TY - JOUR T1 - CAMIO for deletion analysis of endogenous DNA sequences in multicellular organisms JF - bioRxiv DO - 10.1101/658088 SP - 658088 AU - Hui-Min Chen AU - Jorge Garcia Marques AU - Ken Sugino AU - Dingjun Wei AU - Rosa Linda Miyares AU - Tzumin Lee Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/06/25/658088.abstract N2 - 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 high-throughput analysis of native DNA. CAMIO utilizes CRISPR that is spatially restricted to generate independent deletions. 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. ER -