iMEF SDHC-loss stable line ATAC-seq. iMEF SDHC-loss stable line ATAC-seq

We have developed a tet-inducible SV-40 large T antigen-expressing lentivirus-immortalized mouse embryonic fibroblast (iMEF) cell culture model of mitochondrial electron transport chain (ETC) dysfunction involving complex II (succinate dehydrogenase; SDH) in which silencing gene rearrangement of the Sdhc floxed allele is driven by doxycycline-dependent expression of cre-recombinase from a tet-inducible promoter (R26M2rtTA/+;TetOcre;Sdhcfl/fl). For comparison, we include an isogenic Sdhc wt control line (R26M2rtTA/+;TetOcre;Sdhcfl/wt) that retains one intact copy of Sdhc upon doxycycline exposure. Following doxycycline induction of both cell lines, dilutional subcloning was employed to obtain stable Sdhc -/- (SDHC KO) and Sdhc +/- (Control) cell lines. Sdhc gene rearrangement status for each clonally-derived cell line was confirmed by PCR. Cell lines were grown in standard DMEM containing penicillin/streptomycin antibiotics (0.5 mg/mL), non-essential amino acids (100 micromolar each of glycine, alanine, asparagine, aspartic acid, glutamic acid, proline, and serine), sodium pyruvate (1 mM), and HEPES buffer (10 mM) at 21% O2 and 5% CO2. Following cell line derivation and verification of Sdhc genetic status, accessible chromatin regions in each cell line were interrogated by ATAC-seq. Overall design: Included in this dataset are single replicate ATAC-seq data for two SDHC KO (R26M2rtTA/+;TetOcre;Sdhc-/-) and two control (R26M2rtTA/+;TetOcre;Sdhc+/-) cell lines derived by dilutional subcloning following induction of Sdhc gene rearrangement with doxycycline.

本研究构建了四环素诱导型、表达猿猴病毒40（SV-40）大T抗原的慢病毒永生化小鼠胚胎成纤维细胞（iMEF）模型，用于模拟线粒体电子传递链（ETC）复合物II（琥珀酸脱氢酶，SDH）功能异常。该模型中，Sdhc floxed等位基因的基因重排由四环素诱导型启动子（R26M2rtTA/+;TetOcre;Sdhcfl/fl）驱动的Cre重组酶表达所介导。
为进行对照实验，本研究同时设置同基因背景的Sdhc野生型对照细胞系（R26M2rtTA/+;TetOcre;Sdhcfl/wt），该细胞系在多西环素处理后仍保留一个完整的Sdhc等位基因。
对两种细胞系施加多西环素诱导后，采用有限稀释亚克隆法获得稳定的Sdhc纯合敲除（Sdhc-/-，SDHC KO）与Sdhc杂合（Sdhc+/-，对照）细胞系；通过聚合酶链式反应（PCR）验证各克隆细胞系的Sdhc基因重排状态。
细胞系采用含青霉素/链霉素（0.5 mg/mL）、非必需氨基酸（每种氨基酸终浓度为100 μM，包括甘氨酸、丙氨酸、天冬酰胺、天冬氨酸、谷氨酸、脯氨酸与丝氨酸）、丙酮酸钠（1 mM）及羟乙基哌嗪乙硫磺酸（HEPES）缓冲液（10 mM）的标准达尔伯克改良伊格尔培养基（DMEM）进行培养，培养条件为21% O2、5% CO2。
完成细胞系构建与Sdhc遗传状态验证后，采用转座酶可及性测序（ATAC-seq）检测各细胞系的染色质开放区域。
数据集整体设计如下：包含经多西环素诱导Sdhc基因重排后，通过有限稀释亚克隆获得的2株SDHC纯合敲除细胞系（R26M2rtTA/+;TetOcre;Sdhc-/-）与2株对照细胞系（R26M2rtTA/+;TetOcre;Sdhc+/-）的单重复ATAC-seq数据。