biostudies-literatureParvez SAmerican Heart AssociationNHLBI NIH HHSNINDS NIH HHSNational Institutes of HealthNIGMS NIH HHSNIH HHS1146-1151https://www.ebi.ac.uk/biostudies/studies/S-EPMC9083377biostudies-literatureUnknown373(6559)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 fuScience (New York, N.Y.)MIC-Drop: A platform for large-scale in vivo CRISPR screens.PMC9083377UM1 HL098160U54 NS112107OD R24OD017870U01 HL098160827636NINDS U54-NS079201R01 GM134069R24 OD0178705R01GM134069-02Herdman CPeterson RTChakraborti KMacRae CAYost HJYeh JJBeerens MParvez SHarmer ZPfalseMIC-Drop: A platform for large-scale in vivo CRISPR screens.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 fu2021-01-01T00:00:00Z2021 Sep2025-04-04T20:22:01.209Z2025-04-04T20:22:01.209ZS-EPMC90833773441317110.1126/science.abi8870