Analysis of the phytochrome gene family in Ceratodon purpureus by gene targeting reveals the primary phytochrome responsible for photo- and polarotropism

ABSTRACT Diva (also called Boo/Bcl-B) is a member of the Bcl-2 gene family and most likely functions during apoptosis. Diva is highly expressed in the ovary, and both pro- and antiapoptotic functions have been ascribed to this protein. To determine the role of Diva during murine development, we used gene targeting to inactivate Diva. The Diva-null mice are born at the expected ratios, are fertile, and have no obvious histological abnormalities, and long-term survival did not differ from littermate controls. Additionally, Diva was not required for apoptosis occurring from genotoxic insult in the ovaries or other organs. Thus, Diva is not critical for the normal development of the ovaries, or in its absence its function is subserved by another protein.

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Sequence similarities of phytochrome to protein kinases: implication for the structure, function and evolution of the phytochrome gene family

FEBS Letters ◽

10.1016/0014-5793(94)01327-w ◽

1995 ◽

Vol 357 (2) ◽

pp. 149-155 ◽

Cited By ~ 14

Author(s):

Fritz Thümmler ◽

Patricia Algarra ◽

Gisela M. Fobo

Keyword(s):

Structure Function ◽

Gene Family ◽

Protein Kinases ◽

Phytochrome Gene ◽

Sequence Similarities

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Multiplex CRISPR-Cas9 mutagenesis of the phytochrome gene family in Physcomitrium (Physcomitrella) patens

Plant Molecular Biology ◽

10.1007/s11103-020-01103-x ◽

2020 ◽

Author(s):

Silvia Trogu ◽

Anna Lena Ermert ◽

Fabian Stahl ◽

Fabien Nogué ◽

Tanja Gans ◽

...

Keyword(s):

Gene Family ◽

Lower Order ◽

Physcomitrella Patens ◽

Gene Families ◽

Mutant Library ◽

Highly Efficient ◽

Phytochrome Gene ◽

Efficient Screening ◽

Mutant Collection ◽

Proof Of Principle

Key message We mutated all seven Physcomitrium (Physcomitrella) patens phytochrome genes using highly-efficient CRISPR-Cas9 procedures. We thereby identified phy5a as the phytochrome primarily responsible for inhibiting gravitropism, proving the utility of the mutant library. Abstract The CRISPR-Cas9 system is a powerful tool for genome editing. Here we report highly-efficient multiplex CRISPR-Cas9 editing of the seven-member phytochrome gene family in the model bryophyte Physcomitrium (Physcomitrella) patens. Based on the co-delivery of an improved Cas9 plasmid with multiple sgRNA plasmids and an efficient screening procedure to identify high-order multiple mutants prior to sequencing, we demonstrate successful targeting of all seven PHY genes in a single transfection. We investigated further aspects of the CRISPR methodology in Physcomitrella, including the significance of spacing between paired sgRNA targets and the efficacy of NHEJ and HDR in repairing the chromosome when excising a complete locus. As proof-of-principle, we show that the septuple phy− mutant remains gravitropic in light, in line with expectations, and on the basis of data from lower order multiplex knockouts conclude that phy5a is the principal phytochrome responsible for inhibiting gravitropism in light. We expect, therefore, that this mutant collection will be valuable for further studies of phytochrome function and that the methods we describe will allow similar approaches to revealing specific functions in other gene families.

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Scnn1Sodium Channel Gene Family in Genetically Engineered Mice

Journal of the American Society of Nephrology ◽

10.1681/asn.v11suppl_2s129 ◽

2000 ◽

Vol 11 (suppl 2) ◽

pp. S129-S134 ◽

Cited By ~ 1

Author(s):

EDITH HUMMLER ◽

FRIEDRICH BEERMANN

Keyword(s):

Gene Family ◽

Gene Targeting ◽

Genetically Engineered ◽

Channel Gene ◽

Limiting Step ◽

Organ Systems ◽

Genetically Engineered Mice ◽

Perinatal Lethality ◽

Targeting Strategy ◽

A Current

Abstract.The amiloride-sensitive epithelial sodium channel is the limiting step in salt absorption. In mice, this channel is composed of three subunits (α, β, and γ), which are encoded by different genes (Scnn1a, Scnn1b, andScnn1c, respectively). The functions of these genes were recently investigated in transgenic (knockout) experiments, and the absence of any subunit led to perinatal lethality. More defined phenotypes have been obtained by introducing specific mutations or using transgenic rescue experiments. In this report, these approaches are summarized and a current gene-targeting strategy to obtain conditional inactivation of the channel is illustrated. This latter approach will be indispensable for the investigation of channel function in a wide variety of organ systems.

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