Cited 4 time in
Cited 4 time in
Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence
- Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence
- Lee, Soo Min; Kim, Jeongah; Hee-Dae Kim; Chung, Sooyoung; Kim, Kyungjin; Choe, Han Kyoung
- DGIST Authors
- Choe, Han Kyoung
- Issue Date
- Molecules and Cells, 42(5), 418-425
- Article Type
- Author Keywords
- 2A; expression dynamics; internal ribosome entry site; multicistronic elements; real-time fluorescent imaging
- CIRCADIAN GENE-EXPRESSION; MESSENGER-RNA; FLUORESCENT PROTEINS; TRANSLATION; BIOLOGY; LIVER
- 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.
- Related Researcher
Choe, Han Kyoung
Laboratory of Animal Behavior and Circadian rhythm
Modulation of neural circuit; Circadian regulation of behavior and perception; Neurotechnology
There are no files associated with this item.
- Department of Brain and Cognitive SciencesLaboratory of Animal Behavior and Circadian rhythm1. Journal Articles
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.