{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T07:24:18Z","timestamp":1777965858243,"version":"3.51.4"},"reference-count":32,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5339","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1997,10,31]]},"abstract":"<jats:p>\n            To create mice expressing exclusively human sickle hemoglobin (HbS), transgenic mice expressing human \u03b1-, \u03b3-, and \u03b2\n            <jats:sup>S<\/jats:sup>\n            -globin were generated and bred with knockout mice that had deletions of the murine \u03b1- and \u03b2-globin genes. These sickle cell mice have the major features (irreversibly sickled red cells, anemia, multiorgan pathology) found in humans with sickle cell disease and, as such, represent a useful in vivo system to accelerate the development of improved therapies for this common genetic disease.\n          <\/jats:p>","DOI":"10.1126\/science.278.5339.876","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:50:55Z","timestamp":1027763455000},"page":"876-878","source":"Crossref","is-referenced-by-count":413,"title":["Transgenic Knockout Mice with Exclusively Human Sickle Hemoglobin and Sickle Cell Disease"],"prefix":"10.1126","volume":"278","author":[{"given":"Chris","family":"Pa\u0301szty","sequence":"first","affiliation":[{"name":"C. Pa\u0301szty and N. Mohandas, Human Genome Center and Department of Subcellular Structure, Lawrence Berkeley National Laboratory, 1 Cyclotron Road (MS 74-157), University of California, Berkeley, CA 94720, USA."},{"name":"C. M. Brion, M. E. Stevens, E. M. Rubin, Human Genome Center, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA."},{"name":"E. Manci, Centralized Pathology Unit for Sickle Cell Disease, University of South Alabama Doctors Hospital, Mobile, AL 36604, USA."},{"name":"H. E. Witkowska, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catherine M.","family":"Brion","sequence":"additional","affiliation":[{"name":"C. Pa\u0301szty and N. Mohandas, Human Genome Center and Department of Subcellular Structure, Lawrence Berkeley National Laboratory, 1 Cyclotron Road (MS 74-157), University of California, Berkeley, CA 94720, USA."},{"name":"C. M. Brion, M. E. Stevens, E. M. Rubin, Human Genome Center, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA."},{"name":"E. Manci, Centralized Pathology Unit for Sickle Cell Disease, University of South Alabama Doctors Hospital, Mobile, AL 36604, USA."},{"name":"H. E. Witkowska, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elizabeth","family":"Manci","sequence":"additional","affiliation":[{"name":"C. Pa\u0301szty and N. Mohandas, Human Genome Center and Department of Subcellular Structure, Lawrence Berkeley National Laboratory, 1 Cyclotron Road (MS 74-157), University of California, Berkeley, CA 94720, USA."},{"name":"C. M. Brion, M. E. Stevens, E. M. Rubin, Human Genome Center, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA."},{"name":"E. Manci, Centralized Pathology Unit for Sickle Cell Disease, University of South Alabama Doctors Hospital, Mobile, AL 36604, USA."},{"name":"H. E. Witkowska, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H. Ewa","family":"Witkowska","sequence":"additional","affiliation":[{"name":"C. Pa\u0301szty and N. Mohandas, Human Genome Center and Department of Subcellular Structure, Lawrence Berkeley National Laboratory, 1 Cyclotron Road (MS 74-157), University of California, Berkeley, CA 94720, USA."},{"name":"C. M. Brion, M. E. Stevens, E. M. Rubin, Human Genome Center, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA."},{"name":"E. Manci, Centralized Pathology Unit for Sickle Cell Disease, University of South Alabama Doctors Hospital, Mobile, AL 36604, USA."},{"name":"H. E. Witkowska, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mary E.","family":"Stevens","sequence":"additional","affiliation":[{"name":"C. Pa\u0301szty and N. Mohandas, Human Genome Center and Department of Subcellular Structure, Lawrence Berkeley National Laboratory, 1 Cyclotron Road (MS 74-157), University of California, Berkeley, CA 94720, USA."