Unknown

Dataset Information

0

An expanded toolkit for gene tagging based on MiMIC and scarless CRISPR tagging in Drosophila.


ABSTRACT: We generated two new genetic tools to efficiently tag genes in Drosophila. The first, Double Header (DH) utilizes intronic MiMIC/CRIMIC insertions to generate artificial exons for GFP mediated protein trapping or T2A-GAL4 gene trapping in vivo based on Cre recombinase to avoid embryo injections. DH significantly increases integration efficiency compared to previous strategies and faithfully reports the expression pattern of genes and proteins. The second technique targets genes lacking coding introns using a two-step cassette exchange. First, we replace the endogenous gene with an excisable compact dominant marker using CRISPR making a null allele. Second, the insertion is replaced with a protein::tag cassette. This sequential manipulation allows the generation of numerous tagged alleles or insertion of other DNA fragments that facilitates multiple downstream applications. Both techniques allow precise gene manipulation and facilitate detection of gene expression, protein localization and assessment of protein function, as well as numerous other applications.

SUBMITTER: Li-Kroeger D 

PROVIDER: S-EPMC6095692 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature

altmetric image

Publications

An expanded toolkit for gene tagging based on MiMIC and scarless CRISPR tagging in <i>Drosophila</i>.

Li-Kroeger David D   Kanca Oguz O   Lee Pei-Tseng PT   Cowan Sierra S   Lee Michael T MT   Jaiswal Manish M   Salazar Jose Luis JL   He Yuchun Y   Zuo Zhongyuan Z   Bellen Hugo J HJ  

eLife 20180809


We generated two new genetic tools to efficiently tag genes in <i>Drosophila</i>. The first, Double Header (DH) utilizes intronic MiMIC/CRIMIC insertions to generate artificial exons for GFP mediated protein trapping or T2A-GAL4 gene trapping in vivo based on Cre recombinase to avoid embryo injections. DH significantly increases integration efficiency compared to previous strategies and faithfully reports the expression pattern of genes and proteins. The second technique targets genes lacking co  ...[more]

Similar Datasets

| S-EPMC9239680 | biostudies-literature
| S-EPMC4499919 | biostudies-literature
| S-EPMC7670108 | biostudies-literature
| S-EPMC5354236 | biostudies-literature
| S-EPMC4313374 | biostudies-literature
| S-EPMC10015468 | biostudies-literature
| S-EPMC4703899 | biostudies-other
| S-EPMC6622542 | biostudies-literature
| S-EPMC5063382 | biostudies-literature
| S-EPMC6456373 | biostudies-literature