Genecare Research Institute Co., Ltd.

Agene Research Institute Co., Ltd.

Roche Vitamins Inc.

Other

Hoffmann-la Roche Inc.

Jsr Corporation

Dsm Nutritional Products Inc.

Yasuhiro Furuichi

Akira Shimamoto

Motoki Takagi

Tatsuo Hoshino

Tatsuya Kiyasu

Yoshie Nagahashi

Kazunobu Futami

Saori Kitao

Hitoshi Kimura

Ayumi Sato

Mikio Hikata

Masanori Watahiki

Eiji Ohara

Michael A Richardson

Keiko Kuribayashi

Yuko Shibata

Hiroko Funaki

Satoshi Kaneto

Double-stranded modified siRNA targeting a RecQL1 helicase gene includes a sense strand including the nucleotide sequence shown in SEQ ID NO: 1, and an antisense strand including the nucleotide sequence shown in SEQ ID NO: 2, wherein the sense strand includes 2'-substituted nucleotides at positions 2, 3, 4 and 13 in the nucleotide sequence shown in SEQ ID NO: 1, the sense strand further includes a 2&#x2032;-substituted nucleotide(s) at one or more positions selected from the group consisting of positions 12, 14, 17, 18 and 19 in the nucleotide sequence shown in SEQ ID NO: 1, wherein the position 2&#x2032; of the 2&#x2032;-substituted nucleotides is &#x2014;R, &#x2014;OR, &#x2014;ROR, &#x2014;ORORor &#x2014;ROROR, wherein Rrepresents a Calkyl group, and Rand Rindependently represent a Calkylene group.

Modified siRNA and pharmaceutical composition

The present invention revealed that by suppressing the expression of the WRN gene, the BLM gene, or the RecQ1 gene, which belong to the RecQ helicase family, apoptosis is induced in various cancer cells and their proliferation is suppressed. Compounds that suppress the expression of RecQ helicase family genes or the functions of RecQ helicase proteins are thought to have the activity of inducing apoptosis.

Apoptosis inducer for cancer cell

The present inventors discovered that inhibition of the expression of various genes associated with chromosome stabilization induces cancer cell-specific apoptosis and inhibits cell proliferation. Compounds that inhibit expression of a gene associated with chromosome stabilization or inhibit the function of a protein encoded by such a gene are thought to have cancer cell-specific apoptosis-inducing effects.

Cancer cell-specific apoptosis-inducing agents that target chromosome stabilization-associated genes

The present inventors discovered that although suppressing expression of the RecQ1 gene, a RecQ helicase family gene, shows suppressive effects on cell proliferation in cancer cells, such effects are not observed in human TIG3 cells (a normal diploid fibroblast cell line), which are normal cells. Hence, the present inventors discovered that siRNAs against RecQ1 gene have cancer cell-specific cell proliferation-suppressing effects that are mediated by suppression of the expression of said gene.

Cancer-cell-specific cell proliferation inhibitors

The RecQ4 helicase gene, belonging to the RecQ helicase gene family, is revealed herein to be the causative gene of Rothmund-Thomson syndrome. The present inventors found out that it is possible to diagnose Rothmund-Thomson syndrome by detecting mutation of this gene. Further, they uncovered that it is possible to treat patients of Rothmund-Thomson syndrome by utilizing normal RecQ4 helicase gene or proteins thereof.

Gene causative of Rothmund-Thomson syndrome and its gene product

Gene causative of Rothmund-Thomson syndrome and gene product

The present invention relates to the production process of biotin by fermentation using a genetically engineered microorganism, and DNA sequences and vectors to be used in such process.

Biotin biosynthetic genes having biotin synthase activity

Biotin biosynthetic genes

cDNA including the 5&prime;-terminal sequence of full-length mRNA with a cap structure is synthesized from a mRNA sample containing the full-length mRNA with the cap structure and non-full-length mRNA without any cap structure in mixture. At the first step, the phosphate group at 5&prime;-terminus of the non-full-length mRNA in the mRNA sample is removed. At the second step, the cap structure at the 5&prime;-terminus of the full-length mRNA in the mRNA sample is removed. At the third step, an oligoribonucleotide is ligated to the phosphate group at 5&prime;-terminus of mRNA generated through the first and second steps. At the fourth step, mRNA with the oligoribonucleotide ligated at the 5&prime;-terminus thereof at the third step is subjected to a reverse transcriptase process using a short-chain oligonucleotide capable of being annealed to an intermediate sequence within the mRNA as primer, to synthesize a first-strand cDNA.

Method for synthesizing cDNA from mRNA sample

A gene encoding: (a) a protein comprising an amino acid sequence of SEQ ID NO: 2; or (b) a protein having deletion, substitution or addition of at least one amino acid residue in the amino acid sequence of SEQ ID NO: 2, which has a helicase activity.

Human gene RecQ4 encoding helicase

Growing community of inventors

Kamakura, Japan

Yasuhiro Furuichi