基于您提供的文件列表和已发表的论文背景,以下是 LT (Large Tumor Antigen) 相关未发表数据的完整表格整理。这些数据分为 ChIP-seq(LT蛋白结合位点)和 RNA-seq(LT及突变体转录组影响)。
1. LT 蛋白特异性 ChIP-Seq 数据集
状态: 未发表 (Unpublished)
目的: 绘制 LT 蛋白在全基因组上的直接结合位点。
抗体: Cm2b4 (针对 LT/LTtr 蛋白) 或 IgG/Input 对照。
注意: 部分样本为 LT+sT 共表达,部分为 LT 单独表达。
| 样本名称 (Sample ID) | 细胞类型 | 供体/ID | 处理条件 (Condition) | 对照类型 (Control) | 文件名示例 (File Name) | 备注 |
|---|---|---|---|---|---|---|
| HEK293_r1 | HEK293 | N/A | LT + sT | Input | HEK293_LT+sT_r1.fastq.gzHEK293_LT+sT_r1_Input.fastq.gz |
重复1 |
| HEK293_r2 | HEK293 | N/A | LT + sT | Input | HEK293_LT+sT_r2.fastq.gz |
重复2 (Input可能在其他目录或共用) |
| HEK293_r3 | HEK293 | N/A | LT + sT | Input | HEK293_LT+sT_r3.fastq.gzHEK293_LT+sT_r3_Input.fastq.gz |
重复3 |
| HEK293_Mock_r1-3 | HEK293 | N/A | Mock (Vector) | Input | HEK293_mock_r1...r3.fastq.gz..._Input.fastq.gz |
阴性对照 (之前分析中 r2/r3 质量较差可能被排除) |
| hTERT_LT_r1 | hTERT (BJ5ta) | N/A | LT only | Input? | hTERT_LT_r1.fastq.gz |
仅 LT 表达 (注意:之前分析中此样本可能因质量被排除,需检查对应 Input) |
| hTERT_LT_r2 | hTERT (BJ5ta) | N/A | LT only | Input | hTERT_LT_r2.fastq.gzhTERT_LT_r2_Input.fastq.gz |
仅 LT 表达,高质量重复 |
| hTERT_LT+sT_r1 | hTERT (BJ5ta) | N/A | LT + sT | Input | hTERT_LT+sT_r1.fastq.gzhTERT_LT+sT_r1_Input.fastq.gz |
LT+sT 共表达 |
| hTERT_LT+sT_r2 | hTERT (BJ5ta) | N/A | LT + sT | Input | hTERT_LT+sT_r2.fastq.gzhTERT_LT+sT_r2_Input.fastq.gz |
LT+sT 共表达 |
| hTERT_Mock_r1-2 | hTERT (BJ5ta) | N/A | Mock (Vector) | Input | hTERT_mock_r1...r2.fastq.gz..._Input.fastq.gz |
阴性对照 |
| NHDF_Donor1 | NHDF (Primary) | Donor 1 | LT (Cm2b4) | Input | NHDF_LT_Donor1.fastq.gzNHDF_LT_Donor1_Input.fastq.gz |
原代细胞,生理相关性高 |
| NHDF_Donor2 | NHDF (Primary) | Donor 2 | LT (Cm2b4) | Input | NHDF_LT_Donor2.fastq.gzNHDF_LT_Donor2_Input.fastq.gz |
原代细胞,生理相关性高 |
| p783_DonorI | NHDF (Primary) | Donor I (p783) | LT?/Cm2b4 | Input | p783_ChIP_DonorI.fastq.gzp783_input_DonorI.fastq.gz |
2023年新批次数据 |
| p783_DonorII | NHDF (Primary) | Donor II (p783) | LT?/Cm2b4 | Input | p783_ChIP_DonorII.fastq.gzp783_input_DonorII.fastq.gz |
2023年新批次数据 |
| PFSK-1A_r1 | PFSK-1A | N/A | LT + sT | IgG | PFSK-1A_LT+sT_r1.fastq.gzPFSK-1A_LT+sT_r1_IgG.fastq.gz |
注意: 使用 IgG 作为对照,而非 Input |
| PFSK-1A_r2 | PFSK-1A | N/A | LT + sT | IgG | PFSK-1A_LT+sT_r2.fastq.gzPFSK-1A_LT+sT_r2_IgG.fastq.gz |
注意: 使用 IgG 作为对照 |
| PFSK-1B_r1 | PFSK-1B | N/A | LT + sT | Input | PFSK-1B_LT+sT_r1.fastq.gzPFSK-1B_LT+sT_r1_Input.fastq.gz |
使用 Input 作为对照 |
