지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제22권 제2호
- 발행연도
- 2012.1
- 수록면
- 190 - 198 (9page)
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초록· 키워드
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RNA interference (RNAi) is rapidly becoming a valuable tool in biological studies, as it allows the selective and transient knockdown of protein expression. The shortinterfering RNAs (siRNAs) transiently silence gene expression. By contrast, the expressed short-hairpin RNAs induce long-term, stable knockdown of their target gene. Trichoplusia ni (T. ni) cells are widely used for mammalian cell-derived glycoprotein expression using the baculovirus system. However, a suitable shRNA expression system has not been developed yet. We investigated the potency of shRNA-mediated gene expression inhibition using human and Drosophila U6promoters in T. ni cells. Luciferase, EGFP, and β-Nacetylglucosaminidase (GlcNAcase) were employed as targets to investigate knockdown of specific genes in T. ni cells.
Introduction of the shRNA expression vector under the control of human U6 or Drosophila U6 promoter into T. ni cells exhibited the reduced level of luciferase, EGFP, and β-Nacetylglucosaminidase compared with that of untransfected cells. The shRNA was expressed and processed to siRNA in our vector-transfected T. ni cells. GlcNAcase mRNA levels were down-regulated in T. ni cells transfected with shRNA vectors-targeted GlcNAcase as compared with the control vector-treated cells. It implied that our shRNA expression vectors using human and Drosophila U6promoters were applied in T. ni cells for the specific gene knockdown.
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참고문헌 (24)
참고문헌 신청
Altman, F. / 1999 / Insect cells as host for the expression of recombinant glycoproteins / Glycoconj. J 16 : 109 ~ 123
Altmann, F. / 1996 / Insect cells contain an unusual membrane-bound β- N-acetylglucosaminidase probably involved in the processing of protein N-glycans / J. Biol. Chem 270 :
Amarzguioui, M. / 2004 / An algorithm for selection of functional siRNA sequences / Biochem. Biophys. Res. Commun 316 : 1050 ~ 1058
Bernstein, E. / 2001 / Role for a bidentate ribonuclease in the initiation step ofRNA interference / Nature 409 : 363 ~ 366
Brummelkamp, T. R. / 2002 / A system for stable expression of short interfering RNAs mammalian cells / Science 296 : 550 ~ 553
Altmann, F. / 1996 / Insect cells contain an unusual membrane-bound β- N-acetylglucosaminidase probably involved in the processing of protein N-glycans / J. Biol. Chem 270 :
Amarzguioui, M. / 2004 / An algorithm for selection of functional siRNA sequences / Biochem. Biophys. Res. Commun 316 : 1050 ~ 1058
Bernstein, E. / 2001 / Role for a bidentate ribonuclease in the initiation step ofRNA interference / Nature 409 : 363 ~ 366
Brummelkamp, T. R. / 2002 / A system for stable expression of short interfering RNAs mammalian cells / Science 296 : 550 ~ 553
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