CRISPR/Cas9 Mediated High Efficiency Knockout of Myosin Essential Light Chain Gene in the Pacific Oyster (Crassostrea Gigas)

2021 ◽  
Author(s):  
Huijuan Li ◽  
Hong Yu ◽  
Shaojun Du ◽  
Qi Li
Keyword(s):  
Light Chain ◽  
Crassostrea Gigas ◽  
High Efficiency ◽  
Pacific Oyster ◽  
Chain Gene ◽  
Light Chain Gene ◽  
Essential Light Chain ◽  
The Pacific

Targeted disruption of the Dictyostelium myosin essential light chain gene produces cells defective in cytokinesis and morphogenesis

1995 ◽  
Vol 108 (10) ◽  
pp. 3207-3218 ◽  
Author(s):  
T.L. Chen ◽  
P.A. Kowalczyk ◽  
G. Ho ◽  
R.L. Chisholm
Keyword(s):  
Atpase Activity ◽  
Cell Line ◽  
Light Chain ◽  
Contractile Function ◽  
Cell Types ◽  
Chain Gene ◽  
Dictyostelium Cell ◽  
Wild Type ◽  
Light Chain Gene ◽  
Essential Light Chain

We have previously demonstrated that the myosin essential light chain (ELC) is required for myosin function in a Dictyostelium cell line, 7–11, in which the expression of ELC was inhibited by antisense RNA overexpression. We have now disrupted the gene encoding the ELC (mlcE) in Dictyostelium by gene targeting. The mlcE- mutants provide a clean genetic background for phenotypic analysis and biochemical characterization by removing complications arising from the residual ELC present in 7–11 cells, as well as the possibility of mutations due to insertion of the antisense construct at multiple sites in the genome. The mlcE- mutants, when grown in suspension, exhibited the typical multinucleate phenotype observed in both myosin heavy chain mutants and 7–11 cells. This phenotype was rescued by introducing a construct that expressed the wild-type Dictyostelium ELC cDNA. Myosin purified from the mlcE- cells exhibited significant calcium ATPase activity, but the actin-activated ATPase activity was greatly reduced. The results obtained from the mlcE- mutants strengthen our previous conclusion based on the antisense cell line 7–11 that ELC is critical for myosin function. The proper localization of myosin in mlcE- cells suggests that its phenotypic defects primarily arise from defective contractile function of myosin rather than its mislocalization. The enzymatic defect of myosin in mlcE- cells also suggests a possible mechanism for the observed chemotactic defect of mlcE- cells. We have shown that while mlcE- cells were able to respond to chemoattractant with proper directionality, their rate of movement was reduced. During chemotaxis, proper directionality toward chemoattractant may depend primarily on proper localization of myosin, while efficient motility requires contractile function. In addition, we have analyzed the morphogenetic events during the development of mlcE- cells using lacZ reporter constructs expressed from cell type specific promoters. By analyzing the morphogenetic patterns of the two major cell types arising during Dictyostelium development, prespore and prestalk cells, we have shown that the localization of prespore cells is more susceptible to the loss of ELC than prestalk cells, although localization of both cell types is abnormal when developed in chimeras formed by mixing equal numbers of wild-type and mutant cells. These results suggest that the morphogenetic events during Dictyostelium development have different requirements for myosin.

scholarly journals Structure and sequence of the myosin alkali light chain gene expressed in adult cardiac atria and fetal striated muscle.

1988 ◽  
Vol 263 (25) ◽  
pp. 12669-12676 ◽  
Author(s):  
P J Barton ◽  
B Robert ◽  
A Cohen ◽  
I Garner ◽  
D Sassoon ◽  
...  
Keyword(s):  
Light Chain ◽  
Striated Muscle ◽  
Chain Gene ◽  
Light Chain Gene

Combined effects of pollutants and salinity on embryo-larval development of the Pacific oyster, Crassostrea gigas

2016 ◽  
Vol 113 ◽  
pp. 31-38 ◽  
Author(s):  
Perrine Gamain ◽  
Patrice Gonzalez ◽  
Jérôme Cachot ◽  
Patrick Pardon ◽  
Nathalie Tapie ◽  
...  
Keyword(s):  
Larval Development ◽  
Crassostrea Gigas ◽  
Pacific Oyster ◽  
Combined Effects ◽  
The Pacific

Differentially methylated gene regions between resistant and susceptible heat-phenotypes of the Pacific oyster Crassostrea gigas

Aquaculture ◽  
2021 ◽  
pp. 736923
Author(s):  
Roberto Arredondo-Espinoza ◽  
Ana M. Ibarra ◽  
Steven B. Roberts ◽  
Maria Teresa Sicard-Gonzalez ◽  
Cristina Escobedo-Fregoso
Keyword(s):  
Crassostrea Gigas ◽  
Pacific Oyster ◽  
The Pacific ◽  
Differentially Methylated Gene
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