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Ribosome Profiling

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...

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Updated: Jun 9, 2025

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Published on: November 1, 2019

基于活动的DNA编码图书馆选真核转化选择性抑制剂.

Huda Barhoosh¹, Anjali Dixit¹, Wesley G Cochrane¹

¹Department of Pharmaceutical Sciences, University of California, Irvine, California 92697, United States.

ACS central science

|October 28, 2024

概括

此摘要是机器生成的。

研究人员开发了一种使用体外转录-翻译 (IVTT) 和DNA编码图书馆 (DEL) 的新方法,以发现小分子探针,用于以前无法使用的蛋白质. 这个平台可以在人类蛋白质组中进行可扩展的探头发现.

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MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells

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Last Updated: Jun 9, 2025

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Published on: November 1, 2019

Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling

Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling

Published on: October 7, 2021

MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells

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Published on: December 21, 2011

科学领域:

生物化学生物化学
分子生物学分子生物学
药物发现药物发现药物发现

背景情况:

大多数人体蛋白质缺乏可用药物的结合口袋,将小分子探针的发展限制在蛋白质组的2%左右.
对于许多蛋白质标,传统的高通量选试验往往是不可行的.

研究的目的:

开发一种通用选试验,用于发现针对具有挑战性或未知功能的蛋白质的小分子探针.
为了规避法定药物适用性和测试开发的局限性.

主要方法:

通过将记者基因 (例如GFP) 融合到目标序列,开发了一种体外转录-翻译 (IVTT) 活性试验.
通过使用微流体 picoliter 尺度滴滴,选了 5,348 个成员的 DNA 编码库 (DEL) 寻找翻译抑制剂.
使用PCSK9-GFP报告员验证的命中,包括细胞分析和初步选择性分析.

主要成果:

确定了PCSK9-GFP IVTT的多个抑制剂,其化合物降低了HepG2细胞中的PCSK9水平.
通过选DEL与RPL27,KRASG12D,MST1和USO1.1等多种目标来证明一种插入和使用的方法.
在1-20μM之间实现了IC50值,用于验证的命中.

结论:

微流体IVTT DEL选平台提供了一个可扩展的解决方案,用于对更广泛的蛋白质标进行探针发现.
这种方法显著扩大了识别人类蛋白质组及其以外的化学探针的潜力.
选择性翻译调制为克服传统药物发现约束提供了一个可行的策略.