},{"name":"C. M. Brion, M. E. Stevens, E. M. Rubin, Human Genome Center, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA."},{"name":"E. Manci, Centralized Pathology Unit for Sickle Cell Disease, University of South Alabama Doctors Hospital, Mobile, AL 36604, USA."},{"name":"H. E. Witkowska, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Narla","family":"Mohandas","sequence":"additional","affiliation":[{"name":"C. Pa\u0301szty and N. Mohandas, Human Genome Center and Department of Subcellular Structure, Lawrence Berkeley National Laboratory, 1 Cyclotron Road (MS 74-157), University of California, Berkeley, CA 94720, USA."},{"name":"C. M. Brion, M. E. Stevens, E. M. Rubin, Human Genome Center, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA."},{"name":"E. Manci, Centralized Pathology Unit for Sickle Cell Disease, University of South Alabama Doctors Hospital, Mobile, AL 36604, USA."},{"name":"H. E. Witkowska, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Edward M.","family":"Rubin","sequence":"additional","affiliation":[{"name":"C. Pa\u0301szty and N. Mohandas, Human Genome Center and Department of Subcellular Structure, Lawrence Berkeley National Laboratory, 1 Cyclotron Road (MS 74-157), University of California, Berkeley, CA 94720, USA."},{"name":"C. M. Brion, M. E. Stevens, E. M. Rubin, Human Genome Center, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA."},{"name":"E. Manci, Centralized Pathology Unit for Sickle Cell Disease, University of South Alabama Doctors Hospital, Mobile, AL 36604, USA."},{"name":"H. E. Witkowska, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.110.2865.543"},{"key":"e_1_3_1_2_3","doi-asserted-by":"publisher","DOI":"10.1038\/178792a0"},{"key":"e_1_3_1_3_2","unstructured":"S. H. Embury R. P. Hebbel N. Mohandas M. H. Steinberg Eds. Sickle Cell Disease: Basic Principles and Clinical Practice (Raven Press New York 1994)."},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2154033"},{"key":"e_1_3_1_4_3","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1038\/343183a0","volume":"343","author":"Greaves D. R.","year":"1990","unstructured":"Greaves D. R., et al., Nature 343, 183 (1990);","journal-title":"Nature"},{"key":"e_1_3_1_4_4","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1991.tb04877.x"},{"key":"e_1_3_1_4_5","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1172\/JCI115041","volume":"87","author":"Rubin E. M.","year":"1991","unstructured":"Rubin E. M., et al., J. Clin. Invest. 87, 639 (1991);","journal-title":"J. Clin. Invest."},{"key":"e_1_3_1_4_6","doi-asserted-by":"crossref","first-page":"12150","DOI":"10.1073\/pnas.89.24.12150","volume":"89","author":"Fabry M. E.","year":"1992","unstructured":"Fabry M. E., Nagel R. L., Pachnis A., Suzuka S. M., Costantini F., Proc. Natl. Acad. Sci. U.S.A. 89, 12150 (1992);","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_4_7","unstructured":"; M. E. Fabry et al. ibid. p. 12155."},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"3189","DOI":"10.1182\/blood.V84.9.3189.3189","volume":"84","author":"Trudel M.","year":"1994","unstructured":"Trudel M., et al., Blood 84, 3189 (1994);","journal-title":"Blood"},{"key":"e_1_3_1_5_3","first-page":"2419","volume":"86","author":"Fabry M. E.","year":"1995","unstructured":"Fabry M. E., et al., ibid. 86, 2419 (1995).","journal-title":"ibid."},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","unstructured":"C. P\u00e1szty Curr. Opin. Hematol. 