CRISPR guide RNAs (gRNAs) can be programmed with relative ease to allow the genetic editing of nearly any DNA or RNA sequence. Here, we propose novel molecular architectures to achieve RNA-dependent modulation of CRISPR activity in response to specific RNA molecules. We designed and tested, in both living Escherichia coli cells and cell-free assays for rapid prototyping, cis-repressed RNA-interacting guide RNA (igRNA) that switch to their active state only upon interaction with small RNA fragments or long RNA transcripts, including pathogen-derived mRNAs of medical relevance such as the human immunodeficiency virus infectivity factor. The proposed CRISPR-igRNAs are fully customizable and easily adaptable to the majority if not all the available CRISPR-Cas variants to modulate a variety of genetic functions in response to specific cellular conditions, providing orthogonal activation and increased specificity. We thereby foresee a large scope of application for therapeutic, diagnostic, and biotech applications in both prokaryotic and eukaryotic systems.

中文翻译：

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用于遗传传感和诊断的工程化 RNA 相互作用 CRISPR 引导 RNA


CRISPR 引导 RNA (gRNA) 可以相对轻松地进行编程，以允许对几乎任何 DNA 或 RNA 序列进行基因编辑。在这里，我们提出了新颖的分子结构，以响应特定 RNA 分子来实现 CRISPR 活性的 RNA 依赖性调节。我们在活体大肠杆菌细胞和用于快速原型制作的无细胞测定中设计并测试了顺式抑制的 RNA 相互作用指导 RNA (igRNA)，仅在与小 RNA 片段或长 RNA 转录物相互作用时才切换到活性状态，包括具有医学相关性的病原体衍生的 mRNA，例如人类免疫缺陷病毒感染因子。所提出的 CRISPR-igRNA 是完全可定制的，并且很容易适应大多数（如果不是全部）可用的 CRISPR-Cas 变体，以响应特定的细胞条件调节各种遗传功能，提供正交激活和增加的特异性。因此，我们预见了原核和真核系统中治疗、诊断和生物技术应用的广泛应用。