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
-
- Animals
- CRISPR-Cas Systems*
- Cardiovascular System/growth & development
- Cell Culture Techniques
- Genetic Testing*
- High-Throughput Nucleotide Sequencing
- Microfluidic Analytical Techniques*
- Zebrafish/genetics*
- Zebrafish/growth & development
- PubMed
- 34413171 Full text @ Science
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.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping