WO2014025056A1 - Procédé de cryptage et de décryptage à l'aide d'acide nucléique - Google Patents

Procédé de cryptage et de décryptage à l'aide d'acide nucléique Download PDF

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Publication number
WO2014025056A1
WO2014025056A1 PCT/JP2013/071752 JP2013071752W WO2014025056A1 WO 2014025056 A1 WO2014025056 A1 WO 2014025056A1 JP 2013071752 W JP2013071752 W JP 2013071752W WO 2014025056 A1 WO2014025056 A1 WO 2014025056A1
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WIPO (PCT)
Prior art keywords
nucleic acid
information
base
sequence
cell
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Application number
PCT/JP2013/071752
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English (en)
Inventor
Naoki Yamamoto
Yoshihisa Nakazawa
Nobuaki Suzuki
Original Assignee
Hitachi Zosen Corporation
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Application filed by Hitachi Zosen Corporation filed Critical Hitachi Zosen Corporation
Priority to JP2014557903A priority Critical patent/JP6175453B2/ja
Publication of WO2014025056A1 publication Critical patent/WO2014025056A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1065—Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids

Definitions

  • the present invention relates to a method for encrypting information and decrypting the information using a nucleic acid.
  • NPL 1 nucleic acid molecules
  • GMOs genetically modified organisms
  • NPL 1 also describes how to correct mutations in the coding nucleic acid molecules, which is introduced into a genetically modified organism, during cell division so that encrypted information can be correctly decrypted.
  • NPL l D. Hieder et al., BMC Bioinformatics, 2007, Vol. 8, No. 176, pp.1-11
  • the inventors have found that when the encryption of information is performed by encoding the information by a binary sequence and associating the binary digits "0" and "1" with either of a purine base or a pyrimidine base and the other base, respectively, such encrypted information can be correctly decrypted without being affected by transition mutation.
  • the present invention provides a method for encrypting information using a nucleic acid, which comprises the steps of:
  • nucleic acid introducing the nucleic acid into a cell
  • one of 0 and 1 constituting the binary sequence is associated with either of a purine base or a pyrimidine base, and the other is associated with the other base.
  • said step of introducing the nucleic acid into a cell comprises inserting the nucleic acid into a vector, and introducing the vector into the cell.
  • the present invention also provides a coding medium comprising the nucleic acid or the vector for use in the method.
  • the present invention further provides a method for decrypting the information that is encrypted according to the method mentioned above, which comprises the step of decoding the nucleic acid in the cell, thereby obtaining a base sequence.
  • said step of decoding comprises deciphering the nucleic acid using a decoding primer.
  • the method of decrypting further comprises converting the base sequence to a binary sequence associated therewith, and decrypting the information from the binary sequence.
  • encrypted information can be correctly decrypted without being affected by nucleic acid mutation during the division of the cell into which a coding nucleic acid is introduced.
  • the information can be encoded into a compact nucleic acid sequence, thereby reducing the affection due to the mutation, and reducing time and cost required to synthesize the nucleic acid.
  • bases can be flexibility selected to encode information.
  • the amount of GC can be controlled in a nucleic acid sequence, and there is no need to amplify a coding nucleic acid by PCR using any PCR polymerase adapted for GC-rich templates.
  • any specific base sequence can be easily integrated into a nucleic acid sequence.
  • Fig. 1 is a schematic diagram illustrating a method of encryption and decryption using a nucleic acid according to the present invention.
  • Fig. 1 is a schematic diagram illustrating a method of encryption and decryption using a nucleic acid according to the present invention.
  • base herein refers to either a purine base, namely "A (adenine)” or “G (guanine)", or a pyrimidine base, namely "C (cytosine)” or “T (thymine)", unless specified otherwise.
  • nucleic acid in a singular form herein refers to one or more nucleic acids, unless specified otherwise.
  • the encryption using a nucleic acid includes the steps of synthesizing a nucleic acid having a base sequence encoding information to be encrypted, and introducing the nucleic acid into a cell.
  • the information to be encrypted is, for example, a character string represented as "HITZ0001"by alphabet letters and numbers.
  • the information such as massage, including characters of alphabet letters and numbers
  • a binary sequence for example, on the basis of a code table showing the association of characters of alphabet letters and numbers with respective binary triplets (3 bits).
  • the alphabet letter "H” is encoded by a binary triplet "000”
  • the number "0” is encoded by a different binary triplet "100” as information signals.
  • the number of binary digits (the number of bits) for encoding or associating with each character in the information to be encrypted may be, for example, 3 bits, 4 bits, and any more number of bits. In one embodiment, when a minimum set of characters is a limited number of characters of alphabet letters, 3 bits may be used. In other embodiments, for example, in order to encode 26 characters of total alphabet letters, 52 characters of discriminated alphabet letters by capital letters and lower-case letters, or further including period, comma, colon, semicolon, and the like, " or to encode 10 characters of numbers or further including mathematical symbols, a larger number of bits may be used.
  • the information "HITZ0001" can be encoded by a binary sequence (which is also herein referred to a "coding binary sequence") of, for example, "000001010011100100100101" as the information signals.
  • the binary digit "0” is encoded by a purine base, namely "A” or “G”
  • the binary digit "1” is encoded by a pyrimidine base, namely "C” or “T”
  • a purine base namely "A” or “G”
  • a pyrimidine base namely "C” or “T”
  • a table showing the association of characters of alphabet letters and numbers with respective binary indications and/or the association of binary digits "0" and "1" with respective types of bases (which is also herein referred to as a "code table”) can be prepared.
  • Such a code table is preferably kept in secret.
  • the binary digit "0" may be encoded by a pyrimidine base, namely "C” or “T”
  • the binary digit "1” may be encoded by a purine base, namely "A” or "G”.
  • the binary sequence "000001010011100100100101” is mechanically converted to the base sequence "AGAGACATGGCCCAATGGCAGTAC” (SEQ ID No. 1), for example according to the predetermined algorithm with conversion means such as computer program.
