TY  - GEN
T1  - Engineering essential genes with a “jump board” strategy using CRISPR/Cas9
AU  - Duan, Ye
AU  - Choi, Sungwook
AU  - Nelson, Charles
AU  - Ambros, Victor
DO  - 10.17912/micropub.biology.000315
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000315/
AB  - The application of CRISPR/Cas9 genome editing has revolutionized the genetic analysis of gene function in vivo by enabling highly efficient and precise in loco mutagenesis of defined genomic loci (Adli, 2018). In particular, CRISPR/Cas9 mediated homologous directed recombination (HDR) permits the introduction of complex genetic modifications at single nucleotide resolution (Cong et al., 2013; Dickinson et al., 2013). However, the targeted mutagenesis of essential genes using CRISPR/Cas9 can be challenging since the recovery of deleterious mutations of an essential gene may require a genetic balancer with a wild type copy of the gene (or alternatively a genetic suppressor locus) that can be later outcrossed for phenotypic analysis of novel targeted mutations. Genetic balancers are commonly available for model organisms such as worms or flies, but in the context of CRISPR/Cas9 genome editing, the balancer itself can be edited along with the targeted genomic locus, dramatically reduce the efficiency of recovering the desired loss of function alleles. Therefore, editing of essential genes would be facilitated by a strategy where the targeted genomic locus is efficiently edited, whilst the balancer locus is resistant to editing.
PY  - 2020
JO  - microPublication Biology
ER  -