Zebrafish Genome Engineering Using the CRISPR-Cas9 System.
Geneticists have long sought the ability to manipulate vertebrate genomes by directly altering the information encoded in specific genes. The recently discovered clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 endonuclease has the ability to bind single loci within vertebrate genomes and generate double-strand breaks (DSBs) at those sites. These DSBs induce an endogenous DSB repair response that results in small insertions or deletions at the targeted site. Alternatively, a template can be supplied, in which case homology-directed repair results in the generation of engineered alleles at the break site. These changes alter the function of the targeted gene facilitating the analysis of gene function. This tool has been widely adopted in the zebrafish model; we discuss the development of this system in the zebrafish and how it can be manipulated to facilitate genome engineering.
School of Pharmaceutical Sciences, Xiamen University, Xiamen 361102, China.
Mingyu
M
Li
Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China.
Liyuan
L
Zhao
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Division of Nephrology and Hypertension, Vanderbilt Center for Kidney Disease, Vanderbilt University Medical Center, Nashville, TN. Electronic address: patrick.s.page-mccaw@vanderbilt.edu.
Patrick S
PS
Page-McCaw
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232, USA. Electronic address: wenbiao.chen@vanderbilt.edu.
Wenbiao
W
Chen
D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D053903:DNA Breaks, Double-Stranded; D017353:Gene Deletion; D005818:Genetic Engineering; D016678:Genome; D016254:Mutagenesis, Insertional; D015027:Zebrafish / Q000235:genetics*
CRISPR–Cas9; genetic screen; genome editing; knock-in; knockout; zebrafish