Center for Human Genetics, KU Leuven, Leuven, Belgium.
Center for Cancer Biology, VIB, Leuven, Belgium.
J Hematol Oncol. 2023 Oct 24;16(1):107. doi: 10.1186/s13045-023-01495-5.
The CRISPR genome editing technology has revolutionized the way gene function is studied. Genome editing can be achieved in single genes or for thousands of genes simultaneously in sensitive genetic screens. While conventional genetic screens are limited to bulk measurements of cell behavior, recent developments in single-cell technologies make it possible to combine CRISPR screening with single-cell profiling. In this way, cell behavior and gene expression can be monitored simultaneously, with the additional possibility of including data on chromatin accessibility and protein levels. Moreover, the availability of various Cas proteins leading to inactivation, activation, or other effects on gene function further broadens the scope of such screens. The integration of single-cell multi-omics approaches with CRISPR screening open the path to high-content information on the impact of genetic perturbations at single-cell resolution. Current limitations in cell throughput and data density need to be taken into consideration, but new technologies are rapidly evolving and are likely to easily overcome these limitations. In this review, we discuss the use of bulk CRISPR screening in hematology research, as well as the emergence of single-cell CRISPR screening and its added value to the field.
CRISPR 基因组编辑技术彻底改变了研究基因功能的方式。基因组编辑可以在单个基因中实现，也可以在敏感的遗传筛选中同时实现数千个基因。虽然传统的遗传筛选仅限于细胞行为的批量测量，但单细胞技术的最新发展使得将 CRISPR 筛选与单细胞分析相结合成为可能。通过这种方式，可以同时监测细胞行为和基因表达，并且有可能包括有关染色质可及性和蛋白质水平的数据。此外，各种 Cas 蛋白的可用性导致基因功能的失活、激活或其他影响，进一步扩大了此类筛选的范围。单细胞多组学方法与 CRISPR 筛选的整合为在单细胞分辨率下研究遗传扰动对细胞的影响提供了高信息量的途径。需要考虑细胞通量和数据密度方面的当前限制，但新技术正在迅速发展，很可能轻易克服这些限制。在这篇综述中，我们讨论了在血液学研究中使用批量 CRISPR 筛选，以及单细胞 CRISPR 筛选的出现及其对该领域的附加值。
