Abstract: Provided herein, in some embodiments, are nucleic acid-based tools that may be used for high-throughput functional genomics studies as well as for the generation of knockout (gene inactivation or deletion) or knockin (gene activation or insertion) cell lines. Tools of the present disclosure include an “activatable reporter cassette,” a guide RNA construct and a nuclease that can be used together, for example, to modify and isolate targeted cells of interest.

Abstract: The present disclosure provides a non-naturally occurring CRISPR-Cas system comprising: a Cas9 effector protein capable of generating cohesive ends (stiCas9), and a guide polynucleotide that forms a complex with the stiCas9 and comprising a guide sequence, wherein the guide sequence hybridizes with a target sequence in a eukaryotic cell but does not hybridize to a sequence in a bacterial cell, and wherein the complex does not occur in nature. The present disclosure also provides a method of introducing a sequence of interest into a chromosome of a cell. Finally, the present disclosure provides for a method of modifying one or more nucleotides using seamless mutagenesis.

Abstract: Provided herein, in some embodiments, are nucleic acid-based tools that may be used for high-throughput functional genomics studies as well as for the generation of knockout (gene inactivation or deletion) or knockin (gene activation or insertion) cell lines. Tools of the present disclosure include an “activatable reporter cassette,” a guide RNA construct and a nuclease that can be used together, for example, to modify and isolate targeted cells of interest.