| PFSK-1B_r2 | PFSK-1B | N/A | LT + sT | Input | PFSK-1B_LT+sT_r2.fastq.gzPFSK-1B_LT+sT_r2_Input.fastq.gz |
使用 Input 作为对照 |
| PFSK-1A_H3K4 | PFSK-1A | N/A | H3K4me3 ChIP | IgG | PFSK-1A_H3K4_...fastq.gz |
非LT蛋白ChIP,为组蛋白修饰数据,可用于辅助注释 |
2. LT-K331A 突变体 RNA-Seq 数据集
状态: 未发表 (Unpublished) 目的: 比较野生型 LT、截短体 LTtr 和解旋酶突变体 K331A 对宿主转录组的影响。 细胞模型: NHDF (原代人真皮成纤维细胞),两个独立供体 (Donor I & II)。 时间点: 转导后第 8 天 (Day 8)。
| 样本名称 (Sample ID) | 细胞类型 | 供体 (Donor) | 处理条件 (Condition) | 文件名 (File Name) | 备注 |
|---|---|---|---|---|---|
| Control_DI | NHDF | Donor I | Vector Control | control_d8_DonorI.fastq.gz |
空白对照 |
| Control_DII | NHDF | Donor II | Vector Control | control_d8_DonorII.fastq.gz |
空白对照 |
| Control_DII_Re | NHDF | Donor II | Vector Control | control-d8-DII_re.fastq.gz |
Donor II 的重复或重测数据 |
| LT_WT_DI | NHDF | Donor I | LT (Wild Type) | LT_d8_DonorI.fastq.gz |
野生型 LT |
| LT_WT_DII | NHDF | Donor II | LT (Wild Type) | LT_d8_DonorII.fastq.gz |
野生型 LT |
| LTtr_DI | NHDF | Donor I | LTtr (Truncated) | LTtr_d8_DonorI.fastq.gz |
肿瘤相关截短体 |
| LTtr_DII | NHDF | Donor II | LTtr (Truncated) | LTtr_d8_DonorII.fastq.gz |
肿瘤相关截短体 |
| LT_K331A_DI | NHDF | Donor I | LT-K331A (Mutant) | LT_K331A_d8_DonorI.fastq.gz |
解旋酶结构域突变体 |
| LT_K331A_DII | NHDF | Donor II | LT-K331A (Mutant) | LT_K331A_d8_DonorII.fastq.gz |
解旋酶结构域突变体 |
| LT_K331A_DII_Re | NHDF | Donor II | LT-K331A (Mutant) | LT-K331A-d8-DII_re.fastq.gz |
Donor II 的重复或重测数据 |
数据分析建议
-
ChIP-Seq 分析策略:
- 分组: 将样本分为
LT_only(hTERT_LT_r2, NHDF_Donors),LT+sT(HEK293, hTERT_LT+sT, PFSK), 和Mock。 - 对照处理: 注意 PFSK-1A 使用 IgG 对照,而其他样本主要使用 Input。在调用峰值 (Peak Calling, 如 MACS2) 时,需针对不同对照类型调整参数或分别处理。
- 整合: 将 LT 结合位点与 RNA-Seq 中的差异基因 (DEGs) 进行重叠分析,特别是关注 LT-K331A 突变是否导致某些关键基因启动子区域的 LT 结合丢失或减弱。
- 分组: 将样本分为
-
RNA-Seq 分析策略:
- 差异表达: 使用 DESeq2 进行以下对比:
LT_WT vs ControlLTtr vs ControlLT_K331A vs ControlLT_K331A vs LT_WT(关键对比:确定 K331A 突变特异性影响的基因)
- 功能富集: 重点关注干扰素信号通路 (ISGs)、细胞周期调控和 DNA 损伤反应基因。根据已发表论文,LT 会诱导 ISGs,而 K331A 突变可能会改变这种诱导能力或影响其他下游通路。
- 差异表达: 使用 DESeq2 进行以下对比:
-
多组学整合:
- 利用 NHDF Donor 1 & 2 的数据进行跨组学整合,因为这部分既有 ChIP-seq (LT 结合) 又有 RNA-seq (LT/K331A 表达影响),且来自相同的原代细胞系统,生物学一致性最高。
Based on the file list you provided and the background of published papers, here is the complete table of unpublished data related to LT (Large Tumor Antigen). These data are divided into ChIP-seq (LT protein binding sites) and RNA-seq (transcriptomic impact of LT and its mutants).
