Polyplex Formation of Calf Thymus DNA with Branched and Linear Polyethyleneimine

Jungho Han; Seog K. Kim; Tae-Sub Cho; Jae-Cheol Lee; Hyun Sook Joung

Macromolecular Research, Vol.12, No.5, 501-506, October, 2004

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Polyplex Formation of Calf Thymus DNA with Branched and Linear Polyethyleneimine

Jungho Han, Seog K. Kim¹, Tae-Sub Cho^{1, †}, Jae-Cheol Lee², and Hyun Sook Joung³

KT&G Central Research Institute, 302 Shinseong, Yuseong, Daejeon 305-805, Korea
¹Department of Chemistry, Yeungnam University, 214-1 Dae, Kyoungsan, Gyoungbuk 712-749, Korea
²Department of Bio-Food Science and Technology, Samcheok National University, Samcheok, Gangwon 245-711, Korea
³Department of Food and Nutrition, Keimyung College, 700 Shindang, Daegu 704-703, Korea

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We have investigated the spectral properties of DNA, including its electric absorption, circular and linear dichroism (CD and LD), and fluorescence emission, in the DNA-linear polyethyleneimine (LPEI) and DNAbranched polyethyleneimine (BPEI) complexes at various polymer concentrations. The spectral properties of both complexes are similar. We observed a relatively moderate change in the absorption and CD spectra at low amine/DNA phosphate (N/P) ratios (< 0.5), followed by a drastic collapse within the N/P range from 0.8 and 1.0. The absorption and CD spectra recovered as the N/P ratio increased to ca. 1.2. In contrast, the LD and emission of ethidium intercalated between the DNA bases decreased almost linearly at N/P ratios between 0.0 and 1.0. These spectra never recovered at higher N/P ratios. We believe that the moderate changes in the spectrum at low N/P ratios occurred because of electrostatic interactions between DNA and BPEI, while the collapsed spectra at N/P ratios between 0.5 and 1.5 occurred because of condensation/aggregation of the DNA. Considering the structure of the polymers, we suggest that the secondary amino group of LPEI and all three amino groups of BPEI are equally involved in DNA condensation.

Keywords:DNA;spectroscopy;polyplex;polyethyleneimine;gene carrier

[References]

Chirila TV, Rakoczy PE, Garrett KL, Lou X, Constable IJ, Biomaterials, 23, 321 (2002)
Kircheis R, Wightman L, Wagner E, Adv. Drug Deliv. Rev., 53, 341 (2001)
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Tang MX, Szoka FC, Gene Ther., 4, 823 (1997)
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Boussif O,Ogris M, Steinlein P, Kursa M, Mechtler K, Kircheis R, Wagner E, Gene Ther., 5, 1425 (1998)
Boussif O, Zanta MA, Behr JP, Gene Ther., 3, 1074 (1996)
Boussif O, Lezoulach F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP, Proc. Natl. Acad. Sci. USA, 92, 7297 (1995)
Ogris M, Brunner S, Schuller S, Kircheis R, Wagner E, Gene Ther., 6, 595 (1999)
Kircheis R, Wightman L, Schreiber A, Robitza B, Rossler V, Kursa M, Gene Ther., 8, 28 (2001)
Wightman L, Kircheis R, Rossler V, Carotta R, Ruzicka R, Kursa M, Wagner E, J. Gene Med., 3, 362 (2001)
Kok SJ< Evertsen R, Velthorst NH, Th UA, Brinkman, Gooijer C, Anal. Chim. Acta, 405(35), 1 (2000)
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Chen W, Tomalia DA, Thomas JL, Macromol, 33, 9169 (2000)
Ottaviani MF, Furini F, Casini A, Turro NJ, Jockusch S, Tomalia DA, Messoori L, Macromol, 33, 7842 (2000)
Moret I, Peris JE, Guillem VM, Benet M, Revert F, Dasi F, Crespo A, Alino SF, J. Control. Release, 76, 169 (2001)
Han J, Kim SK, Cho TS, Lee JC, Macromol. Res., 12(3), 276 (2004)
Shiro K, Kazuhisa H, Masazumi T, Takeo S, Macromol., 20, 1496 (1987)
Norden B, Seth S, Appl. Spectrosc., 39, 647 (1985)
Norden B, Kubista M, Kurucsev T, O. Rev. Biophys., 25, 51 (1992)
Norden B, Kurucsev T, J. Mol. Recognit., 7, 141 (1994)
Pelta J, Livolant F, Sikorav JL, J. Biol. Chem., 271, 5656 (1996)
Raspaud E, Olvera de la Cruz M, Sikorav JL, Livolant F, Biophys. J., 74, 381 (1998)
Bloomfield VA, Biopolymers, 44, 269 (1994)

