Establishment and characterization of mouse lines useful for endogenous protein degradation via an improved auxin‐inducible degron system (AID2) DOI Creative Commons

Hatsune Makino‐Itou,

Noriko Yamatani,

Akemi Okubo

и другие.

Development Growth & Differentiation, Год журнала: 2024, Номер 66(7), С. 384 - 393

Опубликована: Сен. 1, 2024

Abstract The development of new technologies opens avenues in the research field. Gene knockout is a key method for analyzing gene function mice. Currently, conditional strategies are employed to examine temporal and spatial function. However, phenotypes sometimes not observed because time required depletion due long half‐life target proteins. Protein knockdown using an improved auxin‐inducible degron system, AID2, overcomes such difficulties owing rapid efficient depletion. We AID‐tagged proteins within few several hours by simple intraperitoneal injection auxin analog, 5‐Ph‐IAA, which much shorter than knockout. Importantly, loss protein reversible, making useful measure effects transient Here, we also established mouse lines AID2‐medicated knockdown, include knock‐in ROSA26 locus; one expresses TIR1(F74G), other reporter expressing AID‐mCherry. germ‐cell‐specific TIR1 line confirmed specificity. In addition, introduced AID tag endogenous protein, DCP2 via CAS9‐mediated editing method. that was effectively eliminated resulted similar phenotype 20 h.

Язык: Английский

Establishment and characterization of mouse lines useful for endogenous protein degradation via an improved auxin‐inducible degron system (AID2) DOI Creative Commons

Hatsune Makino‐Itou,

Noriko Yamatani,

Akemi Okubo

и другие.

Development Growth & Differentiation, Год журнала: 2024, Номер 66(7), С. 384 - 393

Опубликована: Сен. 1, 2024

Abstract The development of new technologies opens avenues in the research field. Gene knockout is a key method for analyzing gene function mice. Currently, conditional strategies are employed to examine temporal and spatial function. However, phenotypes sometimes not observed because time required depletion due long half‐life target proteins. Protein knockdown using an improved auxin‐inducible degron system, AID2, overcomes such difficulties owing rapid efficient depletion. We AID‐tagged proteins within few several hours by simple intraperitoneal injection auxin analog, 5‐Ph‐IAA, which much shorter than knockout. Importantly, loss protein reversible, making useful measure effects transient Here, we also established mouse lines AID2‐medicated knockdown, include knock‐in ROSA26 locus; one expresses TIR1(F74G), other reporter expressing AID‐mCherry. germ‐cell‐specific TIR1 line confirmed specificity. In addition, introduced AID tag endogenous protein, DCP2 via CAS9‐mediated editing method. that was effectively eliminated resulted similar phenotype 20 h.

Язык: Английский

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