The Cas-9 expressing stocks currently available in the Fly Facility are listed below. if the stock you want to inject is not on this list, please contact us at flyinject@gen.cam.ac.uk as we may have received it recently (CRISPR is a fast moving field!). If we do not keep the Cas-9 stock you need, please send some healthy flies for us to inject. Our guidelines to sending stocks for microinjection work are in section 4 of our 'Drosophila Microinjection - All you need to know' page.
Please note: We can send you a few sample flies from any of our CRISPR lines prior to injection to allow you to verify that your target sequence is conserved in our stock.
Cas-9 on X chromosome
Genotype: y1 P(act5c-cas9, w+) M(3xP3-RFP.attP)ZH-2A w*
Expresses Cas9 protein ubiquitously under control of Act5C regulatory sequences. The attP landing site is marked with RFP.
Genotype: y1 P(nos-cas9, w+) M(3xP3-RFP.attP)ZH-2A w*
Expresses Cas9 protein in the germline under control of nanos regulatory sequences. The Cas9 insert is labeled with white+. The attP landing site is marked with RFP.
- nos-cas9 (CFD2) mini-white-, DsRed- (Stock MLJ051 from Matt Jones - Sarah Bray lab)
Genotype: y1 P(nos-cas9, w-) M(RFP-.attP)ZH-2A w*
Expresses Cas9 protein in the germline under control of nanos regulatory sequences. The white+ marker labeling the Cas9 insert has been removed (Cre'd out) as has the RFP (DsRed) marker labeling the attP site.
Genotype: y1 P(UAS-cas9, w+) M(3xP3-RFP.attP)ZH-2A w*, nanosGal4VP16
Expresses Cas9 protein under UAS control. Expresses GAL4 in the germline under control of nanos regulatory sequences. The attP landing site is marked with RFP.
- vas-cas9 (Bloomington: 51323)
Genotype: y[1] M{vas-Cas9}ZH-2A w[1118]
Expresses Cas9 protein in the germline under control of vasa regulatory sequences. The Cas9 insertion is marked with GFP and the attP landing site is marked with RFP.
- vas-cas9 RFP- (Bloomington 55821)
Genotype: y1 M{vas-Cas9.RFP-}ZH-2A w1118
Expresses Cas9 protein in the germline under control of vasa regulatory sequences. The Cas9 insertion is marked with GFP. The RFP marking the attP landing site has been removed.
Cas-9 on 2nd chromosome
- TH_attP40
Genotype: y[1] sc[1] v[1]; {y[+t7.7] v[+t1.8]=nanos-Cas9}attp40
Expresses Cas9 protein in the germline under control of nanos regulatory sequences. The Cas9 insert on the 2nd chromosome makes this stock useful when targeting X chromosome sequences.
Cas-9 on 3rd chromosome
- TH_attP2
Genotype: y[1] sc[1] v[1];; {y[+t7.7] v[+t1.8]=nanos-Cas9}attp2
Expresses Cas9 protein in the germline under control of nanos regulatory sequences. The Cas9 insert on the 3rd chromosome makes this stock useful when targeting X chromosome sequences.
- w;; vas-Cas9(VK00027) (Bloomington 51324)
Genotype: w[1118]; PBac{y[+mDint2]=vas-Cas9}VK00027
Expresses Cas9 protein in the germline under control of vasa regulatory sequences. The Cas9 insert is labeled with 3xP3-GFP and yellow+.
Nickase Cas-9
- nickase cas9 (CFD13)
Genotype: P{ry[+t7.2]=hsFLP}1,y[1]w[1118];;P{y[+t7.7]w[+mC]=act-Cas9D10A}attP2
Ubiquitously expresses a variant of Cas9 protein that nicks a single strand of the DNA target. The Cas9 insert on the 3rd chromosome is marked with yellow+ and white+.
Cas-9 in a lig4 mutant background
- act-cas9, lig4 (Bloomington 58492)
Genotype: y[1] M{Act5C-Cas9.P.RFP-}ZH-2A w[1118] Lig4[169]
Ubiquitously expresses Cas9 in a lig4 loss of function background. The miniwhite and 3xP3-RFP marking the Cas9 insertion site have been removed by cre recombinase. Loss of lig4 activity promotes homologous recombination (HR) over non-homologous end joining (NHEJ).
- lig4;;nos-Gal4 UAS-cas9 (Stock KK0010)
Genotype: lig4[169];; nanosGal4VP16 UAS-cas9
Expresses Cas9 protein in the germline under control of nanos regulatory sequences in a lig4 loss of function background. The Cas9 construct is on the 3rd chromosome
