种间胚泡互补（IBC）为研究发育提供了独特的平台，并具有克服全球器官短缺的潜力。尽管最近取得了成功，脑组织尚未通过IBC实现。这里,我们开发了一种基于C-CRISPR的优化IBC策略，这有助于快速筛选候选基因，并确定Hesx1缺陷支持通过IBC在小鼠中产生大鼠前脑组织。成年小鼠的异种大鼠前脑组织结构和功能完整。跨物种比较分析显示，大鼠前脑组织的发育速度与小鼠宿主相同，但保持了类似大鼠的转录组特征。随着发育的进行，大鼠细胞的嵌合率逐渐降低，提示产前发育中晚期的异种障碍。种间前脑互补为研究大脑发育和认知功能的进化保守和不同机制打开了大门。基于C-CRISPR的IBC策略在拓宽种间器官形成的研究和应用方面具有巨大潜力。
Interspecies blastocyst complementation (IBC) provides a unique platform to study development and holds the potential to overcome worldwide organ shortages. Despite recent successes, brain tissue has not been achieved through IBC. Here, we developed an optimized IBC strategy based on C-CRISPR, which facilitated rapid screening of candidate genes and identified that Hesx1 deficiency supported the generation of rat forebrain tissue in mice via IBC. Xenogeneic rat forebrain tissues in adult mice were structurally and functionally intact. Cross-species comparative analyses revealed that rat forebrain tissues developed at the same pace as the mouse host but maintained rat-like transcriptome profiles. The chimeric rate of rat cells gradually decreased as development progressed, suggesting xenogeneic barriers during mid-to-late pre-natal development. Interspecies forebrain complementation opens the door for studying evolutionarily conserved and divergent mechanisms underlying brain development and cognitive function. The C-CRISPR-based IBC strategy holds great potential to broaden the study and application of interspecies organogenesis.
