High Efficiency In Vivo Genome Engineering with a Simplified 15-RVD GoldyTALEN DesignReportar como inadecuado




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Transcription activator-like effector nucleases TALENs enable genome engineering in cell culture and many organisms. Recently, the GoldyTALEN scaffold was shown to readily introduce mutations in zebrafish Danio rerio and livestock through non-homologous end joining NHEJ and homology-directed repair HDR. To deploy the GoldyTALEN system for high-throughput mutagenesis in model organisms, a simple design with high efficacy is desirable. We tested the in vivo efficacy of a simplified 15-RVD GoldyTALEN design spacer between 13–20 bp and T nucleotide preceding each TALEN binding site in zebrafish. All 14 tested TALEN pairs 100% introduced small insertions and deletions at somatic efficacy ranging from 24 to 86%, and mutations were inheritable at high frequencies 18–100%. By co-injecting two GoldyTALEN pairs, inheritable deletions of a large genomic fragment up to 18 kb were successfully introduced at two different loci. In conclusion, these high efficiency 15-RVD GoldyTALENs are useful for high-throughput mutagenesis in diverse application including hypothesis testing from basic science to precision medicine.



Autor: Alvin C. Ma, Han B. Lee, Karl J. Clark, Stephen C. Ekker

Fuente: http://plos.srce.hr/



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