TY - JOUR
T1 - CRISPR-based genetic diagnostics in microgravity
AU - Alon, Dan Mark
AU - Mittelman, Karin
AU - Stibbe, Eytan
AU - Countryman, Stefanie
AU - Stodieck, Louis
AU - Doraisingam, Shankini
AU - Leal Martin, Dylan Mikeala
AU - Hamo, Eliran Raphael
AU - Pines, Gur
AU - Burstein, David
N1 - Publisher Copyright: © 2023 The Authors
PY - 2023/10/1
Y1 - 2023/10/1
N2 - Monitoring astronauts' health during space missions poses many challenges, including rapid assessment of crew health conditions. Sensitive genetic diagnostics are crucial for examining crew members and the spacecraft environment. CRISPR-Cas12a, coupled with isothermal amplification, has proven to be a promising biosensing system for rapid, on-site detection of genomic targets. However, the efficiency and sensitivity of CRISPR-based diagnostics have never been tested in microgravity. We tested the use of recombinase polymerase amplification (RPA) coupled with the collateral cleavage activity of Cas12a for genetic diagnostics onboard the International Space Station. We explored the detection sensitivity of amplified and unamplified target DNA. By coupling RPA with Cas12a, we identified targets in attomolar concentrations. We further assessed the reactions' stability following long-term storage. Our results demonstrate that CRISPR-based detection is a powerful tool for on-site genetic diagnostics in microgravity, and can be further utilized for long-term space endeavors to improve astronauts' health and well-being.
AB - Monitoring astronauts' health during space missions poses many challenges, including rapid assessment of crew health conditions. Sensitive genetic diagnostics are crucial for examining crew members and the spacecraft environment. CRISPR-Cas12a, coupled with isothermal amplification, has proven to be a promising biosensing system for rapid, on-site detection of genomic targets. However, the efficiency and sensitivity of CRISPR-based diagnostics have never been tested in microgravity. We tested the use of recombinase polymerase amplification (RPA) coupled with the collateral cleavage activity of Cas12a for genetic diagnostics onboard the International Space Station. We explored the detection sensitivity of amplified and unamplified target DNA. By coupling RPA with Cas12a, we identified targets in attomolar concentrations. We further assessed the reactions' stability following long-term storage. Our results demonstrate that CRISPR-based detection is a powerful tool for on-site genetic diagnostics in microgravity, and can be further utilized for long-term space endeavors to improve astronauts' health and well-being.
KW - Amplification
KW - Cas12
KW - DNA
KW - Detection
KW - Space
UR - http://www.scopus.com/inward/record.url?scp=85165026030&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.bios.2023.115479
DO - https://doi.org/10.1016/j.bios.2023.115479
M3 - مقالة
C2 - 37459685
SN - 0956-5663
VL - 237
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
M1 - 115479
ER -