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Research Article

基于RNA（ADARs）同种型的腺苷脱氨酶对基因治疗遗传密码校正的比较活性

卷 19, 期 1, 2019

页: [31 - 39] 页: 9

弟呕挨: 10.2174/1566523218666181114122116

价格: $65

摘要

简介：作用于RNA（ADAR）酶家族的腺苷脱氨酶成员由双链RNA结合结构域（dsRBD）和将腺苷（A）转化为肌苷（I）的脱氨酶结构域（DD）组成，其作用如下： guanosine（G）在翻译期间。使用MS2系统，我们设计了ADAR1的DD，以将其定向到特定目标。这项工作的目的是比较ADAR1-DD的脱氨酶活性和ADAR2-DD的各种同种型。 材料和方法：我们测量了人工酶系统对Biacore™X100的结合亲和力。 ADAR通常靶向dsRNA，因此我们设计了与靶RNA互补的指导RNA，然后将指导序列融合到MS2茎环中。靶向EGFP的58个氨基酸（TGG）的突变的琥珀（TAG）终止密码子。在将这三种因子转染到HEK 293细胞中后，我们观察到各种强度的荧光信号。 结果：没有Alu盒的ADAR2-long产生的荧光信号比Alu-box的ADAR2长得多。使用另一种同种型，ADAR2-short，其在C-末端短81bp，荧光信号是不可检测的。 ADAR2-long-DD（E488Q）的单个氨基酸取代使得酶比野生型更具活性。荧光显微镜检查结果表明，ADAR1-DD比ADAR2-long-DD更活跃。 Western印迹和测序证实ADAR1-DD比任何其他DD更活跃。 结论：本研究提供的信息应有助于合理使用ADAR变异体进行遗传病的遗传修复和治疗。

