Disruptive Technology: CRISPR/Cas-Based Tools and Approaches

Abstract

Designer nucleases are versatile tools for genome modification and therapy development and have gained widespread accessibility with the advent of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) technology. Prokaryotic RNA-guided nucleases of CRISPR/Cas type, since first being adopted as editing tools in eukaryotic cells, have experienced rapid uptake and development. Diverse modes of delivery by viral and non-viral vectors and ongoing discovery and engineering of new CRISPR/Cas-type tools with alternative target site requirements, cleavage patterns and DNA- or RNA-specific action continue to expand the versatility of this family of nucleases. CRISPR/Cas-based molecules may also act without double-strand breaks as DNA base editors or even without single-stranded cleavage, be it as epigenetic regulators, transcription factors or RNA base editors, with further scope for discovery and development. For many potential therapeutic applications of CRISPR/Cas-type molecules and their derivatives, efficiencies still need to be improved and safety issues addressed, including those of preexisting immunity against Cas molecules, off-target activity and recombination and sequence alterations relating to double-strand-break events. This review gives a concise overview of current CRISPR/Cas tools, applications, concerns and trends.

The global preclinical antibacterial pipeline

“…New genetic tools such as CRISPR-Cas systems are used to genetically engineer phages that infect diverse hosts 46 . Phages are also used as species-specific carriers for a variety of potential antibacterial payloads 47 or CRISPR-Cas-based RNA-guided nucleases targeted at resistance or virulence determinants 48,49 . Considerable progress has been made recently in tackling the great challenges in the chemistry, manufacturing and control of therapeutic phages, especially in production, stability, purity and quality control.…”

Mapping the coevolution, leadership and financing of research on viral vectors, RNAi, CRISPR/Cas9 and other genomic editing technologies

“…These three genome editing platforms are considered not just precursors of CRISPR/Cas9 [8] but also incumbent technologies that eventually would be supplanted by CRISPR/Cas9 [9]. Interestingly, viral vectors and RNAi have been also considered incumbent genetic therapeutic technologies with reference to CRISPR/Cas9 [10] which is considered a disruptive innovation [11]. As a matter of fact, the relation between viral vectors, RNAi and CRISPR/Cas9 is more sophisticated.…”

“…Comparing the trajectories of these six technologies (Viral vectors, RNAi and the four genome editing platforms) as well as analyzing the interactions that take place between them is relevant for the general understanding of the current processes of technological invention and development that take place in the biotechnological field. This is due to the different times in which these biotechnologies emerged [1][2][3][4][5][6][7], the revolutionary and innovative character attributed to them [1][2][3][4][5][6][7] and the competitive and collaborative relationships that exist between these technologies [8][9][10][11][12][13][14]. A first step to analyse the coevolution of these technologies is by the identification and analysis of research fronts (clusters of papers in the citation networks).…”

Theranostics of Genetic Diseases

“…For this reason, the term 'theranostics' has been proposed-combining 'therapeutic strategies' and 'diagnostics' within the same term. Four articles in this issue review therapeutic approaches, discussing the issue of gene therapy [1], gene editing [2,3], and micro-RNA (miRNA) therapeutics [4]. Ghiaccio et al [1] review gene therapy for β-thalassemia with respect to the milestones reached, novel developments, and present (and future) challenges.…”

“…The relatively new approach based on gene editing is the object of two review articles by Lederer and Kleanthous [2,3]. CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9-based tools and therapy development for rare genetic diseases is indeed a fascinating possibility for site-specific correction of the genetic mutations causing hereditary pathologies, as also summarized in other review articles available in the recent literature [9][10][11][12][13].…”