IASO: Precise genome editing idealizing backbone healing and tale-guided piggyBac

Abstract

Gene editing, which is the alteration of the genetic material of a living organism by inserting, replacing or deleting a DNA sequence, could provide a long-term solution for patients suffering from genetic diseases. A major challenge in the field is the efficient and precise insertion of DNA sequences without off-target insertions. While CRISPR/Cas9 has revolutionized genome engineering, its clinical applicability is limited due to potential off-target effects, limited efficiency in dividing cells, and having a limited DNA cargo capacity. Thus, this thesis aims to optimize a novel approach that would result in a specific and efficient method of transferring a gene of interest to a particular target in the genome. We hypothesize that the piggyBac transposon (pB), TALE and non-homologous end joining (NHEJ) can be idealized to target and integrate at a specific DNA target site.