MicroRNA Knock Down by Cholesterol-Conjugated Antisense Oligos in Mouse Organ Culture
Here, we detail a protocol to design and introduce sequence-specific cholesterol-conjugated antisense oligonucleotides into mouse organ culture. We review design principles for “antagomirs”, antisense oligos with a cholesterol-moiety modification at the 3′, and present an optimized method to apply them onto 3D cultured embryonic pancreas. The method offers an approach to study the developmental functions of individual miRNAs and to evaluate miRNA targets, which is significantly faster and simpler than comparable genetics-based approaches.
Key wordsAntagomir miRNA silencing Cholesterol conjugate Pancreas miRNA targets
We thank Judith Maggenheim and Yuval Dor (Hadassah Medical School, The Hebrew University, Jerusalem, Israel) for protocols and advice and Tal Melkman-Zehavi for comments on this manuscript. EH is the incumbent of the Helen and Milton A Kimmelman Career Development Chair. This work was supported by grants from Juvenile Diabetes Research Foundation (#99-2007-71), the EFSD/D-Cure Young Investigator award, the Israel Science Foundation, the Yeda-Sela Center for Basic Research and the Wolfson Family Charitable Trust.
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