1. LT Protein-Specific ChIP-Seq Datasets
Status: Unpublished
Objective: To map the direct genome-wide binding sites of the LT protein.
Antibody: Cm2b4 (targeting LT/LTtr proteins) or IgG/Input controls.
Note: Some samples involve co-expression of LT+sT, while others involve LT expression alone.
| Sample ID | Cell Type | Donor/ID | Condition | Control Type | Example File Name | Remarks |
|---|---|---|---|---|---|---|
| HEK293_r1 | HEK293 | N/A | LT + sT | Input | HEK293_LT+sT_r1.fastq.gzHEK293_LT+sT_r1_Input.fastq.gz |
Replicate 1 |
| HEK293_r2 | HEK293 | N/A | LT + sT | Input | HEK293_LT+sT_r2.fastq.gz |
Replicate 2 (Input may be in another directory or shared) |
| HEK293_r3 | HEK293 | N/A | LT + sT | Input | HEK293_LT+sT_r3.fastq.gzHEK293_LT+sT_r3_Input.fastq.gz |
Replicate 3 |
| HEK293_Mock_r1-3 | HEK293 | N/A | Mock (Vector) | Input | HEK293_mock_r1...r3.fastq.gz..._Input.fastq.gz |
Negative control (r2/r3 might have been excluded due to poor quality in previous analyses) |
| hTERT_LT_r1 | hTERT (BJ5ta) | N/A | LT only | Input? | hTERT_LT_r1.fastq.gz |
LT expression only (Note: This sample might have been excluded due to quality in previous analyses; check for corresponding Input) |
| hTERT_LT_r2 | hTERT (BJ5ta) | N/A | LT only | Input | hTERT_LT_r2.fastq.gzhTERT_LT_r2_Input.fastq.gz |
LT expression only, high-quality replicate |
| hTERT_LT+sT_r1 | hTERT (BJ5ta) | N/A | LT + sT | Input | hTERT_LT+sT_r1.fastq.gzhTERT_LT+sT_r1_Input.fastq.gz |
Co-expression of LT+sT |
| hTERT_LT+sT_r2 | hTERT (BJ5ta) | N/A | LT + sT | Input | hTERT_LT+sT_r2.fastq.gzhTERT_LT+sT_r2_Input.fastq.gz |
Co-expression of LT+sT |
| hTERT_Mock_r1-2 | hTERT (BJ5ta) | N/A | Mock (Vector) | Input | hTERT_mock_r1...r2.fastq.gz..._Input.fastq.gz |
Negative control |
| NHDF_Donor1 | NHDF (Primary) | Donor 1 | LT (Cm2b4) | Input | NHDF_LT_Donor1.fastq.gzNHDF_LT_Donor1_Input.fastq.gz |
Primary cells, high physiological relevance |
| NHDF_Donor2 | NHDF (Primary) | Donor 2 | LT (Cm2b4) | Input | NHDF_LT_Donor2.fastq.gzNHDF_LT_Donor2_Input.fastq.gz |
Primary cells, high physiological relevance |
| p783_DonorI | NHDF (Primary) | Donor I (p783) | LT?/Cm2b4 | Input | p783_ChIP_DonorI.fastq.gzp783_input_DonorI.fastq.gz |
New batch data from 2023 |
| p783_DonorII | NHDF (Primary) | Donor II (p783) | LT?/Cm2b4 | Input | p783_ChIP_DonorII.fastq.gzp783_input_DonorII.fastq.gz |
New batch data from 2023 |
| PFSK-1A_r1 | PFSK-1A | N/A | LT + sT | IgG | PFSK-1A_LT+sT_r1.fastq.gzPFSK-1A_LT+sT_r1_IgG.fastq.gz |
Note: Uses IgG as control, not Input |
| PFSK-1A_r2 | PFSK-1A | N/A | LT + sT | IgG | PFSK-1A_LT+sT_r2.fastq.gzPFSK-1A_LT+sT_r2_IgG.fastq.gz |
Note: Uses IgG as control |
| PFSK-1B_r1 | PFSK-1B | N/A | LT + sT | Input | PFSK-1B_LT+sT_r1.fastq.gzPFSK-1B_LT+sT_r1_Input.fastq.gz |
Uses Input as control |
| PFSK-1B_r2 | PFSK-1B | N/A | LT + sT | Input | PFSK-1B_LT+sT_r2.fastq.gzPFSK-1B_LT+sT_r2_Input.fastq.gz |
Uses Input as control |
| PFSK-1A_H3K4 | PFSK-1A | N/A | H3K4me3 ChIP | IgG | PFSK-1A_H3K4_...fastq.gz |
Not LT protein ChIP; histone modification data, useful for auxiliary annotation |
2. LT-K331A Mutant RNA-Seq Datasets
Status: Unpublished Objective: To compare the impact of Wild-Type LT, Truncated LT (LTtr), and Helicase Mutant K331A on the host transcriptome. Cell Model: NHDF (Primary Human Dermal Fibroblasts), two independent donors (Donor I & II). Time Point: Day 8 post-transduction.
