Lalaouna David, Prévost Karine, Eyraud Alex, Massé Eric
Department of Biochemistry, RNA Group, Université de Sherbrooke, Sherbrooke, Québec J1E 4K8, Canada.
Department of Biochemistry, RNA Group, Université de Sherbrooke, Sherbrooke, Québec J1E 4K8, Canada.
Methods. 2017 Mar 15;117:28-34. doi: 10.1016/j.ymeth.2016.11.011. Epub 2016 Nov 19.
Recent advances in high-throughput sequencing have led to an explosion in the rate of small regulatory RNAs (sRNAs) discovery among bacteria. However, only a handful of them are functionally characterized. Most of the time, little to no targets are known. In Lalaouna et al. (2015), we proposed a new technology to uncover sRNAs targetome, which is based on the MS2-affinity purification (MAPS). We were able to prove its efficiency by applying it on well-characterized sRNAs of Escherichia coli. Thereafter, we adapted the procedure to other kind of RNA (mRNAs and tRNA-derived RNA fragments) and bacteria (pathogenic or Gram-positive strains). Here, we clearly report all improvements and adjustments made to MAPS technology since it was originally reported.
高通量测序技术的最新进展使得细菌中小调节RNA(sRNA)的发现率呈爆发式增长。然而,其中只有少数具有功能特征。大多数情况下,几乎没有已知的靶标。在拉拉乌纳等人(2015年)的研究中,我们提出了一种基于MS2亲和纯化(MAPS)的新技术来揭示sRNA的靶标组。我们通过将其应用于大肠杆菌中已充分表征的sRNA来证明其有效性。此后,我们将该方法应用于其他类型的RNA(mRNA和tRNA衍生的RNA片段)以及细菌(致病菌株或革兰氏阳性菌株)。在此,我们清晰地报告了自最初报道以来对MAPS技术所做的所有改进和调整。