4 88 (1997).","DOI":"10.1097\/00062752-199704020-00003"},{"key":"e_1_3_1_7_2","unstructured":"Three DNA fragments were used to create transgenic mice: (i) 6.5-kb mini-LCR (Locus Control Region) ["},{"key":"e_1_3_1_7_3","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1038\/338352a0","volume":"338","author":"Talbot D.","year":"1989","unstructured":"Talbot D., et al., Nature 338, 352 (1989);","journal-title":"Nature"},{"key":"e_1_3_1_7_4","unstructured":"] (ii) 1.5-kb Pst I fragment containing the human \u03b1 1 -globin gene (iii) 39-kb Kpn I fragment containing the human fetal ( G \u03b3 A \u03b3) and adult (\u03b4 \u03b2 S ) globin genes in their normal genomic context ["},{"key":"e_1_3_1_7_5","doi-asserted-by":"crossref","first-page":"4894","DOI":"10.1073\/pnas.81.15.4894","volume":"81","author":"Collins F. S.","year":"1984","unstructured":"Collins F. S., Stoeckert C. J., Serjeant G. R., Forget B. G., Weissman S. M., Proc. Natl. Acad. Sci. U.S.A. 81, 4894 (1984)].","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_8_2","unstructured":"Globin chain compositions were determined by high-performance liquid chromatography (HPLC) on a Vydac C 4 column as described (8)."},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1038\/ng0995-33","volume":"11","author":"P\u00e1szty C.","year":"1995","unstructured":"P\u00e1szty C., et al., Nature Genet. 11, 33 (1995).","journal-title":"Nature Genet."},{"key":"e_1_3_1_10_2","doi-asserted-by":"crossref","first-page":"9259","DOI":"10.1073\/pnas.92.20.9259","volume":"92","author":"Ciavatta D. J.","year":"1995","unstructured":"Ciavatta D. J., Ryan T. M., Farmer S. C., Townes T. M., Proc. Natl. Acad. Sci. U.S.A. 92, 9259 (1995).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_11_2","unstructured":"Genotyping was as follows. Polymerase chain reaction (PCR) conditions: 0.5 \u03bcM each primer 200 \u03bcM deoxynucleotide triphosphate 1 unit of Taq polymerase 20-\u03bcl reaction volume; 94\u00b0C (3 min); 35 cycles of 94\u00b0C (30 s) 58\u00b0C (30 s) 72\u00b0C (1 min). Sickle transgene forward primer \u03b4 1 (5\u2032-GTATGGGAGAGGCTCCAACTC-3\u2032) reverse primer \u03b4 2 (5\u2032-TCTGCCCAAATCTTAGACAAAAC-3\u2032). \u03b4 1\u03b42 PCR product 480 base pairs (bp). Mouse \u03b1-globin genotypes (+\/\/+ Hba 0 \/\/+ Hba 0 \/\/ Hba 0 ); Hba 0 allele: forward primer Hba 0 I (5\u2032-ATAGATGGGTAGCCATTTAGATTCC-3\u2032) reverse primer Hba 0 II (5\u2032-CCGGGTTATAATTACCTCAGGTC-3\u2032). Hba 0 IHba 0 II PCR product 461 bp. Also used previously published \u03b1-globin multiplex PCR (8). Mouse \u03b2-globin genotypes (+\/\/+ Hbb 0 \/\/+ Hbb 0 \/\/ Hbb 0 ); Hbb 0 allele: forward primer Hbb 0 I (5\u2032-AGATGTTTTTTTCACATTCTTGAGC-3\u2032) reverse primer Hbb 0 II (5\u2032-AATGCCTGCTCTTTACTGAAGG-3\u2032). Hbb 0 IHbb 0 II PCR product 398 bp. Also used PCR for part of region deleted in the Hbb 0 allele: forward primer \u03b2 del1 (5\u2032-TTAGGTGGTCTTAAAACTTTTGTGG-3\u2032) reverse primer \u03b2 del2 (5\u2032-ACTGGCACAGAGCATTGTTATG-3\u2032). \u03b2 del1\u03b2del2 PCR was with Deep Vent (Exonuclease \u2212 ) instead of Taq. Genotypes of sickle cell mice were confirmed by cellulose acetate gel electrophoresis of hemoglobins (Helena Laboratories Beaumont TX) ["},{"key":"e_1_3_1_11_3","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1007\/BF00484723","volume":"16","author":"Whitney J. B.","year":"1978","unstructured":"Whitney J. B., Biochem. Genet. 16, 667 (1978)].","journal-title":"Biochem. Genet."},{"key":"e_1_3_1_12_2","unstructured":"Sickle cell mice are fertile; however females appear to have trouble feeding their pups and as a result many of the sickle cell mice we have produced thus far have come from sickle cell males bred with non\u2013sickle cell females [Tg(Hu-miniLCR\u03b1 1 G \u03b3 A \u03b3\u03b4\u03b2 S ) Hba 0 \/\/ Hba 0 Hbb 0 \/\/+]. Interestingly there was a fivefold increase in sickle cell newborn survival when sickle cell males were bred with non\u2013sickle cell females (number obtained\/number expected = 32\/122) compared with when non\u2013sickle cell males were bred with non\u2013sickle cell females (number obtained\/number expected = 4\/82). One possible explanation for this is the presence of segregating \u201csurvival\u201d alleles in the mixed genetic background (FVB\/N 129 