The cell-type-specific recording and manipulation is instrumental to disentangle causal neural mechanisms in physiology and behavior and increasingly requires intersectional control; however, current approaches are largely limited by the number of intersectional features, incompatibility of common effectors and insufficient gene expression. Here, we utilized the protein-splicing technique mediated by intervening sequences (intein) and devised an intein-based intersectional synthesis of transactivator (IBIST) to selectively control gene expression of common effectors in multiple-feature defined cell types in mice. We validated the specificity and sufficiency of IBIST to control fluorophores, optogenetic opsins and Ca²⁺ indicators in various intersectional conditions. The IBIST-based Ca²⁺ imaging showed that the IBIST can intersect five features and that hippocampal neurons tune differently to distinct emotional stimuli depending on the pattern of projection targets. Collectively, the IBIST multiplexes the capability to intersect cell-type features and controls common effectors to effectively regulate gene expression, monitor and manipulate neural activities.

特定于细胞类型的记录和操作有助于解开生理和行为中的因果神经机制，并且越来越需要交叉控制；然而，目前的方法在很大程度上受到交叉特征数量、常见效应器不兼容和基因表达不足的限制。在这里，我们利用由干预序列（内含肽）介导的蛋白质剪接技术，并设计了一种基于内含肽的反式激活因子交叉合成（IBIST），以选择性地控制小鼠多特征定义细胞类型中常见效应子的基因表达。我们验证了 IBIST在各种交叉条件下控制荧光团、光遗传学视蛋白和 Ca ^{2+指标的特异性和充分性。}基于 IBIST 的 Ca ²⁺成像显示，IBIST 可以与五个特征相交，并且海马神经元根据投射目标的模式对不同的情绪刺激进行不同的调整。总的来说，IBIST 多路复用了交叉细胞类型特征和控制常见效应器的能力，以有效调节基因表达、监测和操纵神经活动。