Abstract: Methods are provided herein for assembling at least two nucleic acids using a sequence specific nuclease agent (e.g., a gRNA-Cas complex) to create end sequences having complementarity and subsequently assembling the overlapping complementary sequences. The nuclease agent (e.g., a gRNA-Cas complex) can create double strand breaks in dsDNA in order to create overlapping end sequences or can create nicks on each strand to produce complementary overhanging end sequences. Assembly using the method described herein can assemble any nucleic acids having overlapping sequences or can use a joiner oligo to assemble sequences without complementary ends.

Abstract: The invention provides a method of inducing ?-cell neogenesis from epithelial cells. The method includes the step of exposing epithelial cells that have a disrupted G1-S cell cycle transition to a hedgehog protein in an amount effective to stimulate ?-cell neogenesis from the epithelial cells. Cells resulting from the method are insulin-positive and express pancreatic progenitor cell markers including Pdx-1 and ngn3. ?-cell population expansion is observed when the method is carried out in either in vitro or in vivo settings.