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Upregulation of endogenous HMOX1 expression by a computer-designed artificial transcription factor.

Guo H, Tian Y, Lu H, Wei Y, Ying D - J. Biomed. Biotechnol. (2010)

Bottom Line: In the ATF, the p65 functional domain was used as the effector domain (ED), and a nuclear localization sequence (NLS) was also included.We next analyzed the affinity of the ATF to the HMOX1 enhancer and the effect of the ATF on endogenous HMOX1 expression.The results suggest that the ATF could effectively upregulate endogenous HMOX1 expression in ECV304 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Third Military Medical University, Sha-Ping-Ba District, Chongqing, China.

ABSTRACT
Heme oxygenase-1 (HO-1) is well known as a cytoprotective factor. Research has revealed that it is a promising therapeutic target for cardiovascular diseases. In the current study, an HMOX1 (HO-1 gene) enhancer-specific artificial zinc-finger protein (AZP) was designed using bioinformatical methods. Then, an artificial transcription factor (ATF) was constructed based on the AZP. In the ATF, the p65 functional domain was used as the effector domain (ED), and a nuclear localization sequence (NLS) was also included. We next analyzed the affinity of the ATF to the HMOX1 enhancer and the effect of the ATF on endogenous HMOX1 expression. The results suggest that the ATF could effectively upregulate endogenous HMOX1 expression in ECV304 cells. With further research, the ATF could be developed as a potential drug for cardiovascular diseases.

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Related in: MedlinePlus

The DNA sequence selected for prescreening. A 54 bp DNA sequence between the two stress response elements (StREs) of human HMOX1 enhancer (underlined) was selected for prescreening.
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fig1: The DNA sequence selected for prescreening. A 54 bp DNA sequence between the two stress response elements (StREs) of human HMOX1 enhancer (underlined) was selected for prescreening.

Mentions: The sequence between the two stress response elements (StREs) of human HMOX1 enhancer was submitted to ZF tools 3.0 to search AZP target sites (Figure 1). Parameter settings are (1) “Triplets to search” included all available triplets (GNG, GN(ACT), ANN, CNN, and TNN). (2) “ZF set” was “total” and “Contiguous targets” radio button was selected. (3) “Minimum target size” was “18”. After that, several candidate target sites were found.


Upregulation of endogenous HMOX1 expression by a computer-designed artificial transcription factor.

Guo H, Tian Y, Lu H, Wei Y, Ying D - J. Biomed. Biotechnol. (2010)

The DNA sequence selected for prescreening. A 54 bp DNA sequence between the two stress response elements (StREs) of human HMOX1 enhancer (underlined) was selected for prescreening.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2913762&req=5

fig1: The DNA sequence selected for prescreening. A 54 bp DNA sequence between the two stress response elements (StREs) of human HMOX1 enhancer (underlined) was selected for prescreening.
Mentions: The sequence between the two stress response elements (StREs) of human HMOX1 enhancer was submitted to ZF tools 3.0 to search AZP target sites (Figure 1). Parameter settings are (1) “Triplets to search” included all available triplets (GNG, GN(ACT), ANN, CNN, and TNN). (2) “ZF set” was “total” and “Contiguous targets” radio button was selected. (3) “Minimum target size” was “18”. After that, several candidate target sites were found.

Bottom Line: In the ATF, the p65 functional domain was used as the effector domain (ED), and a nuclear localization sequence (NLS) was also included.We next analyzed the affinity of the ATF to the HMOX1 enhancer and the effect of the ATF on endogenous HMOX1 expression.The results suggest that the ATF could effectively upregulate endogenous HMOX1 expression in ECV304 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Third Military Medical University, Sha-Ping-Ba District, Chongqing, China.

ABSTRACT
Heme oxygenase-1 (HO-1) is well known as a cytoprotective factor. Research has revealed that it is a promising therapeutic target for cardiovascular diseases. In the current study, an HMOX1 (HO-1 gene) enhancer-specific artificial zinc-finger protein (AZP) was designed using bioinformatical methods. Then, an artificial transcription factor (ATF) was constructed based on the AZP. In the ATF, the p65 functional domain was used as the effector domain (ED), and a nuclear localization sequence (NLS) was also included. We next analyzed the affinity of the ATF to the HMOX1 enhancer and the effect of the ATF on endogenous HMOX1 expression. The results suggest that the ATF could effectively upregulate endogenous HMOX1 expression in ECV304 cells. With further research, the ATF could be developed as a potential drug for cardiovascular diseases.

Show MeSH
Related in: MedlinePlus