Generation of outbred Ace2 knockout mice by RNA transfection of TALENs displaying colitis reminiscent pathophysiology and inflammation

详细信息    查看全文

• 作者：Chuxin Liu ; Liping Xiao ; Feida Li ; Huanhuan Zhang ; Qin Li ; Huan Liu…
• 关键词：Ace2 ; TALEN ; Gene knockout ; Colitis ; Animal model
• 刊名：Transgenic Research
• 出版年：2015
• 出版时间：June 2015
• 年：2015
• 卷：24
• 期：3
• 页码：433-446
• 全文大小：1,572 KB
• 参考文献：Bedell VM, Wang Y, Campbell JM, Poshusta TL, Starker CG, Krug RG 2nd, Tan W, Penheiter SG, Ma AC, Leung AY, Fahrenkrug SC, Carlson DF, Voytas DF, Clark KJ, Essner JJ, Ekker SC (2012) In vivo genome editing using a high-efficiency TALEN system. Nature 491:114-18View Article PubMed Central PubMed
  Brenner MK, Rill DR, Moen RC, Krance RA, Mirro J Jr, Anderson WF, Ihle JN (1993) Gene-marking to trace origin of relapse after autologous bone-marrow transplantation. Lancet 341:85-6View Article PubMed
  Byrnes JJ, Gross S, Ellard C, Connolly K, Donahue S, Picarella D (2009) Effects of the ACE2 inhibitor GL1001 on acute dextran sodium sulfate-induced colitis in mice. Inflamm Res 58:819-27View Article PubMed
  Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF (2011) Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res 39:e82View Article PubMed Central PubMed
  Conti P, Kempuraj D, Kandere K, Di Gioacchino M, Barbacane RC, Castellani ML, Felaco M, Boucher W, Letourneau R, Theoharides TC (2003) IL-10, an inflammatory/inhibitory cytokine, but not always. Immunol Lett 86:123-29View Article PubMed
  Crackower MA, Sarao R, Oudit GY, Yagil C, Kozieradzki I, Scanga SE, Oliveira-dos-Santos AJ, da Costa J, Zhang L, Pei Y, Scholey J, Ferrario CM, Manoukian AS, Chappell MC, Backx PH, Yagil Y, Penninger JM (2002) Angiotensin-converting enzyme 2 is an essential regulator of heart function. Nature 417:822-28View Article PubMed
  Deng ZB, Yuan AW, Luo W, Wang ND, Gong QL, Yu XL, Xue LQ (2013) Transmission of porcine circovirus type 2b (PCV2b) in Kunming mice. Acta Veterinaria Hung 61:234-43View Article
  Ding Q, Lee YK, Schaefer EA, Peters DT, Veres A, Kim K, Kuperwasser N, Motola DL, Meissner TB, Hendriks WT, Trevisan M, Gupta RM, Moisan A, Banks E, Friesen M, Schinzel RT, Xia F, Tang A, Xia Y, Figueroa E, Wann A, Ahfeldt T, Daheron L, Zhang F, Rubin LL, Peng LF, Chung RT, Musunuru K, Cowan CA (2013) A TALEN genome-editing system for generating human stem cell-based disease models. Cell Stem Cell 12:238-51View Article PubMed Central PubMed
  Donoghue M, Hsieh F, Baronas E, Godbout K, Gosselin M, Stagliano N, Donovan M, Woolf B, Robison K, Jeyaseelan R, Breitbart RE, Acton S (2000) A novel angiotensin-converting enzyme-related carboxypeptidase (ACE2) converts angiotensin I to angiotensin 1-. Circ Res 87:E1–E9View Article PubMed
  Doyle EL, Booher NJ, Standage DS, Voytas DF, Brendel VP, Vandyk JK, Bogdanove AJ (2012) TAL Effector-Nucleotide Targeter (TALE-NT) 2.0: tools for TAL effector design and target prediction. Nucleic Acids Res 40:W117–W122View Article PubMed Central PubMed
  Doyon Y, Vo TD, Mendel MC, Greenberg SG, Wang J, Xia DF, Miller JC, Urnov FD, Gregory PD, Holmes MC (2011) Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures. Nat Methods 8:74-9View Article PubMed
  Epinat JC, Arnould S, Chames P, Rochaix P, Desfontaines D, Puzin C, Patin A, Zanghellini A, Paques F, Lacroix E (2003) A novel engineered meganuclease induces homologous recombination in yeast and mammalian cells. Nucleic Acids Res 31:2952-962View Article PubMed Central PubMed
  Farrow PJ, Barrett LB, Stevenson M, Fisher KD, Finn J, Spice R, Allan MA, Berry M, Logan A, Seymour LW, Read ML (2006) Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons. Nucleic Acids Res 34:e80View Article PubMed Central PubMed
  Fischer AH, Jacobson KA, Rose J & Zeller R (2008) Hematoxylin and eosin staining of tissue and cell sections, CSH protocols 2008(5):pdb–prot4986. doi: 10.-101/?pdb.?prot4986
  Gurley SB, Allred A, Le TH, Griffiths R, Mao L, Philip N, Haystead TA, Donoghue M, Breitbart RE, Acton SL, Rockman HA, Coffman TM (2006) Altered blood pressure responses and normal cardiac phenotype in ACE2-null mice. J Clin Investig 116:2218-225View Article PubMed Central PubMed
  Guyton AC (1992) Kidneys and fluids in pressure regulation. Small volume but large pressure changes. Hypertension 19:I2–I8View Article PubMed
  Hashimoto T, Perlot T, Rehman A, Trichereau J, Ishiguro H, Paolino M, Sigl V, Hanada T, Hanada R, Lipinski S, Wild B, Camargo SM, Singer D, Richter A, Kuba K, Fukamizu A, Schreiber S, Clevers H, Verrey F, Rosenstiel P, Penninger JM (2012) ACE2 links amino acid malnutrition to microbial ecology and intestinal inflammation. Nature 487:477-81View Article PubMed
  Hockemeyer D, Wang H, Kiani S, Lai CS, Gao Q, Cassady JP, Cost GJ, Zhang L, Santiago Y, Miller JC, Zeitler B, Cherone JM, Meng X, Hinkley SJ, Rebar EJ, Gregory PD, Urnov FD, Jaenisch R (2011) Genetic engineering of human pluripotent cells using TALE nucleases. Nat Biotechnol 29:731-34View Article PubMed Central PubMed
  Izcue A, Powrie F (2012) Immunology: malnutrition promotes rogue bacteria. Nature 487:437-39View Article PubMed
  Karhausen J
• 作者单位：Chuxin Liu (1) (2)
  Liping Xiao (1) (2)
  Feida Li (1) (2)
  Huanhuan Zhang (1) (2)
  Qin Li (1) (2)
  Huan Liu (1) (2)
  Shujin Fu (1) (2)
  Chao Li (1) (2)
  Xingju Zhang (1) (2)
  Jun Wang (1)
  Nicklas H. Staunstrup (3)
  Yong Li (1) (2)
  Huanming Yang (1)
  
