We provide a protocol for neural blastocyst complementation \(NBC), a method we have developed for studying development and function of speci c forebrain regions.We describe all experimental steps from genetically modifying a locus of interest, via CRISPR/Cas9 editing in mouse embryonic stem cells \ (ESCs), to generating chimeric mice with ESC-reconstituted forebrain regions that can be directly analyzed.Our _in vivo_ brain organogenesis approach is e cient, allowing functional and systematic analysis of genes \(or other factors) in as little as three months in the context of a whole organism.This protocol is related to Chang _et al._ \(DOI number, _Nature_, publishing date)
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Alt et al. (2018) studied this question.
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