Welcome to CRISPR fly design!
We posted a new preprint: gRNA multiplexing | tissue-specific CRISPR | Cpf1 genome editing. Read it here.
CRISPR/Cas is an exciting new technology for genome engineering. We have generated a set of tools that allows targeted mutagenesis in Drosophila melanogaster with high efficiency. The system consists of transgenic cas9 fly lines and gRNA expression vectors. Plasmids encoding target specific gRNAs can be injected into cas9 embryos or integrated into the genome and crossed to cas9 strains. By choosing appropriate tools it is possible to generate loss-of-function mutations in essential or non-essential genes in a germline restricted or ubiquitous fashion. It is also possible to introduce designer mutations by homology directed repair, ranging from single nucleotide changes to GFP knock-ins. The papers describing our CRISPR/Cas tools is available here and here.
At CRISPRflydesign we believe in collaboration, sharing and open access. We started sending our transgenic cas9 lines to interested labs in July 2013, shortly after generating them. The website was launched in August 2013 to facilitate distribution of materials and to share experiences with their use. It allowed us to communicate some of our key results many months before formal publication. We eventually wrote up a manuscript about our CRISPR system in March 2014 and posted it on the pre-print server Biorxiv at the same time as submitting it to PNAS. A revised version of the manuscript has now been published.
Port F, Chen HM, Lee T, Bullock SL. Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila. PNAS, 2014 Jul 22. link
Port F, Muschalik N, Bullock SL. Systematic Evaluation of Drosophila CRISPR Tools Reveals Safe and Robust Alternatives to Autonomous Gene Drives in Basic Research. G3, 2015 May 20. link
Akbari OS, Bellen HJ, Bier E, Bullock SL, Burt A, Church GM, Cook KR, Duchek P, Edwards OR, Esvelt KM, Gantz VM, Golic KG, Gratz SJ, Harrison MM, Hayes KR, James AA, Kaufman TC, Knoblich J, Malik HS, Matthews KA, O’Connor-Giles KM, Parks AL, Perrimon N, Port F, Russell S, Ueda R, Wildonger J. Biosafety. Safeguarding gene drive experiments in the laboratory. Science, 2015 Aug 28. link