Organ-targeted high-throughput in vivo biologics screen identifies materials for RNA delivery.
Organ-targeted high-throughput in vivo biologics screen identifies materials for RNA delivery.
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DOI：
10.1039/c4ib00150h
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发表时间：
2014-10
期刊：
影响因子：
--
通讯作者：
Yanik MF
中科院分区：
文献类型：
--
作者：
Chang TY;Shi P;Steinmeyer JD;Chatnuntawech I;Tillberg P;Love KT;Eimon PM;Anderson DG;Yanik MF
基于生物制剂的治疗包括蛋白质、DNA和RNA的输送，是目前最有希望的方法之一。然而，尽管可以很容易地合成生物制剂和递送载体的大型组合文库，但目前还没有方法使用动物模型在体内快速表征它们。在这里，我们展示了通过自动递送到器官发达的小型脊椎动物的目标组织中，对生物制品和递送工具进行高通量体内筛选。单个斑马鱼幼体使用微流控系统以阵列形式自动定位并固定在水凝胶液滴中，递送工具以几乎完美的重复性和精确度自动显微注射到目标器官。我们筛选了一个类脂递送载体的文库，以了解它们在中枢神经系统促进蛋白质编码RNA表达的能力。我们发现了对斑马鱼幼体和老鼠都有效的递送工具。我们的结果表明，与体外哺乳动物细胞培养试验相比，体内斑马鱼模型可以显著更好地预测哺乳动物的假阳性和假阴性。我们的筛选结果也提示了一定的结构-活性关系，这可能会被应用于设计新型的递送载体。
Therapies based on biologics involving delivery of proteins, DNA, and RNA are currently among the most promising approaches. However, although large combinatorial libraries of biologics and delivery vehicles can be readily synthesized, there are currently no means to rapidly characterize them in vivo using animal models. Here, we demonstrate high-throughput in vivo screening of biologics and delivery vehicles by automated delivery into target tissues of small vertebrates with developed organs. Individual zebrafish larvae are automatically oriented and immobilized within hydrogel droplets in an array format using a microfluidic system, and delivery vehicles are automatically microinjected to target organs with nearly perfect repeatability and precision. We screened a library of lipid-like delivery vehicles for their ability to facilitate the expression of protein-encoding RNAs in the central nervous system. We discovered delivery vehicles that are effective in both larval zebrafish and rats. Our results showed that the in vivo zebrafish model can be significantly more predictive of both false positives and false negatives in mammals than in vitro mammalian cell culture assays. Our screening results also suggest certain structure-activity relationship, which can potentially be applied to design novel delivery vehicles.
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影响因子：
46.9
作者：
Judge, AD;Sood, V;MacLachlan, I
通讯作者：
MacLachlan, I
影响因子：
12.4
作者：
Kariko, Katalin;Muramatsu, Hiromi;Welsh, Frank A.;Ludwig, Janos;Kato, Hiroki;Akira, Shizuo;Weissman, Drew
通讯作者：
Weissman, Drew
影响因子：
6.1
作者：
Chang TY;Pardo-Martin C;Allalou A;Wählby C;Yanik MF
通讯作者：
Yanik MF
影响因子：
48
作者：
Pardo-Martin, Carlos;Chang, Tsung-Yao;Koo, Bryan Kyo;Gilleland, Cody L.;Wasserman, Steven C.;Yanik, Mehmet Fatih
通讯作者：
Yanik, Mehmet Fatih
影响因子：
10.4
作者：
Blechinger SR;Warren JT Jr;Kuwada JY;Krone PH
通讯作者：
Krone PH