MIC-Drop: A platform for large-scale in vivo CRISPR screens
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MIC-Drop: A platform for large-scale in vivo CRISPR screens. / Parvez, Saba; Herdman, Chelsea; Beerens, Manu; Chakraborti, Korak; Harmer, Zachary P; Yeh, Jing-Ruey J; MacRae, Calum A; Yost, H Joseph; Peterson, Randall T.
In: SCIENCE, Vol. 373, No. 6559, 03.09.2021, p. 1146-1151.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - MIC-Drop: A platform for large-scale in vivo CRISPR screens
AU - Parvez, Saba
AU - Herdman, Chelsea
AU - Beerens, Manu
AU - Chakraborti, Korak
AU - Harmer, Zachary P
AU - Yeh, Jing-Ruey J
AU - MacRae, Calum A
AU - Yost, H Joseph
AU - Peterson, Randall T
PY - 2021/9/3
Y1 - 2021/9/3
N2 - CRISPR-Cas9 can be scaled up for large-scale screens in cultured cells, but CRISPR screens in animals have been challenging because generating, validating, and keeping track of large numbers of mutant animals is prohibitive. Here, we introduce Multiplexed Intermixed CRISPR Droplets (MIC-Drop), a platform combining droplet microfluidics, single-needle en masse CRISPR ribonucleoprotein injections, and DNA barcoding to enable large-scale functional genetic screens in zebrafish. The platform can efficiently identify genes responsible for morphological or behavioral phenotypes. In one application, we showed that MIC-Drop could identify small-molecule targets. Furthermore, in a MIC-Drop screen of 188 poorly characterized genes, we discovered several genes important for cardiac development and function. With the potential to scale to thousands of genes, MIC-Drop enables genome-scale reverse genetic screens in model organisms.
AB - CRISPR-Cas9 can be scaled up for large-scale screens in cultured cells, but CRISPR screens in animals have been challenging because generating, validating, and keeping track of large numbers of mutant animals is prohibitive. Here, we introduce Multiplexed Intermixed CRISPR Droplets (MIC-Drop), a platform combining droplet microfluidics, single-needle en masse CRISPR ribonucleoprotein injections, and DNA barcoding to enable large-scale functional genetic screens in zebrafish. The platform can efficiently identify genes responsible for morphological or behavioral phenotypes. In one application, we showed that MIC-Drop could identify small-molecule targets. Furthermore, in a MIC-Drop screen of 188 poorly characterized genes, we discovered several genes important for cardiac development and function. With the potential to scale to thousands of genes, MIC-Drop enables genome-scale reverse genetic screens in model organisms.
KW - Animals
KW - CRISPR-Cas Systems
KW - Cardiovascular System/growth & development
KW - Cell Culture Techniques
KW - Genetic Testing
KW - High-Throughput Nucleotide Sequencing
KW - Microfluidic Analytical Techniques
KW - Zebrafish/genetics
U2 - 10.1126/science.abi8870
DO - 10.1126/science.abi8870
M3 - SCORING: Journal article
C2 - 34413171
VL - 373
SP - 1146
EP - 1151
JO - SCIENCE
JF - SCIENCE
SN - 0036-8075
IS - 6559
ER -