Abstract: Systems and methods for image-driven cell manufacturing are provided. Systems comprise a substrate for cells, imaging system for imaging cells on the substrate, computing system that computes cell characteristics from images, and a pulsed laser scanning system. The substrate is suitable for high-resolution cell imaging and is coated with a layer that partially absorbs laser pulses for the purpose of converting energy into microbubble formation. The computing system communicatively coupled to the pulsed laser scanning system and directs laser pulses to the substrate under targeted cells. Laser pulses are converted into mechanical energy via microbubbles that, depending on laser energy and pulse pattern, destroy selected cells, remove selected cells, or temporarily porate selected cells for the purpose of introducing biological cargos.

Abstract: Methods for identifying targets involved in cell differentiation, for instance particular loci of a human genome. To identify such targets, for example, complexes that each include a catalytically-deactivated DNA binding protein, a guide RNA that guides the complex, and one or more effector domains may be introduced into stem cells to cause at least one of the stem cells to differentiate into a target phenotype. The guide RNAs present in cells demonstrating the target phenotype may be identified and a nucleic acid sequence of each identified guide RNA may be correlated to loci of a genome to identify targets involved in directing cell differentiation to the target phenotype. These methods may be used for directed cell fate specification in stem cells, such as induced pluripotent stem cells, to produce synthetic cells with a desired target phenotype.