A transient CRISPR/Cas9 expression system for genome editing in Trypanosoma brucei.

作者: Sebastian Shaw , Sebastian Knüsel , Sarah Hoenner , Isabel Roditi

DOI: 10.1186/S13104-020-05089-Z

关键词:

摘要: Generation of knockouts and in situ tagging genes Trypanosoma brucei has been greatly facilitated by using CRISPR/Cas9 as a genome editing tool. To date, this entailed limited number cell lines that are stably transformed to express Cas9 T7 RNA polymerase (T7RNAP). It would be desirable, however, able use for any trypanosome line. We describe sequential transfection expression system enables transient the two proteins, followed delivery PCR products gRNAs repair templates. This procedure can used without need stable integration T7RNAP genes.

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