NDLI: Use of cDNA subtraction and RNA interference screens in combination reveals genes required for germ-line development in Caenorhabditis elegans.

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Use of cDNA subtraction and RNA interference screens in combination reveals genes required for germ-line development in Caenorhabditis elegans.

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Use of cDNA subtraction and RNA interference screens in combination reveals genes required for germ-line development in Caenorhabditis elegans.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Hanazawa, M. Ueno, N. Iino, Y. Mochii, M. Kohara, Y.
Description	Author Affiliation: Hanazawa M ( Molecular Genetics Research Laboratory, University of Tokyo, Tokyo 113-0033, Japan.);
Abstract	Caenorhabditis elegans is an ideal organism for the study of the molecular basis of fundamental biological processes such as germ-line development, especially because of availability of the whole genome sequence and applicability of the RNA interference (RNAi) technique. To identify genes involved in germ-line development, we produced subtracted cDNA pools either enriched for or deprived of the cDNAs from germ-line tissues. We then performed differential hybridization on the high-density cDNA grid, on which about 7,600 nonoverlapping expressed sequence tag (EST) clones were spotted, to identify a set of genes specifically expressed in the germ line. One hundred and sixty-eight clones were then tested with the RNAi technique. Of these, 15 clones showed sterility with a variety of defects in germ-line development. Seven of them led to the production of unfertilized eggs, because of defects in spermatogenesis (4 clones), or defects in the oocytes (3 clones). The other 8 clones led to failure of oogenesis. These failures were caused by germ-line proliferation defect (Glp phenotype), meiotic arrest, and defects in sperm--oocyte switch (Mog phenotype) among others. These results demonstrate the efficacy of the screening strategy using the EST library combined with the RNAi technique in C. elegans.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	15
Volume Number	98
Language	English
Publisher	National Academy of Sciences
Publisher Date	2001-07-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Caenorhabditis Elegans Genetics Gene Expression Genes, Helminth Oogenesis RNA, Antisense Spermatogenesis Animals DNA, Complementary DNA, Helminth Fertilization Infertility Nucleic Acid Hybridization Physiology Ovum Phenotype RNA RNA, Helminth RNA, Small Interfering Research Support, Non-U.S. Gov't Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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