From owner-dros@hgmp.mrc.ac.uk Fri Apr 2 15:17:42 2004 Return-Path: X-Original-To: dros-outgoing Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id 977067D126 for ; Fri, 2 Apr 2004 15:17:42 +0100 (BST) X-Original-To: dros-list@hgmp.mrc.ac.uk Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id B0F357D0F6 for ; Fri, 2 Apr 2004 15:17:41 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6020) id 9A10B7D0DF; Fri, 2 Apr 2004 15:17:38 +0100 (BST) X-Original-To: bionet-drosophila@net.bio.net Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 9A66E7D0DE for ; Fri, 2 Apr 2004 08:23:04 +0100 (BST) Received: from mailbox4.ucsd.edu (mailbox4.ucsd.edu [132.239.1.56]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id BC85F7D147 for ; Fri, 2 Apr 2004 08:23:00 +0100 (BST) Received: from proxy.google.com (proxy.google.com [216.239.37.5]) by mailbox4.ucsd.edu (8.13.0.Alpha0/8.13.0.Alpha0) with ESMTP id i327MtM1032273 for ; Thu, 1 Apr 2004 23:22:56 -0800 (PST) Received: from by Google Production Router with ESMTP id i327Mq3u004933 for ; Thu, 1 Apr 2004 23:22:52 -0800 Received: (from news@localhost) by Google Production with id i327MqHl011171 for bionet-drosophila@moderators.isc.org; Thu, 1 Apr 2004 23:22:52 -0800 To: bionet-drosophila@moderators.isc.org From: maneesh@drunkenbastards.com (maneesh) Newsgroups: bionet.drosophila Subject: simple questions on oligo delivery to embryo Organization: http://groups.google.com Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit X-Trace: posting.google.com 1080890572 11170 127.0.0.1 (2 Apr 2004 07:22:52 GMT) X-Complaints-To: groups-abuse@google.com NNTP-Posting-Date: Fri, 2 Apr 2004 07:22:52 +0000 (UTC) X-Spamscanner: mailbox4.ucsd.edu (v1.4 Mar 2 2004 11:47:57, 0.0/5.0 2.63) X-MailScanner: PASSED (v1.2.8 20948 i327MtM1032273 mailbox4.ucsd.edu) Date: Fri, 2 Apr 2004 15:17:38 +0100 (BST) Sender: owner-dros@hgmp.mrc.ac.uk Precedence: bulk [please post reply as my email box is filled with spam] Hi all, I am working on a qualifying proposal (outside the field of research which I am participitating in, I will not actually perform the experiment but it should be all be sound) and I have come up with some silly idea of studying miRNA:miRNA interactions in drosophila (see Lai et. al in RNA(2004) 10:171). I am not going in hoping to blow the minds of my commitee, just come up with something sensible. I am particularly interested in studying the potential regulation of miR-4 and miR-79 by miR-9a, miR-9b, miR-9c. miR-4 and miR-79 have complmentary Brd box binding domains, and the miR9-a and miR-9c have perfectly conserved Brd box on the miRNA* strand. I haven't been able to trace through all the predicted targets at the miRNA registry, but most appear to be invovled in development and Lai says that the Brd box consensus sequence implicates these genes in the Notch signalling pathway. I believe that defective Notch pathways have either embryonic lethal phenotypes or flies that keep folded wings and which die quickly. So in proposing to understand the potential roles of these 5 miRNAs and how they relate to each other, I will have to be able to selectively express and silence them and (verify then look for) Notch pathway defective phenotypes. A recent issue of PLOS describes 2'-OMe RNA oligos as selective inhibitors of miRNA function, which appear to work well in cell culture. Now one thing I embarassingly wasn't thinking about was, that I am working with embryos and not cell culture (doh! I have done mammalian cell culture and transfection before). I can see a few ways to deliver miRNA constructs to cells, but I'll have some problems acheiving silencing in trying to deliver chemically modified oligos to embryos...won't I? If anyone can suggest an idea to me, or suggest some other way to shut off the miRNAs that I am interested in, please let me know or point me to a reference/tutorial....if you see any gaping holes in my basic proposal please hit me with the cluestick. Many thanks in advance [please remember to post ;)] .