Target gene experiment description: |
We also examined the luciferase activity in linearized constructs. If hCNS1 was actually a DPE, it might drive the transcription of the entire sequence in a circular plasmid, which might affect the luciferase gene expression.To prevent hCNS1 from driving the luciferase gene transcription in the sense direction, we linearized Pro--luc--hCNS1 and Pro--luc--rev hCNS1 constructs by cutting the immediate downstream sequence of hCNS1 and rev hCNS1. As a result, hCNS1 still significantly increased TCAM1P promoter activity in HEK293T cells, and the fold--increases were around 3 in both constructs compared to the linearized TCAM1P--Pro--luc construct (Fig. 3B). These data showed that hCNS1 could function as an enhancer for the TCAM1P gene.;We also examined the luciferase activity in linearized constructs. If hCNS1 was actually a DPE, it might drive the transcription of the entire sequence in a circular plasmid, which might affect the luciferase gene expression.To prevent hCNS1 from driving the luciferase gene transcription in the sense direction, we linearized Pro--luc--hCNS1 and Pro--luc--rev hCNS1 constructs by cutting the immediate downstream sequence of hCNS1 and rev hCNS1. As a result, hCNS1 still significantly increased TCAM1P promoter activity in HEK293T cells, and the fold--increases were around 3 in both constructs compared to the linearized TCAM1P--Pro--luc construct (Fig. 3B). These data showed that hCNS1 could function as an enhancer for the TCAM1P gene. |