Cloning of Human ABCB11 Gene in E. coli required the removal of an Intragenic Pribnow-Schaller Box before it’s Insertion into Genomic Safe Harbor AAVS1 Site using CRISPR Cas9

Sequencing data and original images including some of the failed experiments can be found here. Context: Please refer the manuscript. Sequencing data: 1) Cloning of ABCB11--sequencing 2) Cloning of gRNA to two Cas9 vectors 3) Sequencing for ABCB11 integration to HEK293T cell line genome at AAVS1 site 4) Sequencing for off target sites-AAVS1 site HEK293T cell genomic DNA PAGE and Western Blot Data: 1) E. Coli lysate transformed ad untransformed DH5a PAGE and W/B with anti ABCB11 antibody 2) ABCB11 transfected cell lines--whole protein fibroblasts 3) ABCB11 integration mediated by Cas9 cell line-whole protein fibroblasts 4) Puromycin selection cellines HEK Agaraose gel images: T7 assay and PCRs Comet Assay

测序数据与原始图像（含部分失败实验的相关内容）可在此处获取。
背景说明：请参阅研究手稿。
测序数据：
1) ABCB11克隆测序
2) 将向导RNA（gRNA）克隆至两种Cas9载体
3) 检测ABCB11整合至HEK293T细胞系基因组AAVS1位点的测序实验
4) 针对HEK293T细胞基因组DNA的AAVS1位点脱靶位点的测序实验
聚丙烯酰胺凝胶电泳（PAGE）与蛋白质免疫印迹（Western Blot, W/B）数据：
1) 转化与未转化DH5α大肠杆菌的裂解液，使用抗ABCB11抗体进行PAGE与W/B检测
2) 转染ABCB11的细胞系——成纤维细胞全蛋白样品
3) Cas9介导ABCB11整合的细胞系——成纤维细胞全蛋白样品
4) 嘌呤霉素筛选的HEK293T细胞系
琼脂糖凝胶电泳图像：T7酶切实验（T7 assay）与聚合酶链式反应（PCR）扩增产物
彗星实验（Comet Assay）