PUBLICATION

MIC-Drop: A platform for large-scale in vivo CRISPR screens

Authors
Parvez, S., Herdman, C., Beerens, M., Chakraborti, K., Harmer, Z.P., Yeh, J.J., MacRae, C.A., Yost, H.J., Peterson, R.T.
ID
ZDB-PUB-210821-3
Date
2021
Source
Science (New York, N.Y.)   373(6559): 1146-1151 (Journal)
Registered Authors
MacRae, Calum A., Peterson, Randall, Yost, H. Joseph
Keywords
none
MeSH Terms
  • Genetic Testing*
  • High-Throughput Nucleotide Sequencing
  • Animals
  • Cardiovascular System/growth & development
  • Zebrafish/genetics*
  • Zebrafish/growth & development
  • Microfluidic Analytical Techniques*
  • Cell Culture Techniques
  • CRISPR-Cas Systems*
(all 9)
PubMed
34413171 Full text @ Science
Abstract
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 report 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 show MIC-Drop can identify small molecule targets. Furthermore, in a MIC-Drop screen of 188 poorly characterized genes, we discover 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.
Genes / Markers
Figures
Figure Gallery (4 images)
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Expression
No data available
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
f2TgTransgenic Insertion
    twu34TgTransgenic Insertion
      1 - 2 of 2
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      Target Reagent Reagent Type
      actb1CRISPR1-actb1CRISPR
      actb1CRISPR2-actb1CRISPR
      actb2CRISPR1-actb2CRISPR
      actb2CRISPR2-actb2CRISPR
      aladCRISPR1-aladCRISPR
      aladCRISPR2-aladCRISPR
      aladCRISPR3-aladCRISPR
      aladCRISPR4-aladCRISPR
      atp6v1c1aCRISPR1-atp6v1c1aCRISPR
      atp6v1c1aCRISPR2-atp6v1c1aCRISPR
      1 - 10 of 78
      Show
      Fish
      No data available
      Antibodies
      No data available
      Orthology
      No data available
      Engineered Foreign Genes
      Marker Marker Type Name
      DsRed2EFGDsRed2
      EGFPEFGEGFP
      1 - 2 of 2
      Show
      Mapping
      No data available
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      Citation
      Parvez, S., Herdman, C., Beerens, M., Chakraborti, K., Harmer, Z.P., Yeh, J.J., MacRae, C.A., Yost, H.J., Peterson, R.T. (2021) MIC-Drop: A platform for large-scale in vivo CRISPR screens. Science (New York, N.Y.). 373(6559):1146-1151.