Unknown

Dataset Information

0

Conditionals by inversion provide a universal method for the generation of conditional alleles.


ABSTRACT: Conditional mutagenesis is becoming a method of choice for studying gene function, but constructing conditional alleles is often laborious, limited by target gene structure, and at times, prone to incomplete conditional ablation. To address these issues, we developed a technology termed conditionals by inversion (COIN). Before activation, COINs contain an inverted module (COIN module) that lies inertly within the antisense strand of a resident gene. When inverted into the sense strand by a site-specific recombinase, the COIN module causes termination of the target gene's transcription and simultaneously provides a reporter for tracking this event. COIN modules can be inserted into natural introns (intronic COINs) or directly into coding exons as part of an artificial intron (exonic COINs), greatly simplifying allele design and increasing flexibility over previous conditional KO approaches. Detailed analysis of over 20 COIN alleles establishes the reliability of the method and its broad applicability to any gene, regardless of exon-intron structure. Our extensive testing provides rules that help ensure success of this approach and also explains why other currently available conditional approaches often fail to function optimally. Finally, the ability to split exons using the COIN's artificial intron opens up engineering modalities for the generation of multifunctional alleles.

SUBMITTER: Economides AN 

PROVIDER: S-EPMC3752204 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

altmetric image

Publications

Conditionals by inversion provide a universal method for the generation of conditional alleles.

Economides Aris N AN   Frendewey David D   Yang Peter P   Dominguez Melissa G MG   Dore Anthony T AT   Lobov Ivan B IB   Persaud Trikaldarshi T   Rojas Jose J   McClain Joyce J   Lengyel Peter P   Droguett Gustavo G   Chernomorsky Rostislav R   Stevens Sean S   Auerbach Wojtek W   Dechiara Thomas M TM   Pouyemirou William W   Cruz Joseph M JM   Feeley Kieran K   Mellis Ian A IA   Yasenchack Jason J   Hatsell Sarah J SJ   Xie Liqin L   Latres Esther E   Huang Lily L   Zhang Yuhong Y   Pefanis Evangelos E   Skokos Dimitris D   Deckelbaum Ron A RA   Croll Susan D SD   Davis Samuel S   Valenzuela David M DM   Gale Nicholas W NW   Murphy Andrew J AJ   Yancopoulos George D GD  

Proceedings of the National Academy of Sciences of the United States of America 20130805 34


Conditional mutagenesis is becoming a method of choice for studying gene function, but constructing conditional alleles is often laborious, limited by target gene structure, and at times, prone to incomplete conditional ablation. To address these issues, we developed a technology termed conditionals by inversion (COIN). Before activation, COINs contain an inverted module (COIN module) that lies inertly within the antisense strand of a resident gene. When inverted into the sense strand by a site-  ...[more]

Similar Datasets

| S-EPMC8092013 | biostudies-literature
| S-EPMC9954577 | biostudies-literature
| S-EPMC7849380 | biostudies-literature
| S-EPMC7089657 | biostudies-literature
| S-EPMC3435482 | biostudies-literature
| S-EPMC6845224 | biostudies-literature
| S-EPMC6709553 | biostudies-literature
| S-EPMC3961003 | biostudies-other
| S-EPMC30411 | biostudies-literature
| S-EPMC9943854 | biostudies-literature