| Sample ID | Cell Type | Donor | Condition | File Name | Remarks |
|---|---|---|---|---|---|
| Control_DI | NHDF | Donor I | Vector Control | control_d8_DonorI.fastq.gz |
Blank control |
| Control_DII | NHDF | Donor II | Vector Control | control_d8_DonorII.fastq.gz |
Blank control |
| Control_DII_Re | NHDF | Donor II | Vector Control | control-d8-DII_re.fastq.gz |
Replicate or re-test data for Donor II |
| LT_WT_DI | NHDF | Donor I | LT (Wild Type) | LT_d8_DonorI.fastq.gz |
Wild-Type LT |
| LT_WT_DII | NHDF | Donor II | LT (Wild Type) | LT_d8_DonorII.fastq.gz |
Wild-Type LT |
| LTtr_DI | NHDF | Donor I | LTtr (Truncated) | LTtr_d8_DonorI.fastq.gz |
Tumor-associated truncated form |
| LTtr_DII | NHDF | Donor II | LTtr (Truncated) | LTtr_d8_DonorII.fastq.gz |
Tumor-associated truncated form |
| LT_K331A_DI | NHDF | Donor I | LT-K331A (Mutant) | LT_K331A_d8_DonorI.fastq.gz |
Helicase domain mutant |
| LT_K331A_DII | NHDF | Donor II | LT-K331A (Mutant) | LT_K331A_d8_DonorII.fastq.gz |
Helicase domain mutant |
| LT_K331A_DII_Re | NHDF | Donor II | LT-K331A (Mutant) | LT-K331A-d8-DII_re.fastq.gz |
Replicate or re-test data for Donor II |
Data Analysis Recommendations
-
ChIP-Seq Analysis Strategy:
- Grouping: Divide samples into
LT_only(hTERT_LT_r2, NHDF_Donors),LT+sT(HEK293, hTERT_LT+sT, PFSK), andMock. - Control Handling: Note that PFSK-1A uses IgG controls, while other samples primarily use Input. When calling peaks (e.g., using MACS2), adjust parameters or process them separately based on the control type.
- Integration: Overlap LT binding sites with Differentially Expressed Genes (DEGs) from RNA-Seq. Pay particular attention to whether the LT-K331A mutation leads to the loss or weakening of LT binding at the promoter regions of key genes.
- Grouping: Divide samples into
-
RNA-Seq Analysis Strategy:
- Differential Expression: Use DESeq2 for the following comparisons:
LT_WT vs ControlLTtr vs ControlLT_K331A vs ControlLT_K331A vs LT_WT(Key comparison: Identify genes specifically affected by the K331A mutation)
- Functional Enrichment: Focus on Interferon signaling pathways (ISGs), cell cycle regulation, and DNA damage response genes. According to published literature, LT induces ISGs, and the K331A mutation may alter this induction capacity or affect other downstream pathways.
- Differential Expression: Use DESeq2 for the following comparisons:
-
Multi-Omics Integration:
- Utilize data from NHDF Donor 1 & 2 for cross-omics integration, as these samples have both ChIP-seq (LT binding) and RNA-seq (impact of LT/K331A expression) data from the same primary cell system, offering the highest biological consistency.