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NTIS 바로가기Journal of plant biotechnology = 식물생명공학회지, v.45 no.3, 2018년, pp.155 - 170
In this paper, mechanisms of gene editing technologies including ZFN, TALENS and CRISPR were briefly discussed with mutual advantages and disadvantages. Classification criteria of gene edited, site-directed mutagenesis (SDN) crops for regulatory purpose were also discussed. The number of studies usi...
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핵심어 | 질문 | 논문에서 추출한 답변 |
---|---|---|
CRISPR란? | 3세대 기술인 CRISPR기술은 절단하고자 하는 DNA영역을 single-guide RNA (sgRNA)가 인식한다는 점이 앞서 설명한 두 가지 기술과 비교하여 가장 크게 다르다. CRISPR라는 용어는 Clustered Regulatory Inter-spaced Short Palindromic Repeat의 약어로서 Jansen 등(2002)이 발표한 논문에 처음 사용되었는데 대장균의 iap유전자의 하위영역에 존재하는 비 반복서열 다음에 인접해 있는 tandem repeat를 지칭한다(Ishino et al.1987). | |
CRISPR-Cas9 또는 Cpf1 시스템을 동물 또는 식물 등의 생물체에 차용하기 위해 필요한 것은? | CRISPR-Cas9 또는 Cpf1시스템을 동물 또는 식물 등의 생물체에 차용하기 위하여 다음과 같은 실험 단계들 즉 목표로하는 유전자 내 Protospacer adjacent motif (PAM)염기서열 확인, 단일가닥 RNA (sgRNA)합성, 적합한 운반체 내 클로닝, 숙주세포로의 도입, 스크리닝, 검정 등이 필요하다. CRISPR기술의 가장 큰 장점은 PAM영역의 다양한 돌연변이 유발, target RNA의 합성, 운반체 내 클로닝 등의 바이오 부품 제작 등의 기술이 지속적으로 개발되고 있어 제작이 용이하고 효율성과 정확도가 높다는 점이다. | |
EFSA는 안전성평가를 용이하게 하기 위하여 ZFN 기술을 어떻게 분류하고 정의하였는가? | European Food Safety Authority (EFSA, 2012)는 안전성평가를 용이하게 하기 위하여 ZFN 기술을 ZFN-1, ZFN-2, ZFN-3기술로 분류하고 각각을 다음과 같이 정의하였다. ZFN-1은 NHEJ기작에 의하여 단일염기의 변화, 짧은 염기의 삽입 또는 결실이 일어난 것으로 이중나선의 절단 후 교정과정에서 주형가닥(repair template)을 사용하지 않아야 한다고 정의하였다. 또한 삽입이 일어난 경우에 삽입된 염기는 반드시 자신의 게놈으로 부터 유래된 것이어야 하고 외부에서 삽입되지 않아야 한다. ZFN-2는 상동성재조합기작(homologous recombination, HR)에 의하여 특정단일 또는 작은 수의 염기가 변화, 삽입, 결실 등이 일어나는 점 돌연변이의 경우로 한정하였다. ZFN-2는 점 돌연변이를 도입하기 위하여 target DNA와 상동성이면서 도입하기를 원하는 점 돌연변이를 포함하는 DNA주형가닥을 사용한다. ZFN-3는 상동성재조합(HR)기작을 이용하여 외래 DNA단편 또는 유전자 카셋트를 도입한 것으로 정의하였다. 이때 도입된 DNA단편의 길이는 수 천 염기쌍(kilo base pairs, kbps)까지 포함한다고 하였다. 그리고 실제로는 HR뿐만 아니라 NHEJ 기작에 의하여도 특정 외래 DNA단편의 도입도 일어날 수 있으며 이 경우를 특별히 ZFN3-NHEJ로 구분하였다. EFSA는 이러한 등급 구분 체계는 TALENS와 CRISPR 그리고 앞으로 새로이 개발되는 모든 유전자편집기술에도 동일하게 적용하여 이들 site-directednuclease (SDN)기술을 SDN-1, SDN-2, SDN-3로 구분하고 특히 SDN-3에 해당하는 것들은 외래 DNA단편을 사전에 알고 있는 위치(predefined region)에 삽입한다는 것 외에는 기존의 유전자변형생물체와 동일한 것으로 간주하고 안전성평가과정을 필하여야 한다고 주장하였다. |
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