Cited 4 time in webofscience Cited 4 time in scopus

Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence

Title
Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence
Authors
Lee, Soo MinKim, JeongahHee-Dae KimChung, SooyoungKim, KyungjinChoe, Han Kyoung
DGIST Authors
Choe, Han Kyoung
Issue Date
2019-05
Citation
Molecules and Cells, 42(5), 418-425
Type
Article
Article Type
Article
Author Keywords
2Aexpression dynamicsinternal ribosome entry sitemulticistronic elementsreal-time fluorescent imaging
Keywords
CIRCADIAN GENE-EXPRESSIONMESSENGER-RNAFLUORESCENT PROTEINSTRANSLATIONBIOLOGYLIVER
ISSN
1016-8478
Abstract
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.
URI
http://hdl.handle.net/20.500.11750/9904
DOI
10.14348/molcells.2019.2427
Publisher
한국분자세포생물학회
Related Researcher
  • Author Choe, Han Kyoung Laboratory of Animal Behavior and Circadian rhythm
  • Research Interests Modulation of neural circuit; Circadian regulation of behavior and perception; Neurotechnology
Files:
There are no files associated with this item.
Collection:
Department of Brain and Cognitive SciencesLaboratory of Animal Behavior and Circadian rhythm1. Journal Articles


qrcode mendeley

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE