通过与扫描阵列杂交筛选出的反义寡核苷酸是有效的体内试剂。
Antisense oligonucleotides selected by hybridisation to scanning arrays are effective reagents in vivo.
作者信息
Sohail M, Hochegger H, Klotzbücher A, Guellec R L, Hunt T, Southern E M
机构信息
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
出版信息
Nucleic Acids Res. 2001 May 15;29(10):2041-51. doi: 10.1093/nar/29.10.2041.
Abstract
Transcripts representing mRNAs of three Xenopus cyclins, B1, B4 and B5, were hybridised to arrays of oligonucleotides scanning the first 120 nt of the coding region to assess the ability of the immobilised oligonucleotides to form heteroduplexes with their targets. Oligonucleotides that produced high heteroduplex yield and others that showed little annealing were assayed for their effect on translation of endogenous cyclin mRNAs in Xenopus egg extracts and their ability to promote cleavage of cyclin mRNAs in oocytes by RNase H. Excellent correlation was found between antisense potency and affinity of oligonucleotides for the cyclin transcripts as measured by the array, despite the complexity of the cellular environment.
摘要
代表三种非洲爪蟾细胞周期蛋白（B1、B4和B5）mRNA的转录本与扫描编码区前120个核苷酸的寡核苷酸阵列进行杂交，以评估固定化寡核苷酸与其靶标形成异源双链体的能力。检测产生高异源双链体产量的寡核苷酸以及退火很少的其他寡核苷酸对非洲爪蟾卵提取物中内源性细胞周期蛋白mRNA翻译的影响，以及它们促进卵母细胞中细胞周期蛋白mRNA被核糖核酸酶H切割的能力。尽管细胞环境复杂，但通过阵列测量发现，寡核苷酸的反义效力与其对细胞周期蛋白转录本的亲和力之间存在极好的相关性。
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