DECKO: Single-oligo, dual-CRISPR deletion of genomic elements including long non-coding RNAs
Recently, Estel Aparicio-Prat et al. published a study about a new genome editing system based on CRISPR and lentiviral vector technologies, this tool was specifically designed to knockout long non-coding RNAs. They spell this new system DECKO for Double Excision CRISPR Knockout. The key feature of DECKO is the used of e lentiviral vectors expressing two guide RNAs (gRNAs) simultaneously. The scientists tested this tool to delete fragments ranging in size from 100 to 3000bp in UCA1, TFRC and MALAT1 genes. They obtain the best results with MALTA1 gene with which they managed to derive 9 homozygous knockout clones, representing 4 distinct CRISPR targeting constructs in 3 different cell lines. These homozygous clones were derived at a rate of between 1/ 26 and 4/15 clones. These knockout clones had the desired phenotypic effects: RNA levels were reduced to between 2 and 41 % of control levels, resulting in a reduction of cellular proliferation, a key MALAT1 phenotype.
Although a number of technical features remain to improve, the authors give an interesting proof of concept about a practical way of knocking out lncRNAs. One can imagine that future works will be published about the improvement and the use in other context of this system.