RT Journal Article SR Electronic T1 Various applications of TALEN- and CRISPR/Cas9-mediated homologous recombination to modify the Drosophila genome JF Biology Open JO Biology Open FD Company of Biologists SP BIO20147682 DO 10.1242/bio.20147682 A1 Yu, Zhongsheng A1 Chen, Hanqing A1 Liu, Jiyong A1 Zhang, Hongtao A1 Yan, Yan A1 Zhu, Nannan A1 Guo, Yawen A1 Yang, Bo A1 Chang, Yan A1 Dai, Fei A1 Liang, Xuehong A1 Chen, Yixu A1 Shen, Yan A1 Deng, Wu-Min A1 Chen, Jianming A1 Zhang, Bo A1 Li, Changqing A1 Jiao, Renjie YR 2014 UL http://bio.biologists.org/content/early/2014/03/05/bio.20147682.abstract AB Modifying the genomes of many organisms is becoming as easy as manipulating DNA in test tubes, which is made possible by two recently developed techniques based on either the customizable DNA binding protein, TALEN, or the CRISPR/Cas9 system. Here, we describe a series of efficient applications derived from these two technologies, in combination with various homologous donor DNA plasmids, to manipulate the Drosophila genome: (1) to precisely generate genomic deletions; (2) to make genomic replacement of a DNA fragment at single nucleotide resolution; and (3) to generate precise insertions to tag target proteins for tracing their endogenous expressions. For more convenient genomic manipulations, we established an easy-to-screen platform by knocking in a white marker through homologous recombination. Further, we provided a strategy to remove the unwanted duplications generated during the “ends-in” recombination process. Our results also indicate that TALEN and CRISPR/Cas9 had comparable efficiency in mediating genomic modifications through HDR (homology-directed repair); either TALEN or the CRISPR/Cas9 system could efficiently mediate in vivo replacement of DNA fragments of up to 5 kb in Drosophila, providing an ideal genetic tool for functional annotations of the Drosophila genome.