申请人:The University of Chicago
公开号:US11136618B2
公开(公告)日:2021-10-05
Described herein are nucleic acid molecules and complexes useful as i-switch pH reporters that have increased sensitivities as a pH reporter and have alternate pH reporting capacity ranges. Aspects of the disclosure relate to a method for determining pH comprising providing a nucleic acid complex comprising: a first single-stranded nucleic acid molecule comprising the sequence CnXCnYCnZCn (SEQ ID NO. 6) wherein C is cytosine; X, Y and Z are each one or more of adenine, thymine, guanine, or combinations thereof; and n is greater than or equal to 2; and wherein at least 2 cytosine residues of the first single-stranded nucleic acid molecule are modified; and a second single-stranded nucleic acid molecule that is partially or fully complementary to the first single-stranded molecule, wherein a first label is conjugated to the first single-stranded nucleic acid molecule or the second single-stranded nucleic acid molecule; and wherein the first label is capable of producing a signal, wherein the intensity of the signal varies as a function of the conformation of the nucleic acid complex; and measuring the intensity of the signal and determining the pH from the measured signal.
本文描述了可用作 i 开关 pH 报告器的核酸分子和复合物,它们作为 pH 报告器具有更高的灵敏度,并具有替代的 pH 报告能力范围。本公开的各个方面涉及一种测定 pH 值的方法,该方法包括提供一种核酸复合物,该核酸复合物包含:第一单链核酸分子,该核酸分子包含序列 CnXCnYCnZCn(SEQ ID NO.6),其中 C 是胞嘧啶;X、Y 和 Z 分别是腺嘌呤、胸腺嘧啶、鸟嘌呤或其组合中的一种或多种;以及 n 大于或等于 2;并且其中第一单链核酸分子的至少 2 个胞嘧啶残基被修饰;和与第一单链分子部分或完全互补的第二单链核酸分子,其中第一标签与第一单链核酸分子或第二单链核酸分子共轭;其中第一标签能够产生信号,信号强度随核酸复合物构象的变化而变化;以及测量信号强度并根据测量的信号确定 pH 值。