Enhanced Golic+: Gene targeting with 100% recovery in<i>Drosophila</i>male germ cells

Chen, Huimin; Yao, Xiaohao; Ren, Qingzhong; Chang, Chuan-Chie; Liu, Lingyu; Lee, Tzumin

doi:10.1101/681874

2019

DOI: 10.1101/681874

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Enhanced Golic+: Gene targeting with 100% recovery inDrosophilamale germ cells

Huimin Chen

Xiaohao Yao

Qingzhong Ren

et al.

Abstract: 18 19

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CAMIO: a transgenic CRISPR pipeline to create diverse targeted genome deletions in Drosophila

Chen

Marqués

Sugino

et al. 2020

Nucleic Acids Research

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The genome is the blueprint for an organism. Interrogating the genome, especially locating critical cis-regulatory elements, requires deletion analysis. This is conventionally performed using synthetic constructs, making it cumbersome and non-physiological. Thus, we created Cas9-mediated Arrayed Mutagenesis of Individual Offspring (CAMIO) to achieve comprehensive analysis of a targeted region of native DNA. CAMIO utilizes CRISPR that is spatially restricted to generate independent deletions in the intact Drosophila genome. Controlled by recombination, a single guide RNA is stochastically chosen from a set targeting a specific DNA region. Combining two sets increases variability, leading to either indels at 1–2 target sites or inter-target deletions. Cas9 restriction to male germ cells elicits autonomous double-strand-break repair, consequently creating offspring with diverse mutations. Thus, from a single population cross, we can obtain a deletion matrix covering a large expanse of DNA at both coarse and fine resolution. We demonstrate the ease and power of CAMIO by mapping 5′UTR sequences crucial for chinmo's post-transcriptional regulation.

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CAMIO: a transgenic CRISPR pipeline to create diverse targeted genome deletions in Drosophila

Chen

Marqués

Sugino

et al. 2020

Nucleic Acids Research

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Enhanced Golic+: Gene targeting with 100% recovery inDrosophilamale germ cells

Abstract: 18 19

Cited by 1 publication

References 20 publications

CAMIO: a transgenic CRISPR pipeline to create diverse targeted genome deletions in Drosophila

CAMIO: a transgenic CRISPR pipeline to create diverse targeted genome deletions in Drosophila

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