Improving the Function of Crispr-Cas9 for Genome Editing Therapy: Editing the Editor

Abstract

ABSTRAK Kata kunci: CRISPR, Cas9, efektivitas, spesifisitas, terapi gen ABSTRACTGene editing has become reasonably easy since the discovery of clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9). Most genetic diseases cannot be treated causally, and currently available therapies are mainly symptom-based. To treat the etiology of genetic diseases, a firm gene editing therapy is necessary to be established. This posits Cas9-facilitated gene editing as a prospective modality to become a clinically approved therapy in the future to treat genetic disorders. However, until recently, Cas9-based genome editing is still facing several hurdles, including low specificity, low effectiveness, and difficult delivery. Currently available Cas9 nucleases are able to bind to non-specific DNA sequence and produce non-specific cleavage. The efficiency has been relatively low due to the preference of non-homologous end-joining (NHEJ) over homology-directed repair (HDR) by the host cell. Furthermore, in order to deliver Cas9 into the nucleus, multiple physiological barriers have to be overcome. This review discussed recent developments in tackling these three hurdles, ranging from designing the guide RNA using multiple bioinformatics tools, modifying Cas9 structure, as well as packaging the nuclease-guide RNA complex into viral vectors and nanoparticles. Considering the active research on this area, it is expected that CRISPR/Cas9 can be utilized as a clinical therapy in the near future.

THE DEVELOPMENT of a SAFE and OPTIMIZED GENE THERAPY for HUMAN DISEASES

“…This review does not cover extensively the successful clinical trials for sickle cell anemia, thalassemia major, lymphoma (Kumar, Markusic, Biswas, High, & Herzog, 2016), nor the potential future for genome-editing therapy (Gabriel, von Kalle, & Schmidt, 2015;Supit, 2017) which will broaden the scope of gene therapy for the benefits of humanity. As more genes are being identified and their ontologies are being refined, future direction of gene therapy would be more interesting, sophisticated, yet euphorically challenging.…”