关键词: 定点RNA编辑，基因治疗，ADAR-DD，dsRBD，TAG，MS2 RNA。

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[1]
Napoli C, Lemieux C, Jorgensen R. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 1990; 2(4): 279-89.
[2]
Jinek M, Chylinski K, Fonfara I, et al. A programmable dual-RNA-guided DNA endonuclease in adoptive bacterial immunity. Science 2012; 337: 816-22.
[3]
Montiel-González MF, Vallecillo-Viejo IC, Rosenthal JJC. An efficient system for selectively altering genetic information within mRNAs. Nucleic Acids Res 2016; 44(21): 1-12.
[4]
Nishikura K. Functions and regulation of RNA editing by ADAR deaminases. Annu Rev Biochem 2010; 79: 321-49.
[5]
Maas S, Rich A. Changing genetic information through RNA editing. BioEssays 2000; 22(9): 790-802.
[6]
Sun T, Bentolila S, Hanson MR. The unexpected diversity of plant organelle RNA editosomes. Trends Plant Sci 2016; 21(11): 962-73.
[7]
Wu B, Chen H, Shao J, et al. Identification of symmetrical RNA editing events in the mitochondria of salvia miltiorrhiza by strand-specific RNA sequencing. Sci Rep 2017; 7(1): 42250.
[8]
Maas S, Rich A, Nishikura K. A-to-I RNA editing: Recent news and residual mysteries. J Biol Chem 2003; 278(3): 1391-4.
[9]
Bass BL. RNA editing by adenosine deaminates that act on RNA. Annu Rev Biochem 2002; 71(I): 817-46.
[10]
Kim U, Wang Y, Sanford T, et al. Molecular cloning of cDNA for double-stranded RNA adenosine deaminase, a candidate enzyme for nuclear RNA editing. Proc Natl Acad Sci USA 1994; 91(24): 11457-61.
[11]
Keegan LP, Gallo A, O’Connell MA. The many roles of an RNA editor. Nat Rev Genet 2001; 2(11): 869-78.
[12]
Bass BL, Nishikura K, Keller W, et al. A standardized nomenclature for adenosine deaminases that act on RNA. RNA 1997; 3: 947-9.
[13]
Chen CX, Cho DS, Wang Q, et al. A third member of the RNA-specific adenosine deaminase gene family, ADAR3, contains both single- and double-stranded RNA binding domains. RNA 2000; 6(5): 755-67.
[14]
Woolf TM, Chase JM, Stinchcomb DT. Toward the therapeutic editing of mutated RNA sequences. Proc Natl Acad Sci 1995; 92(18): 8298-302.
[15]
Schneider MF, Wettengel J, Hoffmann PC, et al. Optimal guideRNAs for re-directing deaminase activity of hADAR1 and hADAR2 in trans. Nucleic Acids Res 2014; 42(10): 1-9.
[16]
Melcher T, Maas S, Herb A, et al. RED2, a brain-specific member of the RNA-specific adenosine deaminase family. J Biol Chem 1996; 271(50): 31795-8.
[17]
Rueter SM, Dawson TR, Emeson RB. Regulation of alternative splicing by RNA editing. Nature 1999; 399(5): 75-80.
[18]
Wettengel J, Reautschnig P, Geisler S, et al. Harnessing human ADAR2 for RNA repair - Recoding a PINK1 mutation rescues mitophagy. Nucleic Acids Res 2017; 45(5): 2797-808.
[19]
Montiel-Gonzalez MF, Vallecillo-Viejo I, Yudowski GA, et al. Correction of mutations within the cystic fibrosis transmembrane conductance regulator by site-directed RNA editing. Proc Natl Acad Sci 2013; 110(45): 18285-90.
[20]
Lai F, Chen CX, Carter KC, et al. Editing of glutamate receptor B subunit ion channel RNAs by four alternatively spliced DRADA2 double-stranded RNA adenosine deaminases. Mol Cell Biol 1997; 17(5): 2413-24.
[21]
Gerber A, O’Connell MA, Keller W. Two forms of human double-stranded RNA-specific editase 1 (hRED1) generated by the insertion of an Alu cassette. RNA 1997; 3: 453-63.
[22]
Macbeth MR, Lingam AT, Bass BL. Evidence for auto-inhibition by the N terminus of hADAR2 and activation by dsRNA binding. RNA 2004; 10(10): 1563-71.
[23]
Hanswillemenke A, Kuzdere T, Vogel P, et al. Site-directed RNA editing in vivo can be triggered by the light-driven assembly of an artificial riboprotein. J Am Chem Soc 2015; 137(50): 15875-81.
[24]
Vu LT, Nguyen TTK, Md Thoufic AA, et al. Chemical RNA editing for genetic restoration: The relationship between the structure and deamination efficiency of carboxyvinyldeoxyuridine oligodeoxynucleotides. Chem Biol Drug Des 2016; 87(4): 583-93.
[25]
Vu LT, Nguyen TTK, Alam S, et al. Changing blue fluorescent protein to green fluorescent protein using chemical RNA editing as a novel strategy in genetic restoration. Chem Biol Drug Des 2015; 86(5): 1242-52.
[26]
Cattaneo R, Schmid A, Eschle D, et al. Biased hypermuation and other genetic changes in defective measles viruses in human brain infections. Cell 1988; 55: 255-65.
[27]
Polson AG, Bass BL, Casey JL. RNA editing of hepatitis delta virus antigenome by dsRNA-adenosine deaminase. Nature 1996; 380: 454-45.