  • the information "HITZ0001" to be encrypted is converted to the base sequence "AGAGACATGGCCCAATGGCAGTAC" (SEQ ID No. l). Therefore, the information is associated with or encoded by a nucleic acid which is also herein referred to as a "coding nucleic acid”.
  • the coding nucleic acid sequence may further include additional base sequences representing the start and end points of the encoded information.
  • the additional base sequence may be any base sequence, and examples thereof may include a base sequence associating with the binary digits "111" in the case of 3 bits.
  • the coding nucleic acid is synthesized using any procedure commonly used.
  • a DNA fragment having the above-mentioned base sequence is chemically synthesized using a DNA synthesizer, and subsequently amplified by PCR or the like until a desired amount thereof is obtained.
  • Such a DNA fragment may optionally include structural or functional sequences, including a restriction enzyme recognition sequence adapted for insertion into a vector, and a promoter sequence (cis-arrangement) for regulating DNA expression, in addition to the nucleic acid encoding the information to be encrypted.
  • structural or functional sequences including a restriction enzyme recognition sequence adapted for insertion into a vector, and a promoter sequence (cis-arrangement) for regulating DNA expression, in addition to the nucleic acid encoding the information to be encrypted.
  • the coding nucleic acid is introduced into a cell.
  • the coding nucleic acid may be inserted into a vector for introducing into a cell.
  • the coding nucleic acid may be introduced directly into a cell.
  • plasmids examples thereof include plasmids, phages, and phagemids.
  • plasmids include pUC'based plasmid, pSClOl-based plasmid, and Ti plasmid such as pBI121.
  • phages examples thereof include ⁇ -based phage and Ml3-based phage. Insertion of a nucleic acid fragment into a vector may be performed by any procedure commonly used.
  • a coding nucleic acid and vector as described above are provided as a coding medium.
  • a coding nucleic acid may be bound, immobilized or fixed to a medium to label the medium using any procedure commonly used.
  • the medium include genetic recombinant DNA, papers, nylon membranes, beads, slides, silicon substrates, and resins.
  • cell there is no particular limitation on the cell, and examples thereof include cells derived from plants, animals, and microorganisms, and preferably genetically modified plants.
  • the procedure for introducing a vector or a nucleic acid into a cell there is no particular limitation on the procedure for introducing a vector or a nucleic acid into a cell, and the procedure is appropriately selected depending on cells. Examples thereof include introduction of a plasmid into a competent cell; phage infection! and electroporation introducing a vector or a nucleic acid via a pore electrically formed through a cell membrane.
  • the coding nucleic acid may be amplified and retained with the vector in the cell (in the case of a plasmid, etc.), or may be transferred from the vector to a chromosome in the cell and retained on the chromosome (in the case of phage, etc.).
  • the decryption from a nucleic acid including the encrypted information includes the step of decoding the nucleic acid in the cell.
  • the coding nucleic acid which is introduced and maintained in the cell, may be decoded.
  • the coding nucleic acid may be isolated from the cell, and then subjected to sequencing, or the coding base sequence may be sequenced directly from the cell.
  • a decoding primer is used.
  • the base sequence of the decoding primer may be designed based on the base sequence encoding the information (i.e., the base sequence of the coding nucleic acid, or the coding nucleic acid sequence), or may be designed based on the base sequence of a portion of the vector.
  • a pair of the decoding primers may be designed based on 5' and 3' flanking sequences of the coding nucleic acid sequence, respectively.
  • the 5' and 3' flankin sequences may be respective sequences having tens of bases in a vector or a DNA fragment as mentioned above, adjacent to the 5' and 3' ends of the coding nucleic acid sequence.
  • the 5' and 3' flanking sequences may include any structural or functional sequence as mentioned above.
  • either of the decoding primers may be designed based on the base sequence of the coding nucleic acid, and the other may be designed based on the 5' or 3' flanking sequence.
  • the procedure for isolating the coding nucleic acid from the cell and examples thereof include nucleic acid amplification such as PCR.
  • a base sequence of "AGAGACATGGCCCAATGGCAGTAC” (SEQ ID No. 1) can be obtained.
  • the conversion of the base sequence "AGAGACATGGCCCAATGGCAGTAC” (SEQ ID No. 1) to a binary sequence is mechanically performed with predetermined algorithm, e.g.
  • the binary sequence "000001010011100100100101" is converted to the information "HITZ0001" on the basis of the code table shown in Fig. 1.
  • the encrypted information is decrypted.
  • the encrypted information can be correctly decrypted without being affected by the mutation.
  • encrypted information can be correctly decrypted without being affected by nucleic acid mutation during the division of the cell into which a coding nucleic acid is introduced. Furthermore, information can be encoded into a compact nucleic acid sequence, thereby reducing the affection due to the mutation, and reducing time and cost required to synthesize the nucleic acid. Moreover, bases can be flexibility selected to encode information. Thus, the amount of GC can be controlled in a nucleic acid sequence, and there is no need to amplify a coding nucleic acid by PCR using any PCR polymerase adapted for GC-rich templates. Furthermore, any specific base sequence can be easily integrated into a nucleic acid sequence.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
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  • Wood Science & Technology (AREA)
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  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
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  • Crystallography & Structural Chemistry (AREA)
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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un procédé permettant le décryptage correct d'une information cryptée en rendant un acide nucléique code introduit dans une cellule moins sensible à une affection en raison d'une mutation ayant lieu pendant la division cellulaire. L'invention concerne un procédé de cryptage utilisant un acide nucléique, comprenant les étapes consistant à : synthétiser un acide nucléique ayant une séquence de bases associée à l'information à crypter; et introduire l'acide nucléique dans une cellule; l'information étant codée par une séquence binaire, et l'un de 0 et 1 constituant la séquence binaire étant associé soit à une base purine soit à une base pyrimidine, et l'autre étant associée à l'autre base.
PCT/JP2013/071752 2012-08-07 2013-08-06 Procédé de cryptage et de décryptage à l'aide d'acide nucléique WO2014025056A1 (fr)

