Additional file 1: of Application of long single-stranded DNA donors in genome editing: generation and validation of mouse mutants

Table S1. Sequences of reagents used in the study. The table shows the sequences of the oligonucleotides and lssDNA donors, primers and TaqMan assays employed in this study. LoxP sites (for all conditional projects) and point mutations (for Gckr and Rims1 project) are underlined. Sequences added for diagnostic (for all conditional projects except Syt7) and silent mutations (for Gckr and Rims1 project) are shown in italics. For the plasmids, sequences flanked by and including homology arms are shown. The ddPCR reference copy counting assay is labelled with VIC. All other ddPCR copy counting assays are labelled with fluorescein amidite (FAM). Copy counting assays labelled as UNIV ddPCR assays recognize both WT and engineered alleles; MUT ddPCR assays recognize engineered allele only. Table S2. Production of founders for conditional alleles. The table shows the numbers of embryos and animals involved in mutagenesis attempts employing the injection of CRISPR/Cas9 reagents and lssDNA donors. Table S3. Generation of conditional alleles employing different donor types. The table shows the numbers of embryos and animals involved in mutagenesis attempts employing the injection of CRISPR/Cas9 reagents and oligonucleotides, plasmids or lssDNA donors. The results of the analysis of the founders obtained from these attempts are also summarized. Table S4. Generation of a Rims1R655H point mutation. Further genotype screening data for this project are shown in Additional file 18: Figure S16 and Additional file 19: Figure S17. Table S5. Analysis of the Rims1R655H project. The table details the results of screening of five positive F0 animals obtained for the generation of a Rims1R655H point mutation and the subsequent characterization of the F1 animals obtained from mating of these F0 animals to WT mice. Table S6. Nomenclature of new mouse lines established in the course of the study. (XLS 81 kb)

补充表S1. 本研究所用试剂的序列信息。本表格列出了本研究中使用的寡核苷酸（oligonucleotides）、长单链DNA（lssDNA）供体、引物以及TaqMan检测体系（TaqMan assays）的序列信息。本研究所有条件性项目所用的LoxP位点（LoxP sites），以及Gckr与Rims1项目所用的点突变均已添加下划线。为诊断（除Syt7外的所有条件性项目）及沉默突变（Gckr和Rims1项目）所添加的序列以斜体显示。对于质粒，已列出同源臂（homology arms）侧翼且包含同源臂的序列。数字液滴PCR（ddPCR）参考拷贝计数检测以VIC荧光标记。其余所有ddPCR拷贝计数检测均以荧光素酰胺（FAM）标记。标注为UNIV ddPCR检测的拷贝计数检测可识别野生型（WT）与工程化等位基因；标注为MUT ddPCR检测的拷贝计数检测仅可识别工程化等位基因。
补充表S2. 条件性等位基因首建动物的构建。本表格列出了通过注射CRISPR/Cas9试剂与lssDNA供体进行诱变尝试所涉及的胚胎与实验动物数量。
补充表S3. 采用不同供体类型构建条件性等位基因。本表格列出了通过注射CRISPR/Cas9试剂与寡核苷酸、质粒或lssDNA供体进行诱变尝试所涉及的胚胎与实验动物数量，同时总结了上述诱变尝试获得的首建动物的分析结果。
补充表S4. Rims1R655H点突变的构建。本项目的额外基因型筛选数据详见补充文件18的图S16与补充文件19的图S17。
补充表S5. Rims1R655H项目的分析。本表格详细描述了Rims1R655H点突变构建过程中获得的5只阳性F0代动物的筛选结果，以及将这些F0代动物与野生型小鼠交配后获得的F1代动物的后续鉴定结果。
补充表S6. 本研究构建的新型小鼠品系的命名规则（XLS 81 kb）