Category: pU6-3

Differential activity of U6 gRNA plasmids

Fillip Port1, Hui-Min Chen2, Tzumin Lee2 and Simon Bullock1 1: MRC-Laboratory of Molecular Biology, Cambridge, UK 2: Janelia Farm Research Campus, HHMI, Ashburn, USA   Delivery of gRNAs for fly CRISPR/Cas genome engineering is most commonly done by either DNA or RNA injection or by creating transgenic gRNA lines.  Plasmids for both DNA injection and gRNA transgenes use polymerase III…

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Highly efficient targeting of yellow with transgenic act-cas9 and pU6-3gRNA

Hui-Min Chen and Tzumin Lee Janelia Farm Research Campus, HHMI Here we report a preliminary test experiment combining the act-cas9 (stock CFD-1) with two different gRNAs targeting yellow expressed from either the U6-2 or U6-3 promoter. The pU6-gRNAs were stably integrated into the genome and transgenic flies were crossed to CFD-1 flies. Offspring inherit a wildtype yellow allele from the…

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First test of stock CFD-5: UAS-cas9

Hui-Min Chen and Tzumin Lee Janelia Farm Research Campus, HHMI   We expressed UAS-cas9 (stock CDF-5) under the control of a germline Gal4 line and crossed these flies to transgenic flies expressing gRNAs targeting yellow under the control of the U6-3 promoter (pU6-3-gRNA y). The gRNA target site was: GGAGCGATATAGTTGGAGCCAGCTGG. Female offspring of this cross was then crossed to yellow…

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