Takara Bio Inc.

Katsuyuki Dodo

Junichi Mineno

Hideto Chono

Masanari Kitagawa

Tamaki Akashika

Naoki Saito

Tatsuji Enoki

Hikaru Takakura

Hirofumi Yoshioka

Yasuhiro Kawano

Shuhei Sakamoto

Koichi Inoue

Takahisa Tsukihara

Mayumi Shimomura

A method for quantification of an adeno-associated virus genome, including the steps of (a) preparing a composition containing a sample, at least one primer pair for use in amplification of only a nucleotide sequence contained in inverted terminal repeats of an adeno-associated virus, and an intercalating dye; (b) performing nucleic acid amplification reaction using the composition prepared in the step (a); and (c) detecting an amplified product obtained in the step (b). The present invention is especially useful in the fields of medicine, gene engineering, and biology.

Method for quantifying adeno-associated virus

The present invention provides a method for efficiently manufacturing a non-enveloped virus with high purity without laborious operation by cultivating cells having the ability to produce a non-enveloped virus and bringing the cells and an acidic solution into contact with each other. A non-enveloped virus vector manufactured by the method of the present invention and a composition having the non-enveloped viral vector as an active ingredient are very useful as gene transfer methods in the fields of basic research and clinical application gene therapy.

Method for manufacturing non-enveloped virus

The present invention provides an expression cassette comprising (1) an expression regulatory region having a sequence corresponding to a transcriptional activator-binding region and (2) a nucleic acid encoding a transcriptional activator which is capable of binding to the expression regulatory region, wherein the nucleic acid is operably linked to the expression regulatory region; a method for expressing a target product in a mammalian cell, a method for producing a cell expressing a target product, and a method for producing a target product in a mammalian cell using the expression cassette; and a kit comprising the expression cassette.

Expression cassette

The present invention addresses the problem of providing cell populations having a high proportion of CD4-positive naive T cells and/or CD4-positive central memory T cells, and a production method thereof. The present invention provides a production method for CD4-positive T cell populations which is characterized by using anti-CD3 antibodies, fibronectin fragments, and Interleukin-4. The method is characterized not only by the attainment of a cell group with a high proportion of CD4-positive naive T cells and/or CD4-positive central memory T cells, but also by a high bulk yield.

Production method for cell populations

Disclosed is a simple and highly efficient method for introducing a gene into a target cell using a retrovirus vector. The method comprises the steps of (a) placing a liquor containing a retrovirus vector having a foreign gene carried thereon into a bag for cell culture on which a retrovirus-binding substance has been immobilized, and incubating the liquor at a temperature lower than 25&#xb0; C. for 8-48 hours, thereby producing a culture bag having the retrovirus vector bound thereto, (b) adding a target cell to the culture bag that has been produced in step (a) and incubating the culture bag for 8 hours or less, and (c) flipping the culture bag upside down and incubating the culture bag. The gene introduction method is useful particularly in medicine, cell technology, gene technology, and embryologic technology.

Method for gene transfer

Disclosed is a method for introducing a gene into a target cell using a retrovirus vector, which is simple and highly efficient. Specifically disclosed is a method for introducing a gene into a target cell using a retrovirus vector, which comprises the steps of (a) placing a liquor containing a retrovirus vector having a foreign gene carried thereon into a culture vessel on which a retrovirus-binding substance has been immobilized, and incubating the liquor at a temperature lower than 25&#xb0; C. for 4 hours or longer, thereby producing a culture vessel having the retrovirus vector bound thereto, and (b) adding a target cell to the culture vessel that has been produced in step (a) and incubating the culture vessel. The gene introduction method is simple and highly efficient, and is useful particularly in the fields of medicine, cell technology, gene technology, embryologic technology and the like.

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Kusatsu, Japan

Katsuyuki Dodo