Deciphering the Thermodynamic Landscape of CRISPR/Cas9: Insights into Enhancing Gene Editing Precision and Efficiency
解析CRISPR/Cas9的热力学景观：对提高基因编辑精度和效率的见解
摘要 (Abstract)
1. J Phys Chem B. 2024 Sep 5;128(35):8409-8422. doi: 10.1021/acs.jpcb.4c04044. Epub 2024 Aug 27. Deciphering the Thermodynamic Landscape of CRISPR/Cas9: Insights into Enhancing Gene Editing Precision and Efficiency. Kumar A(1)(2), Daripa P(1), Rasool K(1), Chakraborty D(1)(2), Jain N(1)(2), Maiti S(1)(2). Author information: (1)CSIR-Institute of Genomics & Integrative Biology, New Delhi 110025, India. (2)Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India. The thermodynamic landscape of the CRISPR/Cas9 system plays a crucial role in understanding and optimizing the performance of this revolutionary genome-editing technology. In this research, we utilized isothermal titration calorimetry and microscale thermophoresis techniques to thoroughly investigate the thermodynamic properties governing CRISPR/Cas9 interactions. Our findings revealed that the binding between sgRNA and Cas9 is primarily governed by entropy, which compensates for an unfavorable enthalpy change. Conversely, the interaction between the CRISPR RNP complex and the target DNA is characterized by a favorable enthalpy change, offsetting an unfavorable entropy change. Notably, both interactions displayed negative heat capacity changes, indicative of potential hydration, ionization, or structural rearrangements. However, we noted that the involvement of water molecules and counterions in the interactions is minimal, suggesting that structural rearrangements play a significant role in influencing the binding thermodynamics. These results offer a nuanced understanding of the energetic contributions and structural dynamics underlying CRISPR-mediated gene editing. Such insights are invaluable for optimizing the efficiency and specificity of CRISPR-based genome editing applications, ultimately advancing our ability to precisely manipulate genetic material in various organisms for research, therapeutic, and biotechnological purposes. DOI: 10.1021/acs.jpcb.4c04044 PMID: 39190773 [Indexed for MEDLINE]
实验设计与方法 (Experimental Design & Methods)
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实验结果 (Experimental Results)
总结了主要研究进展、关键发现和发展趋势，识别了研究空白和未来研究方向。
数据汇总 (Data Summary)
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结论 (Conclusions)
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实践意义 (Practical Significance)
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