연구 - 유전자 전이방법을 이용한 효율적인 형질전환 닭 생산 기술 확립 - 인위적 계란 성분 조절이 가능한 새로운 형질의 닭 품종 대량 생산 체계 확립

한재용,박태섭,Han, Jae-Yong,Park, Tae-Seop 대한양계협회 2012 월간 양계 Vol.44 No.8

본고는 서울대학교 한재용 교수 연구팀(공동 연구자 박태섭 박사)이 닭에서 바이러스를 사용하지 않은 유전자 전이방법을 이용한 효율적인 형질전환 닭 생산 기술 확립에 성공하여 미국 학술원 회보(Proceedings of the National Academy of Sciences)에 게재함에 따라 그 성과를 기리고 독자들에게 알리기 위해 한재용 교수에 의뢰해 농가들이 알기쉽게 정리한 내용이다. 형질전환 닭은 인간의 질병 연구 및 새로운 치료제 개발을 위한 다양한 실험 모델 생산에 활용되어 양계산업에 다양하게 사용될 것으로 기대되고 있다.

육종 ; Chicken embryo fibroblast 를 이용한 plasmid DNA transfection 에 관한 연구

한재용,서동삼,송기덕,김진원 한국동물자원과학회 ( 구 한국축산학회 ) 1993 한국축산학회 심포지움자료집 Vol.- No.-

형질전환 조류 생산에 관한 연구

한재용 서울대학교 농업개발연구소 1999 농업생명과학연구 Vol.3 No.-

Embryonic germ (EG) cell lines established from primordial germ cells(PGCs) are undifferentiated and pluripotent stem cells. To date, EG cells with proven germ-line transmission have been completely established only in the mouse with embryonic stem (ES) cells. PGCs from 5.5-day-old(stage 28) chicken embryonic gonads were isolated and established a putative chicken EG cell line with EG culture medium supplemented with several growth factors. These cells grew continuously for 10 passages (4 months) on a feeder layer of mitotically active chicken embryonic fibroblasts. After several passages, these cells were characterized by screening with the Periodic acid-Shiff's reaction, anti-SSEA-1 antibody, and a proliferation assay. The chicken EG cells maintained characteristics of gonadal PGCs and undifferentiated stem cells. When cultured in suspension, the chicken EG cells successfully formed an embryoid body and differentiated into a variety of cell types. The chicken EG cells were injected into stage X blastodermal layer and produced chimeric chickens with various differentiated tissues derived from the EG cells. Chicken EG cells will be useful for the production of transgenic chickens and for studies of germ cell differentiation and genomic imprinting. Also, the production of transgenic chicken using these cells may be useful tools for improvement of ecionomic traits in livestocks.

高等學校 日本語敎育의 問題點

韓再龍 경남대학교 교육문제연구소 1992 敎育理論과 實踐 Vol.2 No.-

닭의 다능성 생식세포주를 이용한 생식선 카이메라 생산에 관한 연구

한재용 서울대학교 농업개발연구소 2001 농업생명과학연구 Vol.5 No.-

Researches on manipulating pluripotent stem cells derived from embryos and fetal tissues have a great impact on developing innovative technologies in various fields of life science including agriculture, and biotechnology. Since the first isolation in the mouse embryos, stem cell-like colonies have been continuously established in several species. However, full-term development originated from established pluripotent cells, which is an absolute criterion for proving cell plasticity and differentiation, was only reported in the mouse. Due to technical difficulties, no further progress has been made in the establishment of animal embryonic stem(ES) cell line and the use of embryonic germ (EG) cells is another choice to establish stem cell line especially in the livestock. Avian transgenesis has an unlimited value in biotechnology industry, since its applicability as a bioreactor has proven to be greater than that of mammalian species. In the chicken, EG cells can be extensively utilized instead of ES cells for efficient inducing transgenesis mediated by germline transmission. this study first reported chicken germline chimera production by embryonic germ (EG) cell transplantation into recipient embryos. EG cells were derived from the culture of White Leghorn (WL) gonadal primordial germ cells (gPGCs) at the stage 28. Transplantation was conducted with Korean Ogol chicken embryos at the stage 17. In subsequent testcross analysis, EG cells or gPGCs transplantation yielded total 17 germline chimeras; 2 out of 15 matured progenies in EG cell (13.3%) and 15 of 176 in gPGC cultured for 2 days or fully colonized (8.5%). The efficiency of germline transmission was 1.5 to 14.6% in EG cells, while 0.2 to 27.6% in gPGCs. In conclusion, this is the first report to produce germline chimeras from gPGC-derived EG cells in non-mammalian species. The results of our experiments further demonstrated that chicken EG cells can be established from the culture of gPGCs and that maintained cells can induce germline transmission after transplantation into recipient embryos. Therefore, acquired knowledge on the establishment of EG cell colonies from retrieved gPGCs and confirmation of their pluripotency in this study greatly contribute to accelerating not only transgenesis but also germ cell studies.

