Title:
METHOD FOR INTRODUCING NUCLEIC ACID INTO CELLS AND APPLICATION THEREOF
Document Type and Number:
WIPO Patent Application WO/2024/043202
Kind Code:
A1
Abstract:
The present invention addresses the problem of providing: a method for introducing a nucleic acid into cells, wherein the method can efficiently introduce a high-concentration nucleic acid into high-density cells as a complex of the nucleic acid and a cationic polymer; a method for producing cells with nucleic acid introduced and a method for producing a useful substance; and a composition containing a cationic polymer and a nucleic acid, and a cell culture containing the composition and cells. The present invention provides a method for introducing a nucleic acid into cells comprising: (a) a complex formation step that mixes a cationic polymer and a nucleic acid to obtain a solution of a complex of the cationic polymer and nucleic acid, wherein the concentration of the nucleic acid in the complex formation step is 100-1000 μg/mL and the electrical conductivity of the mixed complex solution is 1.0-7.0 mS/cm; and (b) a transfection step that adds the complex solution obtained in step (a) to a cell suspension.
Inventors:
NISHINO MASAFUMI (JP)
SATO RYOTA (JP)
MURAGUCHI TAICHI (JP)
MORIYA KAI (JP)
SATO RYOTA (JP)
MURAGUCHI TAICHI (JP)
MORIYA KAI (JP)
Application Number:
PCT/JP2023/029939
Publication Date:
February 29, 2024
Filing Date:
August 21, 2023
Export Citation:
Assignee:
FUJIFILM CORP (JP)
International Classes:
C12N15/88; C12N5/071; C12N5/10; C12N15/85; C12N15/86
Foreign References:
| JP2011024493A | 2011-02-10 | |||
| JP2021106580A | 2021-07-29 |
Other References:
SUÁREZ-PATIÑO SANDRA FERNANDA; MANCINI RENATO ASTRAY; PEREIRA CARLOS AUGUSTO; SUAZO CLAUDIO ALBERTO TORRES; MENDONÇA RONALDO ZUCAT: "Transient expression of rabies virus glycoprotein (RVGP) inDrosophila melanogasterSchneider 2 (S2) cells", JOURNAL OF BIOTECHNOLOGY, ELSEVIER, AMSTERDAM NL, vol. 192, 7 July 2014 (2014-07-07), Amsterdam NL , pages 255 - 262, XP029104801, ISSN: 0168-1656, DOI: 10.1016/j.jbiotec.2014.05.027
MANCINELLI SARA, TURCATO ANDREA, KISSLINGER ANNAMARIA, BONGIOVANNI ANTONELLA, ZAZZU VALERIA, LANATI ANTONELLA, LIGUORI GIOVANNA LU: "Design of transfections: Implementation of design of experiments for cell transfection fine tuning", BIOTECHNOLOGY AND BIOENGINEERING, JOHN WILEY, HOBOKEN, USA, vol. 118, no. 11, 1 November 2021 (2021-11-01), Hoboken, USA, pages 4488 - 4502, XP093144310, ISSN: 0006-3592, DOI: 10.1002/bit.27918
ALATORRE-MEDA MANUEL, TABOADA PABLO, KRAJEWSKA BARBARA, WILLEMEIT MARKUS, DEML ALEXANDER, KLÖSEL ROLAND, RODRÍGUEZ JULIO R.: "DNA−Poly(diallyldimethylammonium chloride) Complexation and Transfection Efficiency", JOURNAL OF PHYSICAL CHEMISTRY PART B, AMERICAN CHEMICAL SOCIETY, US, vol. 114, no. 29, 29 July 2010 (2010-07-29), US , pages 9356 - 9366, XP093144313, ISSN: 1520-6106, DOI: 10.1021/jp1016856
MANCINELLI SARA, TURCATO ANDREA, KISSLINGER ANNAMARIA, BONGIOVANNI ANTONELLA, ZAZZU VALERIA, LANATI ANTONELLA, LIGUORI GIOVANNA LU: "Design of transfections: Implementation of design of experiments for cell transfection fine tuning", BIOTECHNOLOGY AND BIOENGINEERING, JOHN WILEY, HOBOKEN, USA, vol. 118, no. 11, 1 November 2021 (2021-11-01), Hoboken, USA, pages 4488 - 4502, XP093144310, ISSN: 0006-3592, DOI: 10.1002/bit.27918
ALATORRE-MEDA MANUEL, TABOADA PABLO, KRAJEWSKA BARBARA, WILLEMEIT MARKUS, DEML ALEXANDER, KLÖSEL ROLAND, RODRÍGUEZ JULIO R.: "DNA−Poly(diallyldimethylammonium chloride) Complexation and Transfection Efficiency", JOURNAL OF PHYSICAL CHEMISTRY PART B, AMERICAN CHEMICAL SOCIETY, US, vol. 114, no. 29, 29 July 2010 (2010-07-29), US , pages 9356 - 9366, XP093144313, ISSN: 1520-6106, DOI: 10.1021/jp1016856
Attorney, Agent or Firm:
SIKS & CO. (JP)
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