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Brogan, D., Akbari, O. (2019.10.30). CasRx and dCasRx constructs and insertions. 
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The following information accompanied stocks donated to the Bloomington Stock Center by Dan Brogan and Omar Akbari, University of California at San Diego.
P{Ubi-CasRx.A} expresses the CasRx programmable ribonuclease described in Konermann et al., Cell 2018 173(3):665-676.e14, PMID:29551272). Expression is driven ubiquitously under the control of Ubi-p63E 5' regulatory sequences and 3' UTR sequences from the Autographa californica nucleopolyhedrovirus p10 gene. The construct is marked with RFPDsRed.OpIE2.
P{UAS-CasRx.A} expresses CasRx under the control of UASt regulatory sequences with the substitution of 3' UTR sequences from the Autographa californica nucleopolyhedrovirus p10 gene. The construct is marked with RFPDsRed.OpIE2.
P{Ubi-dCasRx.A} expresses the catalytically inactive dCasRx protein described in Konermann et al., Cell 2018 173(3):665-676.e14, PMID:29551272). Both of the HEPN domains carry mutations, these are R239A, H244A for HEPN-1 and R858A, H863A for HEPN-2. Expression is driven ubiquitously under the control of Ubi-p63E 5' regulatory sequences and 3' UTR sequences from the Autographa californica nucleopolyhedrovirus p10 gene. The construct is marked with RFPDsRed.OpIE2.
P{UAS-dCasRx.A} expresses the same catalytically inactive dCasRx protein under the control of UASt regulatory sequences with the substitution of 3' UTR sequences from the Autographa californica nucleopolyhedrovirus p10 gene. The construct is marked with RFPDsRed.OpIE2.
P{Ubi-CasRx.A}attP1, P{UAS-CasRx.A}attP1, P{Ubi-dCasRx.A}attP1 and P{UAS-dCasRx.A}attP1 are insertions into the P{CaryP}attP1 target site at 55C4,  2R:18 ,357,321 (R6).
P{U6-N.RxgRNA} expresses a CasRx-specific guide RNA directed against N ubiquitously under the control of  snRNA:U6:96Ac  regulatory sequences. The construct also contains a mini-white marker.
P{U6-y.RxgRNA} expresses a CasRx-specific guide RNA directed against y ubiquitously under the control of  snRNA:U6:96Ac  regulatory sequences. The construct also contains a mini-white marker.
P{U6-pLUC.RxgRNA} expresses a CasRx-specific guide RNA directed against firefly luciferase, expressed ubiquitously under the control of  snRNA:U6:96Ac  regulatory sequences. The construct also contains a mini-white marker.
P{U6-w.RxgRNA} expresses a CasRx-specific guide RNA directed against w ubiquitously under the control of  snRNA:U6:96Ac  regulatory sequences. The construct also contains a mini-white marker.
P{U6-cn.RxgRNA} expresses a CasRx-specific guide RNA directed against cn ubiquitously under the control of  snRNA:U6:96Ac  regulatory sequences. The construct also contains a mini-white marker.
P{U6-N.RxgRNA}attP2, P{U6-y.RxgRNA}attP2, P{U6-pLUC.RxgRNA}attP2, P{U6-w.RxgRNA}attP2 and P{U6-cn.RxgRNA}attP2 are insertions into the P{CaryP}attP2 target site at 68A4,  3L:11 ,070,538 (R6).
PBac{Ubi-pLUC.T2A.eGFP,Ubi-rLUC} expresses firefly luciferase (pLUC), eGFP and Renilla luciferase (rLUC) under the control of Ubi-p63E regulatory sequences. pLUC and eGFP are encoded by a single transcript with a T2A translation termination sequence and a Autographa californica nucleopolyhedrovirus p10 gene 3' UTR. rLUC is encoded by a transcript with SV40 3' UTR sequences. The construct also contains a mini-white marker.
PBac{Ubi-pLUC.T2A.eGFP,Ubi-rLUC}VK00027 is an insertion into the PBac{y+-attP-9A}VK00027 target site at 89E11,  3R:17 ,052,863 (R6).
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