RICE CRISPR: Rapidly increased cut ends by an exonuclease Cas9 fusion in zebrafish.

RICE CRISPR: Rapidly increased cut ends by an exonuclease Cas9 fusion in zebrafish.
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DOI:
10.1002/dvg.23044
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发表时间:
2017-08
期刊:
Genesis (New York, N.Y. : 2000)
影响因子:
--
通讯作者:
Wagner DS
Wagner DS
中科院分区:
其他
文献类型:
--
作者:
Clements TP;Tandon B;Lintel HA;McCarty JH;Wagner DS

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CRISPR-Cas9技术在不同生物体中的应用导致了基因组修饰工作的爆炸式增长。为了扩展应用程序的工具箱,我们创建了一个E。coli外切核酸酶I(sbcB)- Cas9融合体,其在斑马鱼胚胎中具有改变的酶活性。这种Cas9变体具有增加的突变效率,并且相对于野生型Cas9有利于更长的缺失。我们预计,这种变体将允许更有效地筛选F0表型和更大范围的基因组靶点突变,包括缺失调控区和在序列保守性差的转录单位(如lncRNA)中产生功能丧失突变,其中功能丧失可能需要更大的缺失。
Application of CRISPR-Cas9 technology in diverse organisms has resulted in an explosion of genome modification efforts. To expand the toolbox of applications, we have created an E. coli Exonuclease I (sbcB) - Cas9 fusion that has altered enzymatic activity in zebrafish embryos. This Cas9 variant has increased mutation efficiency and favors longer deletions relative to wild type Cas9. We anticipate that this variant will allow for more efficient screening for F0 phenotypes and mutation of a larger spectrum of genomic targets including deletion of regulatory regions and creating loss of function mutations in transcription units with poor sequence conservation such as lncRNAs where larger deletions may be required for loss of function.
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影响因子: --
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