  1. BGI-Shenzhen, Beishan Industrial Zone, Yantian District, Shenzhen, 518083, China
  2. ShenZhen Engineering Laboratory for Genomics-Assisted Animal Breeding, BGI-Shenzhen, 8F Main Building, Beishan Industrial Zone, Yantian District, Shenzhen, 518083, China
  3. Department of Biomedicine, University of Aarhus, Wilhelm Meyers allé 1240, 8000, Aarhus C, Denmark
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Molecular Medicine
  Plant Genetics and Genomics
  Animal Genetics and Genomics
  Plant Sciences
  Human Genetics
  
• 出版者：Springer Netherlands
• ISSN：1573-9368

文摘

The angiotensin I converting enzyme 2 (ACE2) is a key factor in the maintenance of intestinal homeostasis. Dysregulation of homeostasis can lead to inflammation of the colon (colitis), which can cause life-threatening enfeeblement or even cancer. Animal models are valuable surrogates in deciphering the pathology behind such human conditions and for screening of putative therapeutic targets or treatment paradigms. However, development of disease models can be time-consuming and technical demanding, which might hamper their application-value. In this study, we genetically disrupted the mouse Ace2 gene by direct injection of in vitro transcribed mRNA coding for transcription activator-like effector nucleases (TALENs) into the cytoplasm of outbred Kunming mouse zygotes. Consequently, somatic mutations were induced with an efficiency of 57?%, of which 39?% were frameshift mutations. Moreover, all modifications were stably transferred during germline transmission. In Ace2-knockout male mice (Ace2 ?em class="EmphasisTypeItalic">/y ), we observed severe chemical induced colitis, characterized by considerable weight loss, diarrhea and a shortened colon length. Histologically, Ace2 mutations resulted in the infiltration of leukocytes and the overt damage of the intestinal mucosal barrier. In addition, we detected an increased expression of inflammatory cytokines in the colon tissue of Ace2 ?em class="EmphasisTypeItalic">/y mice. Collectively, the data indicate that high targeting efficiency and heritability can be achieved in an outbred mouse model by zygote injection of TALEN mRNA. Furthermore, the generated Ace2 ?em class="EmphasisTypeItalic">/y mice display phenotypic traits reminiscent of colitis and we anticipate that such mice can be of value in studies of the intestinal microbiome or fecal transplantation.