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JP2014557903A JP6175453B2 (ja) 2012-08-07 2013-08-06 核酸を用いる暗号化および復号化方法

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Cited By (3)

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TWI673604B (zh) * 2017-06-14 2019-10-01 俄羅斯聯邦商蘭迪佳德有限責任公司 信息編碼和信息解碼的方法
CN113380322A (zh) * 2021-06-25 2021-09-10 倍生生物科技(深圳)有限公司 人工核酸序列水印编码系统、水印字符串及编码和解码方法
CN114374504A (zh) * 2021-08-16 2022-04-19 中电长城网际系统应用有限公司 数据加密方法、解密方法、装置、服务器

Families Citing this family (1)

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JP7107956B2 (ja) * 2016-11-16 2022-07-27 カタログ テクノロジーズ， インコーポレイテッド 核酸ベースのデータ記憶のためのシステム

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI673604B (zh) * 2017-06-14 2019-10-01 俄羅斯聯邦商蘭迪佳德有限責任公司 信息編碼和信息解碼的方法
CN113380322A (zh) * 2021-06-25 2021-09-10 倍生生物科技(深圳)有限公司 人工核酸序列水印编码系统、水印字符串及编码和解码方法
CN113380322B (zh) * 2021-06-25 2023-10-24 倍生生物科技(深圳)有限公司 人工核酸序列水印编码系统、水印字符串及编码和解码方法
CN114374504A (zh) * 2021-08-16 2022-04-19 中电长城网际系统应用有限公司 数据加密方法、解密方法、装置、服务器

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JP6175453B2 (ja) 2017-08-02

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