br Author contributions br Resource table Resource details

Author contributions
Resource table. Resource details Previously we have generated an induced pluripotent stem cell line (named MHHi002-A) from a male CF patient homozygous for the mutation p.Phe508del in the CFTR gene. Thereby the absence of the three bases CTT results in the deletion of phenylalanine (Phe) in position 508 of the amino buy latrunculin a sequence (Fig. 1A). For the generation of patient-specific iPSCs, endothelial colony forming cells (ECFCs) from the peripheral blood of the CF patient were reprogrammed through overexpression of the codon optimized human pluripotency factors OCT4, SOX2, KLF4 and c-MYC using a 4-in-1 lentiviral vector (Warlich et al., 2011). Here, we used this CF-iPSC line for the correction of the mutated triplet CTT by TALENs and single-stranded oligonucleotides (ssODNs) by applying our established targeting protocol (Merkert et al., 2014). In detail, TALEN plasmids and ssODN were transfected into MHHi002-A cells and by limiting dilution and PCR screening we identified 5 heterozygously targeted clones without any pre-selection. One clone was chosen for further characterization (MHHi002-A-1). Sequence analysis confirmed the mono-allelic correction of the p.Phe508del mutation and karyotype analysis proved chromosomal integrity (Fig. 1C, D). The corrected CF-iPSC line MHHi002-A-1 expresses endogenous core pluripotency markers like OCT4, NANOG, SSEA-4 and TRA-1-60 and could be differentiated in vitro into derivatives of all three germ layers (Fig. 1E, F).
Materials and methods
Resource table Resource utility
Resource details This study was approved by the ethics committee of the 1st Affiliated hospital of Zhengzhou University. Written informed consent was acquired from the patient. The diagnosis of PD was made following the UK Brain Bank criteria. This 65year old patient has bradykinesia and muscular rigidity, and he has no other features suggesting a different condition, and his Parkinsonism improves with dopaminergic medication. Skin fibroblasts were collected from the skin biopsy of the patient. The messenger RNA reprogramming kit (Stemgent, USA) used to derive iPSC included 5 reprogramming factors: Klf-4, c-Myc, Oct-4, Lin-28, and Sox-2. This reprogramming protocol ensures the successful establishment of transgene-free iPSC lines. The derived iPSC colonies with typical embryonic stem cell morphology were picked and expanded (Fig. 1B, 1C). The expression of pluripotent markers in the iPSC was confirmed by immunofluorescence staining (Fig. 1E). The iPSC line could form embryoid bodies in vitro (Fig. 1D), and this line could also form teratoma in vivo (Fig. 1G). The derived hiPSC line also showed normal karyotype (46, XY, Fig. 1H).
Materials and methods
Acknowledgments This study was supported by a grant from National Natural Science Foundation of China (NSFC, project 81570199).
Resource Table
Resource utility KLF15 is required for normal heart homeostasis and becomes downregulated upon cardiac hypertrophy in mice and human patients (Fisch et al., 2007; Noack et al., 2012). The HES-KLF15-KO line was generated to study the pathological mechanisms triggered by KLF15 deletion in human cardiomyocyte 2D and 3D in vitro models.
Resource details The transcription factor KLF15 regulates diverse cellular processes and is mainly expressed in liver, kidney, skeletal and cardiac muscle. Particularly in the adult heart it is essential to maintain normal cardiac function (Fisch et al., 2007; Noack et al., 2012) and regulates cell proliferation, hypertrophy, and lipid metabolism (Haldar et al., 2012). Importantly, KLF15 is downregulated in human and rodent cardiomyopathies, suggesting an evolutionary conserved mechanism. To explore its role in human cardiomyocytes, we deleted KLF15 function by generating a homozygous hESC KLF15-KO line. We used a double-paired CRISPR/Cas9-D10A nickase (Cas9n) gene editing approach to minimize potential off-target effects. To target the genomic locus of KLF15, we designed four sgRNAs with a cutting offset of 128bp including the splice acceptor and the transcription start site (TSS) (Fig. 1A). HES2 cells were nucleofected with all four sgRNAs expressing plasmids, also containing Cas9n and puromycin resistance. Cells were selected with puromycin and seeded at very low density to allow colony growth from single cells. After PCR genotyping (Fig. 1B, primer sites shown in Fig. 1A) and sequencing, positive colonies (KLF15-KO) were expanded and cryopreserved. Nucleofected cells having wild-type KLF15 (KLF15-WT) locus served as controls. The lines were authenticated by STR analysis and tested negative for mycoplasma.