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<p>Angelman syndrome (AS) is a severe neurodevelopmental disorder most often caused by deletion of the maternally inherited <i>UBE3A</i> allele (<i>matUBE3A</i>). In neurons, a long non-coding antisense RNA (<i>Ube3a-ATS</i>) silences the paternally-inherited <i>UBE3A</i> allele (<i>patUBE3A</i>). Here, we find that delivery of a zinc finger nuclease (ZFN) pair targeted to 86 <i>Snord115</i> genes within <i>Ube3a-ATS</i> (ZFN17/18) using adeno-associated virus (AAV) can unsilence patUBE3A in primary neuron cultures and in the brain of a mouse model of AS for at least 9 weeks. The AAV vector genome integrated at ZFN17/18 on-target sites in cultured neurons and, as evidence of specificity, did not integrate at predicted off-target sites. AAV vectors carrying nickase and catalytically inactive ZFN17/18 variants failed to appreciably unsilence <i>patUbe3a</i> and did not integrate at on-target sites. In vivo, we observed significant knockdown of <i>Ube3a-ATS</i> in AS-model mice, resulting in some neurons reaching UBE3A levels like those of wild-type mice. ZFN17/18 did not downregulate <i>Snrpn</i>, <i>Snord116, or IPW</i> in vivo, genes that are associated with Prader-Willi syndrome. Overall, our findings demonstrate the potential use of multi-target ZFNs as therapeutics for AS.</p>

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Multi-targeting zinc finger nuclease vector unsilences paternal UBE3A in a mouse model of Angelman syndrome

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年11月25日

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  • Hannah O. Bazick,
  • Lucas M. James,
  • Bonnie Taylor-Blake,
  • Justin M. Wolter,
  • Mark J. Zylka

摘要

Angelman syndrome (AS) is a severe neurodevelopmental disorder most often caused by deletion of the maternally inherited UBE3A allele (matUBE3A). In neurons, a long non-coding antisense RNA (Ube3a-ATS) silences the paternally-inherited UBE3A allele (patUBE3A). Here, we find that delivery of a zinc finger nuclease (ZFN) pair targeted to 86 Snord115 genes within Ube3a-ATS (ZFN17/18) using adeno-associated virus (AAV) can unsilence patUBE3A in primary neuron cultures and in the brain of a mouse model of AS for at least 9 weeks. The AAV vector genome integrated at ZFN17/18 on-target sites in cultured neurons and, as evidence of specificity, did not integrate at predicted off-target sites. AAV vectors carrying nickase and catalytically inactive ZFN17/18 variants failed to appreciably unsilence patUbe3a and did not integrate at on-target sites. In vivo, we observed significant knockdown of Ube3a-ATS in AS-model mice, resulting in some neurons reaching UBE3A levels like those of wild-type mice. ZFN17/18 did not downregulate Snrpn, Snord116, or IPW in vivo, genes that are associated with Prader-Willi syndrome. Overall, our findings demonstrate the potential use of multi-target ZFNs as therapeutics for AS.