172: 84760. 39. Hirano T, Funahashi SI, Uemura T, Yanagida M Isolation and characterization of Schizosaccharomyces pombe cut mutants that block nuclear division but not cytokinesis. EMBO J 5: 2973979. 40. Uzawa S, Samejima I, Hirano T, Tanaka K, Yanagida M The fission yeast cut1+ gene regulates spindle pole body duplication and has homology to the budding yeast ESP1 gene. Cell 62: 91325. 41. Stratmann R, Lehner CF Separation of sister chromatids in mitosis requires the Drosophila pimples product, a protein degraded after the metaphase/anaphase transition. Cell 84: 255. 42. Mei J, Huang X, Zhang P Securin is not required for cellular viability, but is required for normal growth of mouse embryonic fibroblasts. Curr Biol 11: 1197201. 43. Wang Z, Yu R, Melmed S Mice lacking pituitary tumor transforming gene show testicular and splenic hypoplasia, thymic hyperplasia, thrombocytopenia, aberrant cell cycle progression, and premature centromere division. Mol Endocrinol 15: 1870879. 44. Jallepalli PV, Waizenegger IC, Bunz F, Langer S, Speicher MR, et al. Securin is required for chromosomal stability in human cells. Cell 105: 44557. 45. Pfleghaar K, Heubes S, Cox J, Stemmann O, Speicher MR Securin is not required for chromosomal stability in human cells. PLoS Biol 3: 2127134. 46. Jones KT Meiosis in oocytes: predisposition to aneuploidy and its increased incidence with age. Human Reprod. Update. pp 16. 47. Pacchierotti F, Adler ID, 23472002 Eichenlaub-Ritter U, Mailhes JB Gender effects on the incidence of aneuploidy in mammalian germ cells. Environ Res 104: 469. 48. Wilhelm D, Koopman P The makings of maleness: towards an integrated view of male sexual development. Nat Rev Genet 7: 62031. 49. Berta P, Hawkins JR, Sinclair AH, Taylor A, Griffiths BL, et al. Genetic evidence equating SRY and the testis-determining factor. Nature 348: 44850. 50. Ginsburg M, Snow MH, McLaren A Primordial germ cells in the mouse Isoimperatorin customer reviews embryo during gastrulation. Development 110: 52128. 10 April 2011 | Volume 6 | Issue 4 | e18763 Cell-Type Specific Expression of a Dominant Negative PKA Mutation in Mice Brandon S. Willis, Colleen M. Niswender, Thomas Su, Paul S. Amieux, G. Stanley McKnight Department of Pharmacology, University of Washington, Seattle, Washington, United States of America Abstract We employed the Cre recombinase/loxP system to create a mouse line in which PKA activity can be inhibited in any celltype that expresses Cre recombinase. The mouse line carries a mutant Prkar1a allele encoding a glycine to aspartate substitution at position 324 in the carboxy-terminal cAMP-binding domain. This mutation produces a dominant negative RIa regulatory subunit and leads to inhibition of PKA activity. Insertion of a loxP-flanked neomycin cassette in the intron preceding the site B mutation prevents expression of the mutant RIaB allele until Cre-mediated excision of the cassette occurs. Embryonic stem cells expressing RIaB demonstrated a reduction in PKA activity and inhibition of cAMPresponsive gene expression. Mice expressing RIaB in hepatocytes exhibited reduced PKA activity, normal fasting induced gene expression, and enhanced glucose disposal. Activation of the RIaB allele in vivo provides a novel system for the analysis of PKA function in physiology. Citation: Willis BS, Niswender CM, Su T, Amieux PS, McKnight GS Cell-Type Specific Expression of a Dominant Negative PKA Mutation in Mice. PLoS ONE 6: e18772. doi:10.1371/journal.pone.0018772 Editor: Lin Mei, Medical College of Ge 172: 84760. 39. Hirano T, Funahashi SI, Uemura T, Yanagida M Isolation and characterization of Schizosaccharomyces pombe cut mutants that block nuclear division but not cytokinesis. EMBO J 5: 2973979. 40. Uzawa S, Samejima I, Hirano T, Tanaka K, Yanagida M The fission yeast cut1+ gene regulates spindle pole body duplication and has homology to the budding yeast ESP1 gene. Cell 62: 91325. 41. Stratmann R, Lehner 
CF Separation of sister chromatids in mitosis requires the Drosophila pimples product, a protein degraded after the metaphase/anaphase transition. Cell 84: 255. 42. Mei J, Huang X, Zhang P Securin is not required for cellular viability, but is required for normal growth of mouse embryonic fibroblasts. 17135238 Curr Biol 11: 1197201. 43. Wang Z, Yu R, Melmed S Mice lacking pituitary tumor transforming gene show testicular and splenic hypoplasia, thymic hyperplasia, thrombocytopenia, aberrant cell cycle progression, and premature centromere division. Mol Endocrinol 15: 1870879. 44. Jallepalli PV, Waizenegger IC, Bunz F, Langer S, Speicher MR, et al. Securin is required for chromosomal stability in human cells. Cell 105: 44557. 45. Pfleghaar K, Heubes S, Cox J, Stemmann O, Speicher MR Securin is not required for chromosomal stability in human cells. PLoS Biol 3: 2127134. 46. Jones KT Meiosis in oocytes: predisposition to aneuploidy and its increased incidence with age. Human Reprod. Update. pp 16. 47. Pacchierotti F, Adler ID, Eichenlaub-Ritter U, Mailhes JB Gender effects on the incidence of aneuploidy in mammalian germ cells. Environ Res 104: 469. 48. Wilhelm D, Koopman P The makings of maleness: towards an integrated view of male sexual development. Nat Rev Genet 7: 62031. 49. Berta P, Hawkins JR, Sinclair AH, Taylor A, Griffiths BL, et al. Genetic evidence equating SRY and the testis-determining factor. Nature 348: 44850. 50. Ginsburg M, Snow MH, McLaren A Primordial germ cells in the mouse embryo during gastrulation. Development 110: 52128. 10 April 2011 | Volume 6 | Issue 4 | e18763 Cell-Type Specific Expression of a Dominant Negative PKA Mutation in Mice Brandon S. Willis, Colleen M. Niswender, Thomas Su, Paul S. Amieux, G. Stanley McKnight Department of Pharmacology, University of Washington, Seattle, Washington, United States of America Abstract We employed the Cre recombinase/loxP system to create a mouse line in which PKA activity can be inhibited in any celltype that expresses Cre recombinase. The mouse line carries a mutant Prkar1a allele encoding a glycine to aspartate substitution at position 324 in the carboxy-terminal cAMP-binding domain. This mutation produces a dominant negative RIa regulatory subunit and leads to inhibition of PKA activity. Insertion of a loxP-flanked neomycin cassette in the intron preceding the site B mutation prevents expression of the mutant RIaB allele until Cre-mediated excision of the cassette occurs. Embryonic stem cells expressing RIaB demonstrated a reduction in PKA activity and inhibition of cAMPresponsive gene expression. Mice expressing RIaB in hepatocytes exhibited reduced PKA activity, normal fasting induced gene expression, and enhanced glucose disposal. Activation of the RIaB allele in vivo provides a novel system for the analysis of PKA function in physiology. Citation: Willis BS, Niswender CM, Su T, Amieux PS, McKnight GS Cell-Type Specific Expression of a Dominant Negative PKA Mutation in Mice. PLoS ONE 6: e18772. doi:10.1371/journal.pone.0018772 Editor: Lin Mei, Medical College of Ge
