Development and Applications of CRISPR-Cas9 for Genome Engineering

By P. Hsu, Eric S. Lander, Feng Zhang. 2014

5,034 citations recorded by the source corpus.

Recent advances in genome engineering technologies based on the CRISPR-associated RNA-guided endonuclease Cas9 are enabling the systematic interrogation of mammalian genome function. Analogous to the search function in modern word processors, Cas9 can be guided to specific locations within complex genomes by a short RNA search string. Using this system, DNA sequences within the endogenous genome and their functional outputs are now easily edited or modulated in virtually any organism of choice. Cas9-mediated genetic perturbation is simple and scalable, empowering researchers to elucidate the functional organization of the genome at the systems level and establish causal linkages between genetic variations and biological phenotypes. In this Review, we describe the development and applications of Cas9 for a variety of research or translational applications while highlighting challenges as well as future directions. Derived from a remarkable microbial defense system, Cas9 is driving innovative applications from basic biology to biotechnology and medicine.

Subfields: Genetics, Molecular Biology

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Citations in the corpus

The potential for the use of gene drives for pest control in New Zealand: a perspective (2018, background): “The advent of CRISPR/Cas9 targeting technologies (Hsu et al. 2014) has given new life to the gene drive idea.”
CRISPR-Cas9 Knockin Mice for Genome Editing and Cancer Modeling (2014, background): “…of Health Sciences and Technology 8Institute for Medical Engineering and Science 9Department of Chemical Engineering Massachusetts Institute of Technology, Cambridge, MA 02139, USA 10Harvard Medical School, Boston, MA 02115, USA 11Center for Immunology and Inflammatory Diseases,…”
CRISPR/Cas9 in Genome Editing and Beyond. (2016, methodology): “The Cas9-mediated gene-editing system has been broadly used in reverse genetics studies to understand the role of specific genes, for disease modeling, and for demonstrating new therapeutic schemes in a number of models of genetic and infectious diseases (9, 10, 94).”
Transcriptome engineering with RNA-targeting Type VI-D CRISPR effectors (2018, neutral): “1,400 aa depending on subtype, with compact outliers such as CjCas9 or SaCas9), Cas13a (1,250 aa), Cas13b (1,150 aa), and Cas13c (1,120 aa) (Figure S2B) (Chylinski et al., 2013; Cox et al., 2017; Hsu et al., 2014; Kim et al., 2017; Shmakov et al., 2015; Smargon et al., 2017).”
In vivo genome editing using Staphylococcus aureus Cas9 (2015, background): “Analysis of over 600 Cas9 orthologs shows that these enzymes are clustered into two length groups with characteristic protein sizes of approximately 1350aa and 1000aa residues, respectively 19,21 (Extended Data Fig.”