Next-generation CRISPR gene-drive systems using Cas12a nuclease

Abstract

One method for reducing the impact of vector-borne diseases is through the use of CRISPR-based gene drives, which manipulate insect populations due to their ability to rapidly propagate desired genetic traits into a target population. However, all current gene drives employ a Cas9 nuclease that is constitutively active, impeding our control over their propagation abilities and limiting the generation of novel gene drive arrangements. Yet, other nucleases such as the temperature-sensitive Cas12a have not been explored for gene drive designs. To address this, we herein present a proof-of-concept gene-drive system driven by Cas12a that can be regulated via temperature modulation. Furthermore, we combined Cas9 and Cas12a to build double gene drives capable of simultaneously spreading two independent engineered alleles. The development of Cas12a-mediated gene drives provides an innovative option for designing next-generation vector control strategies to combat disease vectors and agricultural pests.

Manipulating the Destiny of Wild Populations Using CRISPR

“…Initial work utilized Cas9 from Streptococcus pyogenes (121), simply because it was the first nuclease characterized. However, hundreds of new RNA-guided nucleases have been identified, and which type will ultimately prove most robust in specific genomic and physical environments remains to be determined (122,136). As noted above, a typical autonomous HCGD encodes both Cas9 and the gRNA linked at the same loci (Figure 1).…”