지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2019.1
- 수록면
- 418 - 425 (8page)
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초록· 키워드
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Multicistronic elements, such as the internal ribosome entry site (IRES) and 2A-like cleavage sequence, serve crucial roles in the eukaryotic ectopic expression of exogenous genes. For utilization of multicistronic elements, the cleavage efficiency and order of elements in multicistronic vectors have been investigated; however, the dynamics of multicistronic element-mediated expression remains unclear. Here, we investigated the dynamics of encephalomyocarditis virus (EMCV) IRES- and porcine teschovirus-1 2A (p2A)-mediated expression. By utilizing real-time fluorescent imaging at a minute-level resolution, we monitored the expression of fluorescent reporters bridged by either EMCV IRES or p2A in two independent cultured cell lines, HEK293 and Neuro2a. We observed significant correlations for the two fluorescent reporters in both multicistronic elements, with a higher correlation coefficient for p2A in HEK293 but similar coefficients for IRES-mediated expression and p2A-mediated expression in Neuro2a. We further analyzed the causal relationship of multicistronic elements by convergent cross mapping (CCM). CCM revealed that in all four conditions examined, the expression of the preceding gene causally affected the dynamics of the subsequent gene. As with the cross correlation, the predictive skill of p2A was higher than that of IRES in HEK293, while the predictive skills of the two multicistronic elements were indistinguishable in Neuro2a. To summarize, we report a significant temporal correlation in both EMCV IRES- and p2A-mediated expression based on the simple bicistronic vector and real-time fluorescent monitoring. The current system also provides a valuable platform to examine the dynamic aspects of expression mediated by diverse multicistronic elements under various physiological conditions.
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참고문헌 (34)
참고문헌 신청
Arnaud, O. / 2015 / Temperature-induced variation in gene expression burst size in metazoan cells / BMC Mol. Biol 16:20
Balvay, L. / 2009 / Structural and functional diversity of viral IRESes / Biochim. Biophys. Acta 1789:542~557
Bouabe, H. / 2008 / Improvement of reporter activity by IRES-mediated polycistronic reporter system / Nucleic Acids Res 36:e28
Daigle, T. L. / 2018 / A suite of transgenic driver and reporter mouse lines with enhanced brain-cell-type targeting and functionality / Cell 174:465.e22~480.e22
Evdokimov, A. G. / 2006 / Structural basis for the fast maturation of Arthropoda green fluorescent protein / EMBO Rep 7:1006~1012
Balvay, L. / 2009 / Structural and functional diversity of viral IRESes / Biochim. Biophys. Acta 1789:542~557
Bouabe, H. / 2008 / Improvement of reporter activity by IRES-mediated polycistronic reporter system / Nucleic Acids Res 36:e28
Daigle, T. L. / 2018 / A suite of transgenic driver and reporter mouse lines with enhanced brain-cell-type targeting and functionality / Cell 174:465.e22~480.e22
Evdokimov, A. G. / 2006 / Structural basis for the fast maturation of Arthropoda green fluorescent protein / EMBO Rep 7:1006~1012
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