The following information accompanied stocks donated to the Bloomington Stock Center by Fillip Port and Simon Bullock, Medical Research Council, Cambridge.
P{UAS-Cas9.P2} consists of a human codon-optimized Cas9 sequence cloned into the pBID-UASC (FBmc0003100) vector. This sequence is the same used previously in the M{Act5C-Cas9.P} and M{nos-Cas9.P} constructs. The advantage of this construct over P{UAS-Cas9.P} is that Cas9 is expressed at a lower level and causes less toxicity. It also shows less leaky expression in the absence of a GAL4 driver.
P{UAS-Cas9.P2}attP40 is an insertion in P{CaryP}attP40 at 25C6,  2L:5108448  (R6).
P{UAS-Cas9.P2}attP2 is an insertion in P{CaryP}attP2 at 68A4,  3L:11070538  (R6).
P{nos-FokI.Cas9} expresses a FokI endonuclease domain fused to catalytically inactive, human codon-optimized Cas9. The Cas9 sequence is the same as that above, but has been inactivated by amino acid substitutions in the two active sites (D10A and H841A). The nos regulatory sequences and 3’ UTR are the same as those present in M{nos-Cas9.P}. DNA cleavage by FokI endonuclease domains is strictly dimerisation dependent, therefore Fok1-dCas9 leads to heritable genome edits only when recruited by two gRNAs in a defined spacing and orientation. This improves the specificity of genome editing over that obtained with wild type Cas9.
P{Act5C-FokI.Cas9} is a similar construct with Act5C regulatory sequences. The Act5C sequences are the same as those present in M{Act5C-Cas9.P}.
P{nos-FokI.Cas9}attP40 and P{Act5C-FokI.Cas9}attP40 are insertions in P{CaryP}attP40 at 25C6,  2L:5108448  (R6).