자동직경제어 Czochralski 성장장치의 제작

한재용,박성수,이성국,마동준,김용훈,이상학,Han, J.Y.,Park, S.S.,Lee, S.K.,Ma, D.J.,Kim, Y.H.,Lee, S.H. 한국결정성장학회 1994 한국결정성장학회지 Vol.4 No.4

무게 측정에 의한 자동 직경 제어 방식의 Czochralski 단결정 성장 장칭를 제작하였다. 성장로의 몸체와 hot zone을 자체 설계, 제작하였으며, computer에 의하여 전자저울과 유도가열장치를 자동 제어하므로써 가열 공정에서 부터 냉각 공정까지의 모든 과정을 자동화시킬 수 있는 program을 개발하였다. 제작된 성장장치를 사용하여 $LiNbO_3$ 단결정을 성장시켜 성장 장치의 성능을 평가 하였다. We developed a computer controlled Czochralski puller which automated entire crystal growing processes. Crystal weighing technique was employed for an automatic diameter control. The performance of the system was evaluated by the growth of $LiNbO_3$ single crystals.

형질전환 조류 생산에 관한 연구

한재용 서울대학교 농업개발연구소 2000 농업생명과학연구 Vol.4 No.-

Embryonic germ (EG) cell lines established from primordial germ cells (PGCs) are undifferentiated and pluripotent stem cells. To date, EG cells with proven germ-line transmission have been completely established only in the mouse with embryonic stem (ES) cells. PGCs from 5.5-day-old (stage 28) chicken embryonic gonads were isolated and established a putative chicken EG cell line with EG culture medium supplemented with several growth factors. These cells grew continuously for 10 passages (4 months) on a feeder layer of mitotically active chicken embryonic fibroblasts. After several passages, these cells were characterized by screening with the Periodic acid-Shiff's reaction, anti-SSEA-1 antibody, and a proliferation assay. The chicken EG cells successfully formed an embryoid body and differentiated into a variety of cell types. The chicken EG cells were injected into stage X blastodermal layer and produced chimeric chickens with various differentiated tissues derived from the EG cells. Chicken EG cells will be useful for the production of transgenic chickens and for studies of germ cell differentiation and genomic imprinting. We also could produce the germline chimeras using long-term cultured gPGCs without passages. It would be promising strategy to produce transgenic poultry for the improvement of economic traits in livestock.

대형 KTP 단결정 성장 및 광학적 불균일성에 관한 연구

한재용,이성국,마동준,김용훈,박성수,이상학,Han, J.Y.,Lee, S.K.,Ma, D.J.,Kim, Y.H.,Park, S.S.,Lee, S.H. 한국결정성장학회 1994 한국결정성장학회지 Vol.4 No.1

고온용액법(high temperature solution grwth)에 의해 $K_6P_4O_13$의 flux로 부터 KTP 단결정을 성장시켰다. Inclusion이 없는 KTP 단결정을 성장시키기 위해 성방로내 온도기울기, 결정의 회전, 종자결정의 방위, 냉각속도를 조절하였다. 성장된 KTP 단결정은 inclusion이 없었으며, 크기는 $10(a){\times}28(b){\times}33(c)mm^3$ 이었다. 또한 KTP 단결정 boule의 위치에 따른 SHG 출력 특성 측정 및 TEM 분석에 의해 종자결정 주위의 광학적 불균일성의 원인을 조사하였다. Single crystals of Potassium Titanyl Phosphate (KTP) were grown from the flux of $K_6P_4O_13(K_6)$ using a high temperatures solution growth method. To grow the large KTP crystal without inclusion, the temperature gradient in furnace, crystal rotation, orientation of seed crystal, and the cooling rate were controled. The KTP crystals are up to $10(a){\times}28(b){\times}33(c)mm^3$ in size. We investigated the optical inhomogeneity in this KTP crystal by the SHG power measurement and TEM analysis.

