Deciphering Post-transcriptional Gene Regulatory Networks During Periods of Host-Pathogen Interaction and Innate Immune Activation

项目摘要 我们正在申请父母资助R35GM119569的补充资金，以购买Beckman-Coulter 超速离心机(Optima XE-100 IVD)和两个转子(45Ti和SW32Ti)。阿斯卡诺实验室经常 利用密度梯度超速离心法浓缩病毒并将其从细胞中分离出来 传播过程中的碎片。然而，目前的超离心机(Beckman Optima le-80K)已超过19岁 (于2003年购买)，在过去两年中不得不进行两次维修，在此期间，它经历了 重大停机时间，因为陈旧设备的更换部件更难采购-AS 由维修仪器的Beckman维修技术人员进行沟通。考虑到它的年龄，修复 目前的工具只会继续增加。此外，实验室目前使用的两个转子，一个45Ti和一个 SW32Ti，也是较旧的-最新的(SW32Ti)是在2014年购买的；两个转子都不再是 在保修期内。我们主要是在申请资金，以获得一种新的工具，因为我们预见到不可避免的情况 目前的部门超速离心机退役。超离心机是任务关键的一块 使实验室能够分离和浓缩各种RNA病毒以达到高滴度和高纯度的设备。这 是制备我们最重要的生物试剂之一的必要和必要的一步，我们需要 R35父母资助的四个研究方向中的三个。

Project Summary We are requesting supplemental funds to the parental grant R35GM119569 to purchase a Beckman-Coulter ultracentrifuge (Optima XE-100 IVD) and two rotors (45Ti and SW32Ti). The Ascano laboratory routinely utilizes density gradient ultracentrifugation as a means of concentrating viruses and isolating them from cellular debris during propagation. However, the current ultracentrifuge (Beckman Optima LE-80K) is over 19 years old (was purchased in 2003) and has had to be serviced twice in the last two years, during which it experienced significant downtimes because replacement parts for an obsolete piece of equipment are harder to source – as communicated by Beckman repair technicians servicing the instrument. Given its age, the costs to repair the current instrument only continue to increase. Moreover, the two rotors that the lab currently use, a 45Ti and a SW32Ti, are also older - with the newest one (SW32Ti) being purchased in 2014; both rotors are no longer under warranty. We are principally requesting funds to secure a new instrument, in anticipation of the inevitable decommissioning of the current departmental ultracentrifuge. An ultracentrifuge is a mission-critical piece of equipment enabling the laboratory to isolate and concentrate various RNA viruses to high titer and purity. This is a necessary and essential step for preparing one of our most important biological reagents, which we require for three of four research directions of the R35 parental grant.