Abstract

Background

Close Homolog of L1 (CHL1) is a member of the L1 family of cell adhesion molecules. CHL1 gene is located on human chromosome 3 and has been linked to several pathologies, including 3p deletion syndrome, schizophrenia, and tumor growth and metastasis.

Objective

The goal of the present study was to determine which region of the CHL1 promoter is most competent in driving CHL1 gene expression. Methods: Five candidate DNA fragments in the promoter regions were selected by screening across six species for evolutionary conserved sequences. The activity of these five promoter regions was quantitatively evaluated using a GFP reporter gene in transfection experiments, performed in C6 glioma cells.

Results

Of the five promoter regions tested, three drove reporter GFP expression, with the conserved region 6 (CR6, Gene ID AC066595.5, 25851-26850) being the most active for transcription.

Conclusion

The identification of the CR6 activity provides a better understanding of the regulatory mechanisms underlying CHL1 expression. It may help future discovery of therapeutic strategies that involve influencing critical promoter regions to drive transcriptional regulation of the mammalian CHL1 gene.

• HIGHLIGHTS

• Conserved regions of CHL1 promoter sequences were identified by in-silico analysis.

• Five conserved regions were tested for gene regulatory activity using a reporter assay.

• Conserved regions CR5, CR6 and CR7 show gene regulatory function in a reporter assay.

• Co-transfection of CR5 and CR6 yielded the highest reporter activity.

• The core region of CR6 (CR6core) was identified as a cis-acting element.

• In-tandem promoter CR5core-CR6core was the best in a reporter assay.

中文翻译：

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人粘附分子L1密切同源物启动子区功能序列分析

摘要

背景

L1 (CHL1) 的紧密同源物是细胞粘附分子 L1 家族的成员。CHL1 基因位于人类第 3 号染色体上，与多种疾病有关，包括 3p 缺失综合征、精神分裂症以及肿瘤生长和转移。

客观的

本研究的目的是确定 CHL1 启动子的哪个区域最有能力驱动 CHL1 基因表达。方法：通过筛选六个物种的进化保守序列，选择了启动子区域中的五个候选 DNA 片段。在 C6 神经胶质瘤细胞中进行的转染实验中，使用 GFP 报告基因对这五个启动子区域的活性进行了定量评估。

结果

在测试的五个启动子区域中，三个驱动报告基因 GFP 表达，其中保守区域 6（CR6，基因 ID AC066595.5, 25851-26850）对转录最活跃。

结论

CR6 活性的鉴定提供了对 CHL1 表达的调控机制的更好理解。它可能有助于未来发现涉及影响关键启动子区域以驱动哺乳动物CHL1基因转录调控的治疗策略。

• 强调

• 通过计算机分析鉴定 CHL1 启动子序列的保守区域。

• 使用报告分析测试了五个保守区域的基因调控活性。

• 保守区 CR5、CR6 和 CR7 在报告基因测定中显示出基因调控功能。

• CR5 和 CR6 的共转染产生了最高的报告活性。

• CR6 (CR6core) 的核心区域被确定为顺式作用元件。

• 串联启动子 CR5core-CR6core 在报告基因分析中是最好的。