CRISPR/Cas9 is a versatile genome-editing technology that is widely used for studying the functionality of genetic elements, creating genetically modified organisms as well as preclinical research of genetic disorders. However, the high frequency of off-target activity (≥50%)—RGEN (RNA-guided endonuclease)-induced mutations at sites other than the intended on-target site—is one major concern, especially for therapeutic and clinical applications. Here, we review the basic mechanisms underlying off-target cutting in the CRISPR/Cas9 system, methods for detecting off-target mutations, and strategies for minimizing off-target cleavage. The improvement off-target specificity in the CRISPR/Cas9 system will provide solid genotype–phenotype correlations, and thus enable faithful interpretation of genome-editing data, which will certainly facilitate the basic and clinical application of this technology.

Key Points• There is a need to identify the best HLA haplotypemismatched related donor.• Use of young, male, NIMAmismatched donors results in the best survival after HLA haplotype-mismatched related donor transplants. Older sister donors were inferior to father donors with regard to NRM (HR 5 1.87; 95% CI 5 1.10-3.20; P 5 .02) and survival (HR 5 1.59; 95% CI 5 1.05-2.40; P 5 .03). Noninherited maternal antigen-mismatched sibling donors were associated with the lowest incidence of acute GVHD compared with parental donors and noninherited paternal antigen-mismatched sibling donors. Specific HLA disparities were not significantly correlated with transplant outcomes. Our data indicate which HLA haplotypemismatched related donors are associated with the best transplant outcomes in persons with hematological neoplasms. (Blood. 2014; 124(6):843-850)

Among HID and ISD recipients, the 3-year disease-free survival rate was 74% and 78% (P 5 .34), respectively; the overall survival rate was 79% and 82% (P 5 .36), respectively; cumulative incidences of relapse were 15% and 15% (P 5 .98); and those of the nonrelapse-mortality were 13% and 8% (P 5 .13), respectively. In conclusion, unmanipulated haploidentical HSCT achieves outcomes similar to those of ISD HSCT for AML patients in CR1. Such transplantation was demonstrated to be a valid alternative as postremission treatment of intermediateor high-risk AML patients in CR1 lacking an identical donor. This trial was registered at www.chictr.org as #ChiCTR-OCH-10000940. (Blood. 2015;125(25):3956-3962)

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