A Primer for Morpholino Use in Zebrafish

Bill, Brent R.; Petzold, Andrew M.; Clark, Karl J.; Schimmenti, Lisa A.; Ekker, Stephen C.

doi:10.1089/zeb.2008.0555

Cited by 397 publications

(301 citation statements)

References 49 publications

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“…S4). As the translation-blocking effects of MOs generally wear off within 4-5 d after injection, 32 these data suggest that autophagy during the first 4-5 d of development is essential for long-term fish viability.…”

Section: Autophagy Gene Knockdown Results In Increased Numbers Of Deamentioning

confidence: 73%

Autophagy is essential for cardiac morphogenesis during vertebrate development

et al. 2014

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Section: Autophagy Gene Knockdown Results In Increased Numbers Of Deamentioning

confidence: 73%

Autophagy is essential for cardiac morphogenesis during vertebrate development

et al. 2014

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“…45 Not surprisingly, PMOs have also proven to be effective anti-miRNA compounds in Zebrafish. Flynt et al 46 reported knockdown of miR-214 in Zebrafish, demonstrating a role for this miRNA in muscle cell development.…”

Section: Inhibiting Mirna Function With Synthetic Oligonucleotides Dementioning

confidence: 99%

Chemical modification and design of anti-miRNA oligonucleotides

2011

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Antisense techniques have been employed for over 30 years to suppress expression of target RNAs. Recently, microRNAs (miRNAs) have emerged as a new class of small, non-coding, regulatory RNA molecules that widely impact gene regulation, differentiation and disease states in both plants and animals. Antisense techniques that employ synthetic oligonucleotides have been used to study miRNA function and some of these compounds may have potential as novel drug candidates to intervene in diseases where miRNAs contribute to the underlying pathophysiology. Anti-miRNA oligonucleotides (AMOs) appear to work primarily through a steric blocking mechanism of action; these compounds are synthetic reverse complements that tightly bind and inactivate the miRNA. A variety of chemical modifications can be used to improve the performance and potency of AMOs. In general, modifications that confer nuclease stability and increase binding affinity improve AMO performance. Chemical modifications and/or certain structural features of the AMO may also facilitate invasion into the miRNA-induced silencing complex. In particular, it is essential that the AMO binds with high affinity to the miRNA 'seed region', which spans bases 2-8 from the 5¢-end of the miRNA.

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“…NudC MO-1 (5′-TCTTTCGTCATCGTCACCCATTTTC-3′) targeted part of the open reading frame of zebrafish NudC mRNA, the rescue construct was engineered to change the nucleotide sequence without altering the encoded protein. As NudC MO-2, cofilin 1 MO-1 and MO-2 separately target the 5′-UTR sequence of NudC and cofilin 1 mRNA, the open reading frames of these two genes were simply cloned into pCS107 vector for rescue experiments [43]. Capped and polyadenylated RNAs were synthesized with the SP6 mMessage mMachine kit (Ambion).…”

Section: Plasmids and Oligonucleotidesmentioning

confidence: 99%

NudC regulates actin dynamics and ciliogenesis by stabilizing cofilin 1

Zhang

et al. 2015

Cell Res

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Emerging data indicate that actin dynamics is associated with ciliogenesis. However, the underlying mechanism remains unclear. Here we find that nuclear distribution gene C (NudC), an Hsp90 co-chaperone, is required for actin organization and dynamics. Depletion of NudC promotes cilia elongation and increases the percentage of ciliated cells. Further results show that NudC binds to and stabilizes cofilin 1, a key regulator of actin dynamics. Knockdown of cofilin 1 also facilitates ciliogenesis. Moreover, depletion of either NudC or cofilin 1 causes similar ciliary defects in zebrafish, including curved body, pericardial edema and defective left-right asymmetry. Ectopic expression of cofilin 1 significantly reverses the phenotypes induced by NudC depletion in both cultured cells and zebrafish. Thus, our data suggest that NudC regulates actin cytoskeleton and ciliogenesis by stabilizing cofilin 1.

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A Primer for Morpholino Use in Zebrafish

Cited by 397 publications

References 49 publications

Autophagy is essential for cardiac morphogenesis during vertebrate development

Autophagy is essential for cardiac morphogenesis during vertebrate development

Chemical modification and design of anti-miRNA oligonucleotides

NudC regulates actin dynamics and ciliogenesis by stabilizing cofilin 1

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