Wet Lab RNA SmartCycler (Wet Lab RNA SmartCycler) - 10.11.17

Summary | Overview | Operations | Results | Publications | Imagery

ISS Science for Everyone

Science Objectives for Everyone
Wetlab RNA SmartCycler is a research platform for conducting real-time quantitative gene expression analysis aboard the International Space Station (ISS). The system enables spaceflight genomic studies involving a wide variety of biospecimen types in the unique microgravity environment of space.
Science Results for Everyone Gene analysis goes to space. Tests of WetLab-2’s tools for real-time gene expression analysis in space demonstrated that quantitative polymerase chain reaction (qPCR), RNA extraction, and gene expression analysis by Reverse Transcriptase-qPCR work well in microgravity. The system adapted to problems such as accumulating gas interfering with data uniformity. Threshold values for qPCR and PCR were similar to Earth controls. Analyses of gene expression of RNA extractions successfully completed within 3 hours, allowing prompt transmission of results from multiple biological samples. WetLab-2 provides novel capabilities for design, modification, and repetition of molecular biology experiments in space.

The following content was provided by Macarena Parra, Ph.D., and is maintained in a database by the ISS Program Science Office.
Facility Details

OpNom: Wet Lab RNA SmartCycler

Facility Manager(s)
Rudy Aquilina, NASA Ames Research Center, Moffett Field, CA, United States

Facility Representative(s)
Macarena Parra, Ph.D., NASA Ames Research Center, Moffett Field, CA, United States

Developer(s)
NASA Ames Research Center, Moffett Field, CA, United States

Sponsoring Space Agency
National Aeronautics and Space Administration (NASA)

Sponsoring Organization
National Laboratory (NL)

ISS Expedition Duration
March 2016 - April 2017

Expeditions Assigned
47/48,49/50

Previous Missions
This is the first mission for Wetlab RNA SmartCycler.

Availability
Information Pending

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Facility Description

Facility Overview

Currently, gene expression analyses of space flown biospecimens must be conducted postflight—after living cultures or frozen or chemically fixed samples are returned to Earth from space station. Postflight analysis is limited for several reasons. First changes in gene expression can be transient, changing over a timescale of minutes. The delay between sampling a culture in space and analyzing that sample on Earth may affect results. Lastly, follow up experiments designed in response to postflight results must wait for a new flight opportunity to be tested.
 
Wetlab RNA SmartCycler solves these problems by allowing invetigators to obtain real-time genomic data from samples processed and analyzed aboard the space station,
 
Studies that characterize how spaceflight affects the gene expression of cells microbes and tissues are key to helping researchers to better understand how life is affected by or adapts to spaceflight. Gene expression data helps reveal the molecular and cellular mechanisms involved with spaceflight induced conditions such as bone and muscle loss, impaired immunity and increased microbial virulence. Knowledge of these mechanisms can be applied towards developing countermeasures for protecting human health during long-duration space missions and also for treating diseases on Earth.
 
Wetlab RNA SmartCycler enables traditional uses of quantitative PCR, such as measuring gene transcription or rapid detection of gene targets that indicate infectious disease, cell stress, changes in cell cycle, growth and development and/or genetic abnormality. Applications range from fundamental biology investigations to commercial drug discovery efforts. Wetlab RNA SmartCyclercan also be used for real-time  analysis of air, surface, water or clinical samples to monitor environmental conditions and crew health. It can also be used to validate terrestrial analyses of samples returned from the space station by providing quantitative gene expression benchmarking prior to sample return to Earth.
 
Wetlab RNA SmartCycler includes a commercial quantitative PCR instrument (Cepheid SmartCycler), a sample transfer tool for retrieving samples from culturing hardware and a set of fluidic modules to facilitate sample preparation work that is done by the ISS crew members.
 

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Operations

Facility Operations
Components of the Wet Lab RNA SmartCycler system include a commercial quantitative PCR instrument (Cepheid SmartCycler), a sample transfer tool for retrieving samples from culturing hardware; and a set of fluidic modules to facilitate sample preparation work that will be done by crew members working in a weightless environment.

 

Fluidic modules include: the sample processing module that is designed to lyse cells and extract RNA; the reaction assembly module that removes air bubbles from fluids and transfers liquid samples into PCR reaction tubes that are preloaded with stabilized lyophilized reagents and fitted with specialized caps for use in microgravity.

 

Wet Lab RNA SmartCycler performs up to 16 quantitative PCR reactions in parallel, using up to four optical channels to measure fluorescence. The average time to deliver results is less than four hours.

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Decadal Survey Recommendations

Information Pending

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Results/More Information

NASA facility, WetLab-2, is a suite of tools on the International Space Station designed to process biological samples for real-time gene expression analysis. WetLab-2 includes fluidic RNA Sample Preparation Module, fluid transfer devices, lyophilized (freeze-dried) substances and enzymes for Polymerase Chain Reaction (PCR) assays, a centrifuge, and a real-time PCR thermal cycler. The system intakes diverse biological sample (cells, tissue, surface swabs, blood, etc.), uses temperature-stable substances, and allows for the isolation, purification, and reverse transcription of nucleic acids, such as RNA, for molecular biology analysis. Ground-based and on-orbit tests were conducted to validate the use of WetLab-2, these included E. coli primer assays, lyophilized components tests, sample preparation module tests, hardware tests, quantitative PCR (qPCR) efficiency tests of pre-prepared DNA concentrations (low, mid, high), as well as RNA extraction from E. coli and mouse liver tissue tests. Validation testing demonstrated that qPCR, RNA extraction, and gene expression analysis by Reverse Transcriptase-quantitative PCR (RT-qPCR) work well under microgravity conditions. Additionally, when problems such as gases accumulating in PCR tubes interfered with data uniformity, the system adapted to changes and iterations in real-time. Results showed that qPCR Cycle threshold (Ct) values and PCR efficiencies obtained on-orbit from DNA standards were similar to Earth (1g) controls. On-orbit analyses of gene expression of RNA extractions were successfully completed within 3 hours, thereby allowing researchers to receive results from multiple biological samples promptly. The WetLab-2 facility provides novel capabilities for the conduction of molecular biology research on-orbit, permitting the design, modification, and repetitions of experiments in space. WetLab-2 is publicly available to investigators for research on ISS and may be used to analyze other types of samples or for medical diagnostics.

Results Publications

    Parra MP, Jung J, Boone TD, Tran L, Blaber EA, Brown M, Chin M, Chinn T, Cohen JN, Doebler R, Hoang D, Hyde E, Lera MP, Luzod LT, Mallinson M, Marcu O, Mohamedaly Y, Ricco AJ, Rubins K, Sgarlato GD, Talavera RO, Tong P, Uribe E, Williams JN, Wu D, Yousuf R, Richey CS, Schonfeld J, Almeida EA.  Microgravity validation of a novel system for RNA isolation and multiplex quantitative real time PCR analysis of gene expression on the International Space Station. PLOS ONE. 2017 September 6; 12(9): e0183480. DOI: 10.1371/journal.pone.0183480. PMID: 28877184.

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Ground Based Results Publications

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ISS Patents

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Related Publications

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Related Websites

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Imagery

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The Wetlab-2 system includes a commercial PCR instrument (Cepheid SmartCycler) that can perform up to 16 PCR reactions in parallel. Image courtesy of NASA/Dominic Hart.

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The WetLab-2 RNA Sample De-bubbler and Pipette Loading Device. Image courtesy of NASA/Dominic Hart.

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The WetLab-2 Sample Prep Module (SPM). Image courtesy of NASA/Dominic Hart.

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