You can also do plain old antisense......

Richard Gomer
Department of Biology
Texas A&M University
301 Old Main Drive
College Station, TX  77843-3474
979 458 5745
________________________________________
From: DICTY [[log in to unmask]] on behalf of Ricardo Escalante [[log in to unmask]]
Sent: Thursday, April 5, 2018 3:37 AM
To: [log in to unmask]
Subject: [DICTY] RNAi

Dear Dicty friends,
I have a couple of questions regarding the use of RNAi in Dicty, I am not sure if this has been a topic lately in this list. I am thinking about using a hairpin construct to knock-down a gene that seems to be essential. The idea is to use an extrachromosomal inducible vector to express a hairpin so I can modulate the level of downregulation of the gene. Thus the questions are: 1. have you ever used this strategy successfully? 2. Which is the appropriate size and structure of the hairpin? Have you ever used small constructs of 35 pb like the shRNAs used in mammalian cells? In the first methods book chapter 13 (Kulmann, Popova and Nellen) they describe really long hairpin RNAs. Do you really need such a long structures. Making a short construct would facilitate the cloning as the prize to order gBLOCKs is now quite affordable.


Any advise will be welcome
Best Wishes
Ricardo





Ricardo Escalante, PhD.
Instituto de Investigaciones Biomédicas Alberto Sols
Arturo Duperier 4
28029-Madrid
Spain
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