Fallon Tess K, Knouse Kristin A
Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Cell Genom. 2025 Mar 12;5(3):100777. doi: 10.1016/j.xgen.2025.100777. Epub 2025 Feb 24.
Genome-wide CRISPR screening in the organism has tremendous potential to answer long-standing questions of mammalian physiology and disease. However, bringing this powerful technology in vivo presents unique challenges, including delivering a genome-wide sgRNA library to the appropriate cell type, achieving sufficient coverage of the library, and selecting for the phenotype of interest. In this review, we highlight recent advances in sgRNA delivery, library design, and phenotypic readout that can help overcome these technical challenges and thereby bring high-throughput genetic dissection to an increasing number of tissues and questions. We are excited about the potential for ongoing innovation in these areas to ultimately enable genome-wide CRISPR screening in any cell type of interest in the organism, allowing for unprecedented investigation into diverse questions of mammalian physiology and disease.
在生物体中进行全基因组CRISPR筛选，对于解答哺乳动物生理学和疾病方面长期存在的问题具有巨大潜力。然而，在体内应用这项强大的技术面临着独特的挑战，包括将全基因组sgRNA文库递送至合适的细胞类型、实现文库的充分覆盖以及筛选出感兴趣的表型。在这篇综述中，我们重点介绍了sgRNA递送、文库设计和表型读出方面的最新进展，这些进展有助于克服这些技术挑战，从而将高通量基因剖析应用于越来越多的组织和问题研究中。我们对这些领域正在进行的创新潜力感到兴奋，这些创新最终可能使在生物体中任何感兴趣的细胞类型中进行全基因组CRISPR筛选成为可能，从而能够以前所未有的方式研究哺乳动物生理学和疾病的各种问题。