[Referenced By]

[1] Chirila TV, Rakoczy PE, Garrett KL, Lou X, Constable IJ, Biomaterials, 23, 321 (2002)

[2] Kircheis R, Wightman L, Wagner E, Adv. Drug Deliv. Rev., 53, 341 (2001)

[3] De Smedt SC, Demeester J, Hennink WE, Pharm. Res., 17, 113 (2000)

[4] Zauner W, Ogris M, Wagner E, Adv. Drug Deliv. Rev., 30, 97 (1998)

[5] Tang MX, Szoka FC, Gene Ther., 4, 823 (1997)

[6] Dunlap DD, Maggi A, Soria MR, Monaco L, Nucleic Acids Res., 25, 3095 (1997)

[7] Boussif O,Ogris M, Steinlein P, Kursa M, Mechtler K, Kircheis R, Wagner E, Gene Ther., 5, 1425 (1998)

[8] Boussif O, Zanta MA, Behr JP, Gene Ther., 3, 1074 (1996)

[9] Boussif O, Lezoulach F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP, Proc. Natl. Acad. Sci. USA, 92, 7297 (1995)

[10] Ogris M, Brunner S, Schuller S, Kircheis R, Wagner E, Gene Ther., 6, 595 (1999)

[11] Kircheis R, Wightman L, Schreiber A, Robitza B, Rossler V, Kursa M, Gene Ther., 8, 28 (2001)

[12] Wightman L, Kircheis R, Rossler V, Carotta R, Ruzicka R, Kursa M, Wagner E, J. Gene Med., 3, 362 (2001)

[13] Kok SJ< Evertsen R, Velthorst NH, Th UA, Brinkman, Gooijer C, Anal. Chim. Acta, 405(35), 1 (2000)

[14] Von Harpe A, Petersen H, Li Y, Kissel T, J. Control. Release, 69(1), 309 (2000)

[15] Bronich TK, Nguyen HK, Eisenberg A, Kabanov AV, J. Am. Chem. Soc., 122(35), 8339 (2000)

[16] Chen W, Turro NJ, Tomalia DA, Langmuir, 16(1), 15 (2000)

[17] Chen W, Tomalia DA, Thomas JL, Macromol, 33, 9169 (2000)

[18] Ottaviani MF, Furini F, Casini A, Turro NJ, Jockusch S, Tomalia DA, Messoori L, Macromol, 33, 7842 (2000)

[19] Moret I, Peris JE, Guillem VM, Benet M, Revert F, Dasi F, Crespo A, Alino SF, J. Control. Release, 76, 169 (2001)

[20] Han J, Kim SK, Cho TS, Lee JC, Macromol. Res., 12(3), 276 (2004)

[21] Shiro K, Kazuhisa H, Masazumi T, Takeo S, Macromol., 20, 1496 (1987)

[22] Norden B, Seth S, Appl. Spectrosc., 39, 647 (1985)

[23] Norden B, Kubista M, Kurucsev T, O. Rev. Biophys., 25, 51 (1992)

[24] Norden B, Kurucsev T, J. Mol. Recognit., 7, 141 (1994)

[25] Pelta J, Livolant F, Sikorav JL, J. Biol. Chem., 271, 5656 (1996)

[26] Raspaud E, Olvera de la Cruz M, Sikorav JL, Livolant F, Biophys. J., 74, 381 (1998)

[27] Bloomfield VA, Biopolymers, 44, 269 (1994)