돼지의 체세포 핵이식(Somatic cell nuclear transfer,SCNT)은 인간에게 약리적 효과가 있는 단백질, 이종 간 장기이식(xenotransplantation)에 사용되는 장기, 질병 연 구 목적의 모델 동물을 제공한다. 특히 형질전환 돼지를 활용한 심장 이식이 세계 최초로 성공한 후 형질전환 돼 지 생산의 안정화는 다음 연구를 위한 중요한 점으로 대 두되고 있으나, 미니돼지의 체세포 핵이식 배아의 생산 효율은 아직 낮은 실정이다. 형질전환의 성공은 양질의 SCNT 배아 생산에서 시작되어야 한다. 이러한 SCNT 배 아의 생산 효율을 향상할 수 있는 요인 중에는 공여 세포 의 형태가 있으며, 성공적인 공여 세포의 생산을 위해서 는 종축에 따른 세포의 특성을 파악하여야 하고, 혈액형 의 차이에서 발생하는 문제점 해결을 위해 OO 타입의 선 별이 필요하다. 본 연구에서는 지속적인 계대 배양을 통 하여 공여 세포로 사용되는 미니돼지의 태아섬유아세포의 계대 배양 조건을 확립하고자 한다. 또한 미니돼지의 혈 액형을 PCR 기반으로 분석하여 분류하고 OO 타입의 선 별을 통하여 이종 간 이식에 용이하게 공여 세포의 조건 을 확립하였다. 이후 sgRNA(single guide RNA)를 사용하 여 CRISPR-Cpf1로 GGTA1(α-1,3 galactosyl-transferase) 유전자를 knock-out 한 미니돼지의 생산으로, 급성면역반 응을 유발하는 Gal(1,3)Gal epitope이 제거된 미니돼지의 세포 주를 구축 및 체세포 핵이식을 통해 GGTA1 knock-out 미니돼지를 생산하였으며, 이러한 연구는 이후 체세포 핵이식 및 이종 간 장기이식에 중요한 기초자료로 사용될 것이라고 생각된다.

Severe combined immune deficiency (SCID) pig is very important research model for biomedical research, such as the development of humanized tissues and organs for transplantation and long-term evaluation of transplanted cancer or stem cell of human origin. FOXN1 gene encodes a transcription factor essential for the development and function of thymic epithelial cells (TECs), the primary lymphoid organ that supports T-cell development and selection. In this study, we are going to produce the FOXN1 KO SCID pigs using the Crispr/Cpf1 method. Porcine genomic DNA sequences were analyzed and the target sequences were selected using a web tool, Benchling (https://benchling.com/). The designed crDNA oligos was synthesized by the Oligonucleotide Synthesis Service (Macrogen Inc., Seoul, Korea). To generate the AsCpf1-mCherry-Puro construct, pTE4396 (#74041; Addgene, Cambridge, MA, USA) was modified by removing the NeoR/KanR sequence using BstBI and SmaI. Then, the mCherry-Puro sequence from pSicoR-Ef1a-mCh-Puro (#31845; Addgene, Cambridge, MA, USA) digested with the same restriction enzymes was inserted into the aforementioned NeoR/KanR-deleted vector. The crDNA #1 or crDNA #2 was inserted into the pTE4396 and AsCpf1-mCherry-Puro vectors in the U6 promoter region using BsmBI enzyme, respectively. The two vectors were transfected with lipofectamine 3000 (Life Technologies, Grand Island, NY, USA) and selected with puromycin and G-418 antibiotics. As a result, we established a cell line into which two vectors (pTE4396+crFOXN1#2 and AsCpf1- mCherry-Puro+ crFOXN1#1) and were inserted. Further studies are needed to characterize FOXN1 KO cell lines.

Industrial emissions, mainly from industrial complexes, are important sources of ambient Volatile Organic Compounds (VOCs). Identification of the significant VOC sources from industrial complexes has practical significance for emission reduction. VOC samples were collected from July 2019 to June 2020. A Positive Matrix Factorization (PMF) receptor model was used to evaluate the VOC sources in the area. Four sources were identified by PMF analysis, including coating-1, coating-2, printing, and vehicle exhaust. The coating-1 source was revealed to have the highest contribution (41.5%), followed by coating-2 (23.9%), printing (23.1%), and vehicle exhaust (11.6%). The source showing the highest contribution was coating emissions, originating from the northwest to southwest of the sample site. It also relates to facilities that produce auto parts. The major components of VOC emissions from the coating facilities were toluene, m,p-xylene, ethylbenzene, o-xylene, and butyl acetate. Industrial emissions should be the top priority to meet the relevant control criteria, followed by vehicular emissions. This study provides a strategy for VOC source apportionment from an industrial complex, which is helpful in the development of targeted control strategies.