Page 38 - 2019_05-HaematologicaMondo-web

E. González-Romero et al.
evaluate therapeutic CRISPR/Cas9 gene editing to correct mutations in pathologies such as autoimmune diseases and multiple myeloma.
Given the exponential output of scientific publications using CRISPR in the last years, there is a clear need to syn- thesize the latest data on gene editing. With this in mind, here we describe recent advances in the use of CRISPR/Cas9 in hematologic research and clinical transla- tion. We also consider the limitations of CRISPR/Cas9 technology for therapeutic applications, their possible solutions, and how the field of hematology may move for- ward. While CRISPR/Cas9 gene editing is hoped to be a treatment for many hematologic diseases, large clinical tri- als are needed to evaluate the efficacy and safety of CRISPR/Cas9 for patients, a promising area that will undoubtedly expand in the near future.
A
CRISPR/Cas9 in hematology research
In vitro gene editing of blood cells
Hematopoietic cell lines are a robust model for validating
gRNA specificity and CRISPR/Cas9 experimental design because of their easy manipulation and expansion, and enrichment of edited cells. In this respect, cell lines have been employed: (i) to analyze gene function upon NHEJ- mediated gene disruption; (ii) to insert a point mutation or a DNA fragment; (iii) to correct a point mutation, and (iv) to create chromosomal translocations. Although valuable as a proof-of-concept approach, editing success cannot always be extrapolated to difficult-to-edit primary cells.
Many studies have reported successful CRISPR/Cas9- mediated gene editing in hematopoietic cell lines. As a consequence of indels introduced by NHEJ-mediated DSB
B
Figure 2. CRISPR/Cas9-mediated genome editing in hematology. (A) Illustration of the CRISPR/Cas9 sys- tem. Site-specific DSB are produced by CRISPR/Cas9 and are either repaired by NHEJ, introducing indels that provoke gene disruption, or by HDR that, in the presence of a DNA
template, creates translocations, or point mutations. gRNA: guide RNA; DSB: double- strand break; NHEJ: non-homolo- gous end joining; HDR: homology- directed repair; indel: insertions and deletions. (B) Applications of CRISPR/Cas9 technology in hema- tology research and human therapy. HIV: human immunodeficiency virus; CAR: chimeric antigen recep- tor; CHIP: clonal hematopoiesis of indeterminate potential.
insertions,
884
haematologica | 2019; 104(5)