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Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in Xenopus

Article DOI: 10.3791/56035-v • 05:34 min • February 1st, 2018
February 1st, 2018 •
1
1Department of Developmental and Cell Biology, University of California, Irvine

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Genes essential for survival pose technical hurdles for creating mutant lines. Leapfrogging circumvents lethality by combining genome editing with primordial germ cell transplantation to create wild-type animals carrying germline mutations. Leapfrogging also permits the efficient generation of homozygous null mutants in the F1 generation. Here, the transplantation step is demonstrated.

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Primordial Germ Cell Transplantation CRISPR/Cas9-based Leapfrogging Xenopus Mutant Lines Embryogenesis Developmental Genetics F1 Generation Microinject Cas9-sgRNA Complexes Blastula Stage Nine Transplantation Uninjected Embryo PGC Donor Embryo Vitelline Envelopes Surgery Incisions Blastopore
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