β-Thalassemia is one of the most common genetic blood diseases and is caused by either point mutations or deletions in the β-globin (HBB) gene. The generation of patient-specific induced pluripotent stem cells (iPSCs) and subsequent correction of the disease-causing mutations may be a potential therapeutic strategy for this disease. Due to the low efficiency of typical homologous recombination, endonucleases, including TALENs and CRISPR/Cas9, have been widely used to enhance the gene correction efficiency in patient-derived iPSCs. Here, we designed TALENs and CRISPR/Cas9 to directly target the intron2 mutation site IVS2-654 in the globin gene. We observed different frequencies of double-strand breaks (DSBs) at IVS2-654 loci using TALENs and CRISPR/Cas9, and TALENs mediated a higher homologous gene targeting efficiency compared to CRISPR/Cas9 when combined with the piggyBac transposon donor. In addition, more obvious off-target events were observed for CRISPR/Cas9 compared to TALENs. Finally, TALENs-corrected iPSC clones were selected for erythroblast differentiation using the OP9 co-culture system and detected relatively higher transcription of HBB than the uncorrected cells. This comparison of using TALENs or CRISPR/Cas9 to correct specific HBB mutations in patient-derived iPSCs will guide future applications of TALENs- or CRISPR/Cas9-based gene therapies in monogenic diseases.

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BackgroundHuman papillomaviruses (HPVs) are strongly associated with the development of cervical carcinoma, and the distribution of HPV genotypes varies regionally.MethodsTo investigate the distribution characteristics of different genotypes of HPV infection in women in Wuhan, China, a total of 13 775 patients were enrolled over 2 years.ResultsOf these, 2436 patients were infected with HPVs, and the total infection rate was 17.68%. The infection rate of high‐risk HPV (HR‐HPV) was significantly higher (13.96%) than that of single low‐risk HPV (LR‐HPV; 3.72%). Among the HR‐HPV infections, the most common genotype was HPV 52 with an infection rate of 4.23%, followed by HPVs 16, 58, 39, and 51. The most common LR‐HPV genotype was HPV 81, followed by HPVs 6, 11, and 44. Patients under the age of 25 years were found to have the highest HPV infection rate (P < .05). After the age range of 51‐55 years, a downward trend in total HPVs and HR‐HPVs was observed. The HPV infection rate for a single genotype was higher than that for multiple HPVs (P < .01), and the detection rates in summer and winter were significantly higher than those in spring and autumn.ConclusionsThe results demonstrate that the distribution characteristics of various HPV genotype infections are associated with region and age and may be related to season. These data could be the basis for further epidemiological analysis into the control and prevention of HPV infection in this region.