antigen (SLA) loci in parental or founder pigs, and 17 defined alleles were detected. Based on these complete coding sequences, 17 sequence specific primers (SSPs) were designed for polymorphic sites. To validate the specificity of each allele SSP, the PCR-SSP was conducted with defined allele clones as templates. PCR-SSP was conducted with the hot start polymerase and touch-down PCR. The parental or found SNU miniature pigs showed overall SLA class I and II heterozygotes. Using the established PCR-SSP method, we conducted SLA typing for breeding stock including 2 pedigreed pigs and identified the novel SLA class II homozygote haplotye (<I>DRA</I>*<I>0201, DRB1</I>*<I>0403, DQA</I>*<I>0102</I> and <I>DQB1</I>*<I>0701</I>) and 2 SLA homozygote pig lines: SLA class I Hp-3.0 and class II Hp-0.3, and SLA class I Hp-2.0 and class II Hp-0.2. We thought that our PCR-SSP SLA typing method could be applicable for new SLA homozygote line establishment by assignment and scheduled breeding.</P>
YEOM, Su-Cheong,KONG, Dae-Young,PARK, Chung-Gyu,LEE, Byeong-Chun,LEE, Wang-Jae Blackwell Publishing Asia 2010 ANIMAL SCIENCE JOURNAL Vol.81 No.2
<P>ABSTRACT</P><P>Seoul National University (SNU) miniature pigs are originated from the Minnesota miniature pig. This study was conducted to investigate the maternal origin of SNU (Minnesota) miniature pigs and their phylogenetic relationships by analyzing the mitochondrial DNA (mtDNA) <I>D-loop</I> (control region) sequence. Two mtDNA <I>D-loop</I> sequences of the SNU miniature pigs were identified. On an unweighted pair-group method with an arithmetic mean (UPGMA) phylogenetic tree analysis, the large white was the pig breed closest to the SNU miniature pig, and the pairwise distance analysis showed the same result. While mtDNA sequences of 4 pig breeds which were used to establish Minnesota miniature pig were not known, our result might be different from the history of the Minnesota miniature pig development. In conclusion, we thought that some haplotypes of the Minnesota miniature pig maternally were originated from the Large white pig, or that wild pigs had similar mtDNA sequences to the Large white pig, and all SNU miniature pigs were derived from this colony.</P>