CRISPR-Cas9 heterodimers fosters precise chromosomal deletions in human cells

Genome editing based on dual CRISPR-Cas9 complexes (multiplexes) permits removing specific genomic sequences in living cells and leverages research on functional genomics and genetic therapies. Delivering the required large and multicomponent reagents in a synchronous and stoichiometric manner remain challenging. Moreover, uncoordinated activity of independently acting CRISPR-Cas9 multiplexes increases the complexity of genome editing outcomes. We investigate the potential of fostering precise multiplexing genome editing through synchronous deliverying of Cas9 ortholog fusion constructs (forced Cas9 heterodimers) alone or together with their cognate guide RNAs (forced CRISPR-Cas9 heterodimers) using high-capacity adenovector particles (AdVPs).