CRISPR-Cas systems for editing, regulating and targeting genomes

Abstract

Targeted genome editing using engineered nucleases has rapidly transformed from a niche technology to a mainstream method used by many biological researchers. This widespread adoption has been largely fueled by the emergence of the clustered regularly interspaced short palindromic repeat (CRISPR) technology, an important new platform for generating RNA-guided nucleases (RGNs), such as Cas9, with customizable specificities. RGN-mediated genome editing is facile, rapid and has enabled the efficient modification of endogenous genes in a wide variety of biomedically important cell types and novel organisms that have traditionally been challenging to manipulate genetically. Furthermore, a modified version of the CRISPR-Cas9 system has been developed to recruit heterologous domains that can regulate endogenous gene expression or label specific genomic loci in living cells. Although the genome-wide specificities of CRISPR-Cas9 systems remain to be fully defined, the capabilities of these systems to perform targeted, highly efficient alterations of genome sequence and gene expression will undoubtedly transform biological research and spur the development of novel molecular therapeutics for human disease.

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

“…Broader studies are still required to pinpoint the contribution of donor genetic background and subsequent epigenetic modifications on both cardiac phenotype and drug responses. This could be provided by recent advances in genome‐editing techniques, with CRISPR/Cas9 being the current leading method (Sander and Joung, 2014). This approach allows rapid and precise modification of the hPSC genome so that generating isogenically matched controls for (genetically) diseased hiPSC lines is feasible for many labs, reducing one source of variability.…”

State of the art technologies to explore long non‐coding RNAs in cancer

“…Making use of two gRNAs, the dual CRISPR system is suited for perturbing lncRNAs in contrast to conventional methods, which may lead to insertions or deletions, not enough to eliminate the function of lncRNAs 70, 72 (Table 1). …”

The Flipside of the Power of Engineered T Cells: Observed and Potential Toxicities of Genetically Modified T Cells as Therapy

“…44 More recently, clustered regularly-interspaced short palindromic repeats (CRISPR)-based gene delivery approaches have demonstrated promising results with the possibility of non-viral targeting transgene insertion. 45 Various transposase-based systems have now entered clinical trials, but their overall safety profile has yet to be fully evaluated. Transient mRNA expression seems appealing from a safety perspective but lacks the long-term expression needed for maximal CAR T cell function and persistence.…”