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Cas-9 expressing stocks available in the Fly Facility

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 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.

 

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.

 

Nickase Cas-9

 

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 Cas9 construct on the X is marked with RFP. Loss of lig4 activity promotes homologous recombination (HR) over non-homologous end joining (NHEJ). 

 

  • lig4;;nos-Gal4 UAS-cas9 (Stock KK0012)

Genotype:      lig4[169];; nanosGal4VP16 UAS-cas9

Ubiquitously expresses Cas9 under control of nanos regulatory sequences in a lig4 loss of function background. The Cas9 construct is on the 3rd chromosome 

Contact Information

Tiny fly

E-mail contacts

Simon Collier (Fly Facility Manager): s.collier@gen.cam.ac.uk

General Fly Facility enquiries: flyadmin@gen.cam.ac.uk

Microinjection enquiries: flyinject@gen.cam.ac.uk

Fly food enquiries: flymedia@gen.cam.ac.uk

Fly stock requests: gh19@gen.cam.ac.uk

Address

Department of Genetics,
University of Cambridge, Downing Street,
Cambridge CB2 3EH,
United Kingdom

Phone: +44 (0)1223 765124
Fax: +44 (0)1223 333992

 

The Fly Facility webpages are maintained by Simon Collier. Please contact me ) if you have any problems using the site.