ExBoX - a simple Boolean exclusion strategy to drive expression in neurons.
ExBoX - a simple Boolean exclusion strategy to drive expression in neurons.
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DOI：
10.1242/jcs.257212
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发表时间：
2021-10-15
影响因子：
4
通讯作者：
Riccomagno MM
中科院分区：
文献类型：
--
作者：
Ubina T;Vahedi-Hunter T;Agnew-Svoboda W;Wong W;Gupta A;Santhakumar V;Riccomagno MM
现代单细胞生物学的出现揭示了曾经被认为更同质的细胞群体惊人的分子多样性。这种新认识的复杂性使得交叉遗传学方法对于在功能水平上理解和探索细胞异质性是必不可少的。在这里，我们建立在已有知识的基础上，开发一种基于腺相关病毒(AAV)的简单方法，通过布尔排除逻辑(AND)定义特定的细胞亚群。这种布尔排斥表达(Exbox)系统编码被特定重组酶(Cre或FlpO)启动而被另一重组酶关闭的目的基因。Exbox允许在只表达激活重组酶的细胞中对感兴趣的基因进行特定转录，但在同时表达两种重组酶的细胞中不允许。我们展示了Exbox系统在体外和体内严格调控荧光报告基因表达的能力，并通过在小鼠大脑中实现各种病毒传递的编码序列的表达，进一步证明了该系统的适应性。这一简单的策略将扩展可用于在各种系统中进行细胞和时间特异性基因表达的分子工具包。摘要：一种新的基于AAV的交叉方法的产生，以定义和定位特定的细胞亚群在时间和空间通过表达由布尔排除(Exbox)系统。
The advent of modern single-cell biology has revealed the striking molecular diversity of cell populations once thought to be more homogeneous. This newly appreciated complexity has made intersectional genetic approaches essential to understanding and probing cellular heterogeneity at the functional level. Here, we build on previous knowledge to develop a simple adeno-associated virus (AAV)-based approach to define specific subpopulations of cells by Boolean exclusion logic (AND NOT). This expression by Boolean exclusion (ExBoX) system encodes for a gene of interest that is turned on by a particular recombinase (Cre or FlpO) and turned off by another. ExBoX allows for the specific transcription of a gene of interest in cells expressing only the activating recombinase, but not in cells expressing both. We show the ability of the ExBoX system to tightly regulate expression of fluorescent reporters in vitro and in vivo, and further demonstrate the adaptability of the system by achieving expression of a variety of virally delivered coding sequences in the mouse brain. This simple strategy will expand the molecular toolkit available for cell- and time-specific gene expression in a variety of systems. Summary: The generation of a novel AAV-based intersectional approach to define and target specific subpopulations of cells in time and space via a Expression by Boolean Exclusion (ExBoX) system.
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有条件和可逆基因敲除一步的一代。
影响因子：
48
作者：
Andersson-Rolf A;Mustata RC;Merenda A;Kim J;Perera S;Grego T;Andrews K;Tremble K;Silva JC;Fink J;Skarnes WC;Koo BK
通讯作者：
Koo BK
小鼠大脑皮层 GABA 神经元组合靶向的策略和工具
影响因子：
16.2
作者：
He M;Tucciarone J;Lee S;Nigro MJ;Kim Y;Levine JM;Kelly SM;Krugikov I;Wu P;Chen Y;Gong L;Hou Y;Osten P;Rudy B;Huang ZJ
通讯作者：
Huang ZJ
DOI：
10.1073/pnas.1507125112
发表时间：
2015-06-09
影响因子：
11.1
作者：
Darmanis S;Sloan SA;Zhang Y;Enge M;Caneda C;Shuer LM;Hayden Gephart MG;Barres BA;Quake SR
通讯作者：
Quake SR
影响因子：
64.5
作者：
Daigle TL;Madisen L;Hage TA;Valley MT;Knoblich U;Larsen RS;Takeno MM;Huang L;Gu H;Larsen R;Mills M;Bosma-Moody A;Siverts LA;Walker M;Graybuck LT;Yao Z;Fong O;Nguyen TN;Garren E;Lenz GH;Chavarha M;Pendergraft J;Harrington J;Hirokawa KE;Harris JA;Nicovich PR;McGraw MJ;Ollerenshaw DR;Smith KA;Baker CA;Ting JT;Sunkin SM;Lecoq J;Lin MZ;Boyden ES;Murphy GJ;da Costa NM;Waters J;Li L;Tasic B;Zeng H
通讯作者：
Zeng H
影响因子：
16.2
作者：
Farago, AF;Awatramani, RB;Dymecki, SM
通讯作者：
Dymecki, SM