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A loxP site was inserted upstream of exon 3. A neomycin resistance cassette with a 5' loxP site was inserted downstream of exon 5. Cre-mediated recombination removed exons 2 through 5.
Two loxP sites were inserted into introns 2 and 5, and a neomycin resistance cassette flanked by two FRT sites was inserted into intron 5 as a positive-selection marker. The diphtheria toxin A (DTA) cassette was used as a negative-selection marker to assist the rate of positive recombinant events. The linearized targeting DNA was electroporated into R1 mouse ES cells (derived from (129X1/SvJ x 129S1/Sv)F1-Kitl+). After G418 selection, genotyping was done via restriction length analysis direct sequencing. Two correct recombinant ES cell clones were microinjected into C57BL/6 blastocysts and transferred to pseudopregnant foster mice. Male chimeras were mated with female Black Swiss mice, and progenies with germline transmission (neo/+) were confirmed by PCR. Heterozygous (Epac1+/−) mice were obtained by crossing with protamine-Cre carrier mice (no further detail available). Wild-type (+/+) and Epac1 null (−/−) mice were generated by breeding heterozygous mice. This was followed by backcrossing to C57BL/6 for N20+.
Yan J, Mei FC, Cheng H, Lao DH, Hu Y, Wei J, Patrikeev I, Hao D, Stutz SJ, Dineley KT, Motamedi M, Hommel JD, Cunningham KA, Chen J, Cheng X. Enhanced leptin sensitivity, reduced adiposity, and improved glucose homeostasis in mice lacking exchange protein directly activated by cyclic AMP isoform 1. Mol Cell Biol. 2013 Mar;33(5):918-26. doi: 10.1128/MCB.01227-12. Epub 2012 Dec 21. (Medline PMID: 23263987)
Singhmar P, Huo X, Eijkelkamp N, Berciano SR, Baameur F, Mei FC, Zhu Y, Cheng X, Hawke D, Mayor F Jr, Murga C, Heijnen CJ, Kavelaars A. Critical role for Epac1 in inflammatory pain controlled by GRK2-mediated phosphorylation of Epac1. Proc Natl Acad Sci U S A. 2016 Mar 15;113(11):3036-41. doi: 10.1073/pnas.1516036113. Epub 2016 Feb 29. (Medline PMID: 26929333)
Singhmar P, Huo X, Li Y, Dougherty PM, Mei F, Cheng X, Heijnen CJ, Kavelaars A. Orally active Epac inhibitor reverses mechanical allodynia and loss of intraepidermal nerve fibers in a mouse model of chemotherapy-induced peripheral neuropathy. Pain. 2018 May;159(5):884-893. doi: 10.1097/j.pain.0000000000001160. (Medline PMID: 29369966)
Wang H, Robichaux WG, Wang Z, Mei FC, Cai M, Du G, Chen J, Cheng X. Inhibition of Epac1 suppresses mitochondrial fission and reduces neointima formation induced by vascular injury. Sci Rep. 2016 Nov 10;6:36552. doi: 10.1038/srep36552. (Medline PMID: 27830723)
Robichaux WG 3rd, Mei FC, Yang W, Wang H, Sun H, Zhou Z, Milewicz DM, Teng BB, Cheng X. Epac1 (Exchange Protein Directly Activated by cAMP 1) Upregulates LOX-1 (Oxidized Low-Density Lipoprotein Receptor 1) to Promote Foam Cell Formation and Atherosclerosis Development. Arterioscler Thromb Vasc Biol. 2020 Dec;40(12):e322-e335. doi: 10.1161/ATVBAHA.119.314238. Epub 2020 Oct 15. (Medline PMID: 33054390)
Liu H, Mei FC, Yang W, Wang H, Wong E, Cai J, Toth E, Luo P, Li YM, Zhang W, Cheng X. Epac1 inhibition ameliorates pathological angiogenesis through coordinated activation of Notch and suppression of VEGF signaling. Sci Adv. 2020 Jan 1;6(1):eaay3566. doi: 10.1126/sciadv.aay3566. (Medline PMID: 31911948)
Colony Surveillance Program and Current Health Reports
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