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Poster Presentation : Gene Expression / Function

Genome Editing in Chicken Primordial Germ Cells by CRIPSR-Cas9 System

첫 페이지 보기
  • 발행기관
    한국동물생명공학회(구 한국동물번식학회) 바로가기
  • 간행물
    발생공학 국제심포지엄 및 학술대회 바로가기
  • 통권
    The 17th International Symposium on Developmental Biotechnology (2017.10)바로가기
  • 페이지
    pp.43-43
  • 저자
    Jeong Hyo Lee, Jeong-Woong Park, Tae Sub Park
  • 언어
    영어(ENG)
  • URL
    https://www.earticle.net/Article/A347324

원문정보

초록

영어
To date, CRISPR/CRISPR-associated protein 9 (Cas9)-mediated genome editing technology is expected to become the most efficient tool for economic trait improvement in livestock. However, one of the critical issues for genetic modification by CRISPR/Cas9 is non-specific DNA double strand breaks which is called as off-target effect. In this study, we examined the mutated Cas9, nickase to modulate the specific target gene in chicken primordial germ cells (PGCs) as well as DF1 cells. Chicken myostatin which is famous for inhibitor of muscle cell growth and differentiation during myogenesis was targeted to be mutated with the Cas9-D10A nickase. After co-transfection of the Cas9- D10A nickase expression vector with eGFP gene and two targeted guide RNAs (g- RNAs), the GFP-positive cells were sorted out by fluorescence-activated cell sorting (FACS). In DF1 cells which is chicken embryonic fibroblast cell line, the mutant induction efficiency was 100% in the targeted myostatin site by the genotyping analysis of the knockout DF1 cells. In DF1 cells, number of the deleted nucleotides ranged from 2 to 39 nucleotide deletion. In addition, 6 expected off-target sites were predicted and analyzed but any non-specific mutation in the off-target sites was not observed. We also verified myostatin gene knockout with Cas9-D10A nickase in chicken primordial germ cells (PGCs). In conclusion, the knockout technical platform with the Cas9- D10A nickase can be efficiently applied to functional genomics study in poultry and finally adapted to generate the knockout poultry.

저자

  • Jeong Hyo Lee [ Graduate School of International Agricultural Technology ]
  • Jeong-Woong Park [ Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea ]
  • Tae Sub Park [ Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea ]

참고문헌

자료제공 : 네이버학술정보

간행물 정보

발행기관

  • 발행기관명
    한국동물생명공학회(구 한국동물번식학회) [The Korean Society of Animal Reproduction and Biotechnology]
  • 설립연도
    1976
  • 분야
    농수해양>축산학
  • 소개
    동물번식생리학, 동물생명공학, 수의학, 인공수정 및 수정란이식을 이용한 동물개량에 관한 이론과 기술의 발전을 통해 학계, 연구계, 산업계 및 양축가 상호간의 협력을 도모함으로써 동물과학발전 및 사회일반의 이익에 기여 한다는 목적을 위해 노력해 나가겠습니다.

간행물

  • 간행물명
    발생공학 국제심포지엄 및 학술대회 [International Symposium on Developmental Biotechnology]
  • 간기
    연간
  • 수록기간
    2004~2018
  • 십진분류
    KDC 527 DDC 636

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