ARQiv-HTS, a versatile whole-organism screening platform enabling in vivo drug discovery at high-throughput rates

Research output: Contribution to journalArticle

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Abstract

The zebrafish has emerged as an important model for whole-organism small-molecule screening. However, most zebrafish-based chemical screens have achieved only mid-throughput rates. Here we describe a versatile whole-organism drug discovery platform that can achieve true high-throughput screening (HTS) capacities. This system combines our automated reporter quantification in vivo (ARQiv) system with customized robotics, and is termed 'ARQiv-HTS'. We detail the process of establishing and implementing ARQiv-HTS: (i) assay design and optimization, (ii) calculation of sample size and hit criteria, (iii) large-scale egg production, (iv) automated compound titration, (v) dispensing of embryos into microtiter plates, and (vi) reporter quantification. We also outline what we see as best practice strategies for leveraging the power of ARQiv-HTS for zebrafish-based drug discovery, and address technical challenges of applying zebrafish to large-scale chemical screens. Finally, we provide a detailed protocol for a recently completed inaugural ARQiv-HTS effort, which involved the identification of compounds that elevate insulin reporter activity. Compounds that increased the number of insulin-producing pancreatic beta cells represent potential new therapeutics for diabetic patients. For this effort, individual screening sessions took 1 week to conclude, and sessions were performed iteratively approximately every other day to increase throughput. At the conclusion of the screen, more than a half million drug-treated larvae had been evaluated. Beyond this initial example, however, the ARQiv-HTS platform is adaptable to almost any reporter-based assay designed to evaluate the effects of chemical compounds in living small-animal models. ARQiv-HTS thus enables large-scale whole-organism drug discovery for a variety of model species and from numerous disease-oriented perspectives.
Original languageEnglish (US)
Pages (from-to)2432-2453
Number of pages22
JournalNature Protocols
Volume11
Issue number12
StatePublished - Dec 1 2016

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Zebrafish
Drug Discovery
Screening
Throughput
Insulin
High-Throughput Screening Assays
Insulin-Secreting Cells
Robotics
Practice Guidelines
Sample Size
Ovum
Larva
Embryonic Structures
Animal Models
Assays
Chemical compounds
Pharmaceutical Preparations
Titration
Animals
Molecules

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ARQiv-HTS, a versatile whole-organism screening platform enabling in vivo drug discovery at high-throughput rates. / Rajpurohit, Surendra Kumar.

In: Nature Protocols, Vol. 11, No. 12, 01.12.2016, p. 2432-2453.

Research output: Contribution to journalArticle

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