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CRISPR to fix bad blood: a new tool in basic and clinical hematology
Ferrata Storti Foundation
Haematologica 2019 Volume 104(5):881-893
Elisa González-Romero,1 Cristina Martínez-Valiente,1 Cristian García-Ruiz,1 Rafael P. Vázquez-Manrique,2,3 José Cervera4,5 and Alejandra Sanjuan-Pla1
1Hematology Research Group, Instituto de Investigación Sanitaria La Fe, Valencia; 2Grupo de Investigación en Biomedicina Molecular, Celular y Genómica, Instituto de Investigación Sanitaria La Fe, Valencia; 3CIBER de Enfermedades Raras, Madrid; 4Hematology Department, Hospital Universitari i Politècnic La Fe, Valencia and 5CIBER de Oncología, Madrid, Spain
ABSTRACT
Advances in genome engineering in the last decade, particularly in the development of programmable nucleases, have made it possi- ble to edit the genomes of most cell types precisely and efficiently. Chief among these advances, the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system is a novel, versatile and easy- to-use tool to edit genomes irrespective of their complexity, with multi- ple and broad applications in biomedicine. In this review, we focus on the use of CRISPR/Cas9 genome editing in the context of hematologic dis- eases and appraise the major achievements and challenges in this rapidly moving field to gain a clearer perspective on the potential of this technol- ogy to move from the laboratory to the clinic. Accordingly, we discuss data from studies editing hematopoietic cells to understand and model blood diseases, and to develop novel therapies for hematologic malignan- cies. We provide an overview of the applications of gene editing in exper- imental, preclinical and clinical hematology including interrogation of gene function, target identification and drug discovery and chimeric anti- gen receptor T-cell engineering. We also highlight current limitations of CRISPR/Cas9 and the possible strategies to overcome them. Finally, we consider what advances in CRISPR/Cas9 are needed to move the hema- tology field forward.
Introduction
Genome engineering is defined as the deliberate modification of an organism’s genetic material. It has been used since the early 1980s to study the impact of DNA mutations in human disease precisely and has helped to unravel the genetic basis of many malignancies and to advance their diagnosis, prevention, and treatment. Genome engineering to introduce defined alterations has traditionally employed homologous recombination strategies to modify a gene of interest (gain- or loss-of- function) using segments of exogenous DNA.1 To achieve homologous recombina- tion in the “pre-nuclease” era, large DNA sequences homologous to the target sequence, containing sequence changes designed to produce the desired modifica- tion, were introduced into the nucleus of the receiving cell. This technology depends heavily on chance since the DNA construct is expected to interact with the target and induce gene conversion upon recombination of DNA homology arms. The success rate of this technology was historically extremely low, which together with the complexity in designing targeting vectors, and the time and resources required, put it out of reach of some researchers. However, with the advent of highly-specific chimeric nucleases (which are able to recognize 18 or more base pairs) to induce locus-specific DNA double-strand breaks (DSB), the effi- ciency of homologous recombination rose substantially (e.g., becoming more than 40,000 times more efficient2), depending on the experimental system. The use of such nucleases has increased in recent years with the development of meganucle- ases,3 zinc-finger nucleases,4 and transcription activator-like effector nucleases
Correspondence:
ALEJANDRA SANJUAN PLA
alejandra_sanjuan@iislafe.es
Received: December 20, 2018. Accepted: February 19, 2019. Pre-published: March 28, 2019.
doi:10.3324/haematol.2018.211359
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/104/5/881
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