Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Protein and DNA Yield Current Enhancements, Slow Translocations, and an Enhanced Signal-to-Noise Ratio under a Salt Imbalance
ACS Sensors ( IF 8.2 ) Pub Date : 2022-06-15 , DOI: 10.1021/acssensors.2c00479 Lauren S Lastra 1 , Y M Nuwan D Y Bandara 1 , Vinay Sharma 1, 2 , Kevin J Freedman 1
ACS Sensors ( IF 8.2 ) Pub Date : 2022-06-15 , DOI: 10.1021/acssensors.2c00479 Lauren S Lastra 1 , Y M Nuwan D Y Bandara 1 , Vinay Sharma 1, 2 , Kevin J Freedman 1
Affiliation
|
Nanopores are a promising single-molecule sensing device class that captures molecular-level information through resistive or conductive pulse sensing (RPS and CPS). The latter has not been routinely utilized in the nanopore field despite the benefits it could provide, specifically in detecting subpopulations of a molecule. A systematic study was conducted here to study the CPS-based molecular discrimination and its voltage-dependent characteristics. CPS was observed when the cation movement along both electrical and chemical gradients was favored, which led to an ∼3× improvement in SNR (i.e., signal-to-noise ratio) and an ∼8× increase in translocation time. Interestingly, a reversal of the salt gradient reinstates the more conventional resistive pulses and may help elucidate RPS–CPS transitions. The asymmetric salt conditions greatly enhanced the discrimination of DNA configurations including linear, partially folded, and completely folded DNA states, which could help detect subpopulations in other molecular systems. These findings were then utilized for the detection of a Cas9 mutant, Cas9d10a─a protein with broad utilities in genetic engineering and immunology─bound to DNA target strands and the unbound Cas9d10a + sgRNA complexes, also showing significantly longer event durations (>1 ms) than typically observed for proteins.
中文翻译：
蛋白质和 DNA 在盐分不平衡下产生电流增强、缓慢易位和增强的信噪比
纳米孔是一种很有前途的单分子传感设备类别，它通过电阻或导电脉冲传感（RPS 和 CPS）捕获分子水平的信息。尽管后者可以提供好处，特别是在检测分子的亚群方面，但后者并未常规用于纳米孔领域。在这里进行了一项系统的研究，以研究基于 CPS 的分子识别及其电压依赖性特征。当有利于沿电和化学梯度的阳离子运动时，观察到 CPS，这导致 SNR（即信噪比）提高 3 倍，易位时间增加 8 倍。有趣的是，盐梯度的逆转恢复了更传统的电阻脉冲，并可能有助于阐明 RPS-CPS 转换。不对称盐条件极大地增强了对包括线性、部分折叠和完全折叠 DNA 状态在内的 DNA 构型的区分，这有助于检测其他分子系统中的亚群。然后将这些发现用于检测 Cas9 突变体 Cas9d10a（一种在基因工程和免疫学中具有广泛用途的蛋白质）与 DNA 靶链和未结合的 Cas9d10a + sgRNA 复合物结合，同时显示事件持续时间显着延长（>1 ms）比通常观察到的蛋白质。
更新日期：2022-06-15
中文翻译：
蛋白质和 DNA 在盐分不平衡下产生电流增强、缓慢易位和增强的信噪比
纳米孔是一种很有前途的单分子传感设备类别，它通过电阻或导电脉冲传感（RPS 和 CPS）捕获分子水平的信息。尽管后者可以提供好处，特别是在检测分子的亚群方面，但后者并未常规用于纳米孔领域。在这里进行了一项系统的研究，以研究基于 CPS 的分子识别及其电压依赖性特征。当有利于沿电和化学梯度的阳离子运动时，观察到 CPS，这导致 SNR（即信噪比）提高 3 倍，易位时间增加 8 倍。有趣的是，盐梯度的逆转恢复了更传统的电阻脉冲，并可能有助于阐明 RPS-CPS 转换。不对称盐条件极大地增强了对包括线性、部分折叠和完全折叠 DNA 状态在内的 DNA 构型的区分，这有助于检测其他分子系统中的亚群。然后将这些发现用于检测 Cas9 突变体 Cas9d10a（一种在基因工程和免疫学中具有广泛用途的蛋白质）与 DNA 靶链和未结合的 Cas9d10a + sgRNA 复合物结合，同时显示事件持续时间显着延长（>1 ms）比通常观察到的蛋白质。
京公网安备 11010802027423号