형질전환 닭 생산에 관한 연구

한재용 서울대학교 농업생명과학대학 농업개발연구소 1998 농업생명과학연구 Vol.2 No.-

Production of transgenic chicken may be useful tool for studying the expression and function of genes and improvement of economic traits in livestocks. Primordial germ cells (PGCs) have all the genomic information of an individual species and pass this information next generation. Gonadal PGCs were isolated from 6-day-old embryos, transfected with lacZ gene carrying CMV promoter. The addition of DMSO significantly increased the transfection efficiency of electropoartion into gPGCs. Electroporation of gPGCs resulted in an 80% transfection efficiency compared with liposomes (17%). After transfection and reintroduction of gPGCs into recipient embryos, the exogenous gene was detectable in various developmental stage in recipient embryos. These results indicated that electroporation was a suitable means of transfecting avian gPGCs for the goal of producing transgenic poultry.

중심장액맥락망막병증 환자에 대한 스피로놀락톤의 치료 효과 및 안전성

한재용,최은영,이승규 대한안과학회 2020 대한안과학회지 Vol.61 No.11

목적: 중심장액맥락망막병증 환자에서 경구 스피로놀락톤의 치료 효과와 그 안정성에 대해 분석하고자 하였다. 대상과 방법: 중심장액맥락망막병증 진단을 받은 환자들 중 2015년 12월부터 2018년 11월까지 경구 스피로놀락톤을 처방 받은 환자들의 의무기록 및 영상검사를 후향적으로 분석하였다. 환자들의 약제 복용 전과 복용 후 1, 3, 6, 12개월에 중심황반두께, 망막하액 높이, 황반하 맥락막두께 및 최대교정시력을 조사하였고, 환자군을 급성과 만성의 두 군으로 나누어 약제 복용 전후를 비교하였다. 추가로 약제 부작용 및 완전 관해 후 재발된 경우를 조사하였다. 결과: 경구 스피로놀락톤을 복용한 환자 42명의 45안이 포함되었다. 중심황반두께와 망막하액 높이는 모든 경과 관찰 기간에서 치료 전과 비교해 유의하게 호전된 것을 확인하였고, 황반하 맥락막두께의 경우 치료 후 3, 6, 12개월에 유의하게 호전되었다. 하지만 최대교정시력의 경우 모든 경과 관찰 기간에서 치료 전과 유의한 차이가 없었다. 급성 환자군은 전체 환자군과 유사한 경과를 보였고, 만성 환자군의 경우 각 추적 관찰 기간 동안 부분적으로 치료 전후에 차이를 보였다. 완전 관해를 보인 환자 중 8.9%에서 재발을 경험했고, 여성형 유방이 발생한 남자 환자는 2명이었으며, 혈청 크레아티닌 수치의 상승을 보인 환자는 1명이었다. 결론: 이 연구를 통하여 중심장액맥락망막병증 환자에서 스피로놀락톤의 해부학적 구조에 대한 치료 효과를 확인할 수 있었다. 그러나 약제 복용 후 부작용 및 재발의 위험성이 있어 지속적인 경과 관찰이 필요하다. Purpose: To evaluate the therapeutic effects and safety of oral spironolactone (SPRL) in patients with central serous chorioretinopathy (CSC). Methods: Medical records and imaging data from consecutive patients diagnosed with CSC and treated with SPRL were reviewed. Changes in central macular thickness (CMT), subretinal fluids (SRF), subfoveal choroidal thickness (SFCT), and best corrected visual acuity (BCVA) were measured at baseline and follow-up visits. Follow-up visits were performed at 1, 3, 6, and 12 months after treatment. The patients were divided into two groups; acute and chronic, and the therapeutic outcomes were compared. The occurrence of side effects and recurrence after complete resolution of SRF were also assessed. Results: A total of 42 patients with 45 eyes were included. CMT and SRF after using SPRL improved significantly at 1, 3, 6, and 12 months and SFCT improved at 3, 6, and 12 months. However, BCVA showed no significant change at any time. Acute patients showed similar results. However, in chronic patients, CMT showed significant differences at 1 and 3 months, SRF at 1 and 6 months, and SFCT at 6 months. A total of 15.6%, 38.2%, 45.8%, and 50.0% of eyes showed complete resolution of SRF at each follow-up period, respectively, and 8.9% of eyes showed recurrence after complete resolution. Two male patients experienced gynecomastia and one patient showed mild elevation of serum creatinine. Conclusions: Our study supported the therapeutic benefits of SPRL in patients with CSC, especially in anatomical structures. However, careful observation is required because of side effects and recurrence after complete resolution.