[28]
Azad TA, Bhakta S, Tsukahara T. Site-directed RNA editing by adenosine deaminase acting on RNA (ADAR1) for correction of the genetic code in gene therapy. Gene Ther 2017; 24: 779-86.
[29]
Keryer-Bibens C, Barreau C, Osborne HB. Tethering of proteins to RNAs by bacteriophage proteins. Biol Cell 2008; 100(2): 125-38.
[30]
Buxbaum AR, Singer RH. In the right place at the right time: Visualizing and understanding mRNA localization. Nat Rev Mol Cell Biol 2015; 16(2): 95-109.
[31]
Rinkevich FD, Schweitzer PA, Scott JG. Antisense sequencing improves the accuracy and precision of A-to-I editing measurements using the peak height ratio method. BMC Res Notes 2012; 5(1): 63.
[32]
Eggington JM, Greene T, Bass BL. Predicting sites of ADAR editing in double-stranded RNA. Nat Commun 2011; 2(5): 319.
[33]
Yang J, Yan R, Roy A, et al. The I-TASSER Suite: Protein structure and function prediction. Nat Methods 2015; 12(1): 7-8.
[34]
Roy A, Kucukural A, Zhang Y. I-TASSER: A unified platform for automated protein structure and function prediction. Nat Protoc 2010; 5(4): 725-38.
[35]
Zhang Y. I-TASSER server for protein 3D structure prediction. BMC Bioinformatics 2008; 9(1): 40.
[36]
Battey JND, Kopp J, Bordoli L, Read RJ. Automated server predictions in CASP7. Proteins 2007; 69(Suppl. 8): 68-82.
[37]
Moult J, Pedersen JT, Judson R, et al. A large-scale experiment to assess protein structure prediction methods. Proteins 1995; 23(3): ii-v.
[38]
Kuttan A, Bass BL. Mechanistic insights into editing-site specificity of ADARs. Proc Natl Acad Sci USA 2012; 109: E3295-304.
[39]
Macbeth MR, Schubert HL, Vandemark AP, Lingam AT, Hill CP, Bass BL. Inositol hexakisphosphate is bound in the ADAR2 core and required for RNA editing. Science 2005; 309: 2033-7.
[40]
Stefl R, Oberstrass FC, Hood JL, et al. The solution structure of the ADAR2 dsRBM-RNA complex reveals a sequence-specific readout of the minor groove. Cell 2010; 143(2): 225-37.
[41]
Matthews MM, Thomas JM, Zheng Y, et al. Structures of human ADAR2 bound to dsRNA reveal base-flipping mechanism and basis for site selectivity. Nat Struct Mol Biol 2016; 23(5): 426-33.
[42]
Nurpeisov V, Hurwitz SJ, Sharma PL. Fluorescent dye terminator sequencing methods for quantitative determination of replication fitness of human immunodeficiency virus type 1 containing the codon 74 and 184 mutations in reverse transcriptase. J Clin Microbiol 2003; 41(7): 3306-11.
[43]
Nakae A, Tanaka T, Miyake K, et al. Comparing methods of detection and quantitation of RNA editing of rat glycine receptor alpha3. Int J Biol Sci 2008; 4(6): 397-405.
[44]
Koeris M, Funke L, Shrestha J, et al. Modulation of ADAR1 editing activity by Z-RNA in vitro. Nucleic Acids Res 2005; 33(16): 5362-70.
[45]
Heep M, Mach P, Reautschnig P, et al. Applying human ADAR1p110 and ADAR1p150 for Site-Directed RNA editing-G/C substitution stabilizes guideRNAs against editing. Genes 2017; 8(1): 1-7.
[46]
Morales ME, White TB, Streva VA, et al. The contribution of Alu elements to mutagenic DNA double-strand break repair. PLoS Genet 2015; 11(3): 1-26.
[47]
Huang Y, Cao Y, Li J, et al. A survey on cellular RNA editing activity in response to Candida albicans infections. BMC Genomics 2018; 19(43): 31-41.
[48]
Phelps KJ, Tran K, Eifler T, et al. Recognition of duplex RNA by the deaminase domain of the RNA editing enzyme ADAR2. Nucleic Acids Res 2015; 43(2): 1123-32.
[49]
Desterro JMP, Keegan LP, Lafarga M, et al. Dynamic association of RNA-editing enzymes with the nucleolus. J Cell Sci 2003; 116(9): 1805-18.
[50]
Nie Y, Zhao Q, Su Y, et al. Subcellular distribution of adar1 isoforms is synergistically determined by three nuclear discrimination signals and a regulatory motif. J Biol Chem 2004; 279(13): 13249-55.
[51]
Strehblow A, Hallegger M, Jantsch MF. Nucleocytoplasmic distribution of human RNA- editing enzyme ADAR1 is modulated by double- stranded RNA-binding domains, a leucine-rich export signal, and a putative dimerization domain. Mol Biol Cell 2002; 13: 3822-35.
[52]
Vogel P, Moschref M, Li Q, et al. Efficient and precise editing of endogenous transcripts with SNAP-tagged ADARs. Nat Methods 2018; 15(7): 535-8.
[53]
Cox DBT, Gootenberg JS, Abudayyeh OO, et al. RNA editing with CRISPR-Cas13. Science 2017; 358(6366): 1019-27.

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