Abstract: The present disclosure provides a nucleic acid construct capable of regulating protein expression. Specifically, the present disclosure provides a nucleic acid construct including a 5? untranslated region (UTR) and a 3? UTR that are at least partially complementary to each other, wherein at least one of UTRs includes a non-complementary region and a complementary region with respect to the other UTR, and when all of the respective lengths of the non-complementary regions are of 1 base, a degree of complementarity of the 3? UTR to the 5? UTR is greater than 75%.

Abstract: The present disclosure provides plasmid DNA or a DNA/cationic substance complex containing a Rep and an ITR and exhibiting high AAV vector productivity. More specifically, in one exemplary aspect, the present disclosure provides a nucleic acid containing a Rep gene derived from an adeno-associated virus (AAV) of a first serotype, and an inverted terminal repeat (ITR) sequence derived from an AAV of a second serotype different from the first serotype and having a ?G value in a range of ?65 to ?95 kcal/mol.

Abstract: The present disclosure provides control of expression of a Rep gene by control of expression of a helper gene. In one aspect, the present disclosure provides a method for controlling the expression of a Rep gene, including a step of controlling the expression of a helper gene. In one embodiment, the helper gene includes a gene selected from adenovirus E1B, E2, E4 and VA. In one embodiment, the helper gene includes an adenovirus E1B gene. In one embodiment, the helper gene includes two or more helper genes related to the production of an adeno-associated virus (AAV)-derived construct, and optionally, the helper genes can be selected from E1A, E1B, E2, E4, and VA. In one embodiment, the helper gene includes a helper gene selected from E1A and E1B and a helper gene selected from E2, E4 and VA.

Abstract: The present disclosure provides production of a virus-derived construct plasmid. In one aspect, the present disclosure provides a plasmid encoding at least some elements of a virus-derived construct. In one embodiment, the present disclosure provides a plasmid containing at least some of nucleic acid sequences required to construct a virus-derived construct in an operably linked form. In one embodiment, the nucleic acid sequence required to construct a virus-derived construct includes (A) a nucleic acid sequence including a terminal repeat, (B) a nucleic acid sequence encoding a packaging factor, (C) a nucleic acid sequence encoding a structural protein, and (D) a nucleic acid sequence encoding a functional cofactor.

Abstract: In one aspect, the present disclosure provides a method for producing various virus-derived construct plasmids from starting plasmids (for example, virus-derived construct plasmids of the present disclosure).

Abstract: Provided are: a method for preparing a DNA unit composition in which the mol number of a plurality of DNA units is more uniform, and a method for creating concatenated DNA. The method for preparing a DNA unit composition has: a step for preparing solutions which contain a plurality of DNA units to which an added sequence is linked, and preparing a solution for each type of DNA unit and a step for, after preparing each of the solutions, measuring the concentration of the DNA unit in each of the solutions in a state where the added sequence is linked to the DNA unit, and on the basis of the results thereof, fractionating each of the solutions and making the mol number of the DNA unit in each of the solutions closer to being identical to one another.

Abstract: Provided are: a method for preparing a DNA unit composition in which the mol number of a plurality of DNA units is more uniform, and a method for creating concatenated DNA. The method for preparing a DNA unit composition has: a step for preparing solutions which contain a plurality of DNA units to which an added sequence is linked, and preparing a solution for each type of DNA unit; and a step for, after preparing each of the solutions, measuring the concentration of the DNA unit in each of the solutions in a state where the added sequence is linked to the DNA unit, and on the basis of the results thereof, fractionating each of the solutions and making the mol number of the DNA unit in each of the solutions closer to being identical to one another.