DBA\/2 C57BL\/6 Black Swiss) of the breeding population. These alleles would be inherited at a higher frequency from sickle cell males than from non\u2013sickle cell males. Another possible explanation involves the presence of fewer healthy non\u2013sickle cell newborns in litters from sickle cell males. This might allow sickle cell newborns to compete more effectively for maternal attention and enable them to survive more readily. The following animal husbandry practices may influence the survival of sickle cell mice: Micro-Barrier Systems cages (17.8 \u00d7 27.9 cm) (Allentown Caging Allentown NJ) Bed-O'Cobs bedding (size 0.32 cm) (Andersons Industrial Products Maumee OH) Nestlets nesting material (Ancare Corp. Bellmore NY) and Purina 5008 food (PMI Inc.)."},{"key":"e_1_3_1_13_2","unstructured":"Globin chain synthesis was as described (8). Controls used were C57BL\/6 (1.0 \u00b1 0.02; n = 3) and Hbb 0 \/\/+ \u03b2-thalassemic mice (1.38 \u00b1 0.05; n = 3)."},{"key":"e_1_3_1_14_2","unstructured":"Blood was mixed (1:1) with a 2% sodium bisulfite solution ["},{"key":"e_1_3_1_14_3","first-page":"1082","volume":"33","author":"Daland G. A.","year":"1948","unstructured":"Daland G. A., Castle W. B., J. Lab. Clin. Med. 33, 1082 (1948);","journal-title":"J. Lab. Clin. Med."},{"key":"e_1_3_1_14_4","unstructured":"] and wet mounts were prepared immediately. After 15 min 80% of cells were completely sickled (60% of cells had a single domain or a few parallel domains 20% had multispiculated holly leaf morphology)."},{"key":"e_1_3_1_15_2","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1182\/blood.V61.5.899.899","volume":"61","author":"Clark M. R.","year":"1983","unstructured":"Clark M. R., Mohandas N., Shohet S. B., Blood 61, 899 (1983).","journal-title":"Blood"},{"key":"e_1_3_1_16_2","unstructured":"The increases (fold) in organ weights were calculated from organ weights expressed as a percentage of total body weight [sickle cell ( n = 4) versus wild type ( n = 5): kidneys 2.5% versus 1.3%; heart 1.0% versus 0.5%; spleen 6.5% versus 0.5%]."},{"key":"e_1_3_1_17_2","unstructured":"Histological analysis was performed on four sickle cell and two wild-type mice all 5 to 7 months of age."},{"key":"#cr-split#-e_1_3_1_18_2.1","unstructured":"P. M. Chauhan P. Kondlapoodi C. L. Natta in Pathology Annual S. C. Sommers and P. P. Rosen Eds. (Appleton-Century-Crofts Norwalk CT 1983) vol. 18 pp. 253-276"},{"key":"#cr-split#-e_1_3_1_18_2.2","doi-asserted-by":"crossref","unstructured":"D. Powars J. A. Weidman T. Odom-Maryon J. C. Niland C. Johnson Medicine 67 66 (1988).","DOI":"10.1097\/00005792-198801000-00005"},{"key":"e_1_3_1_19_2","unstructured":"We thank P. Cooper F. Kuypers B.-C. Lau M. Sorette B. Bookchin B. Mentzer J. Zhang P. Donohue J. Hanneman and K. Brinkley for their contribution to this work. Supported by National Heart Lung and Blood Institute (NHLBI) grants HL31579 and HL20985 to the Northern California Comprehensive Sickle Cell Center a Red Cell Program Project Grant (DK32094) from the National Institute of Diabetes Digestive and Kidney Diseases a NHLBI contract (NO1-HB-07086) to the Sickle Cell Disease Centralized Pathology Unit and an NIH Shared Instrumentation Grant (VGBioQ mass spectrometer). Care of experimental animals was in accordance with institutional guidelines. Informed consent was obtained for human blood samples."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.278.5339.876","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T05:34:11Z","timestamp":1705124051000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.278.5339.876"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1997,10,31]]},"references-count":32,"journal-issue":{"issue":"5339","published-print":{"date-parts":[[1997,10,31]]}},"alternative-id":["10.1126\/science.278.5339.876"],"URL":"https:\/\/doi.org\/10.1126\/science.278.5339.876","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1997,10,31]]}}}