TY - JOUR
T1 - Highly efficient transgenesis with miniMos in Caenorhabditis briggsae
AU - Ding, Qiutao
AU - Ren, Xiaoliang
AU - Li, Runsheng
AU - Chan, Luyan
AU - Ho, Vincy Wing Sze
AU - Bi, Yu
AU - Xie, Dongying
AU - Zhao, Zhongying
N1 - Funding Information:
We thank Ms Cindy Tan for logistic support and members of Zhao’s lab for helpful comments. Some strains were provided by the CGC, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440).
Funding Information:
This work was supported by General Research Funds (N_HKBU201/18, HKBU12100118, HKBU12101520, HKBU12101522) from Hong Kong Research Grant Council, and Initiation Grant for Faculty Niche Research Areas RC-FNRA-IG/21-22/SCI/02 from Hong Kong Baptist University to ZZ.
Publisher Copyright:
© 2022 Genetics Society of America. All rights reserved.
PY - 2022/12
Y1 - 2022/12
N2 - Caenorhabditis briggsae as a companion species for Caenorhabditis elegans has played an increasingly important role in study of evolution of development and genome and gene regulation. Aided by the isolation of its sister spices, it has recently been established as a model for speciation study. To take full advantage of the species for comparative study, an effective transgenesis method especially those with single-copy insertion is important for functional comparison. Here, we improved a transposon-based transgenesis methodology that had been originally developed in C. elegans but worked marginally in C. briggsae. By incorporation of a heat shock step, the transgenesis efficiency in C. briggsae with a single-copy insertion is comparable to that in C. elegans. We used the method to generate 54 independent insertions mostly consisting of a mCherry tag over the C. briggsae genome. We demonstrated the use of the tags in identifying interacting loci responsible for hybrid male sterility between C. briggsae and Caenorhabditis nigoni when combined with the GFP tags we generated previously. Finally, we demonstrated that C. briggsae tolerates the C. elegans toxin, PEEL-1, but not SUP-35, making the latter a potential negative selection marker against extrachromosomal array.
AB - Caenorhabditis briggsae as a companion species for Caenorhabditis elegans has played an increasingly important role in study of evolution of development and genome and gene regulation. Aided by the isolation of its sister spices, it has recently been established as a model for speciation study. To take full advantage of the species for comparative study, an effective transgenesis method especially those with single-copy insertion is important for functional comparison. Here, we improved a transposon-based transgenesis methodology that had been originally developed in C. elegans but worked marginally in C. briggsae. By incorporation of a heat shock step, the transgenesis efficiency in C. briggsae with a single-copy insertion is comparable to that in C. elegans. We used the method to generate 54 independent insertions mostly consisting of a mCherry tag over the C. briggsae genome. We demonstrated the use of the tags in identifying interacting loci responsible for hybrid male sterility between C. briggsae and Caenorhabditis nigoni when combined with the GFP tags we generated previously. Finally, we demonstrated that C. briggsae tolerates the C. elegans toxin, PEEL-1, but not SUP-35, making the latter a potential negative selection marker against extrachromosomal array.
KW - Caenorhabditis briggsae
KW - heat shock
KW - miniMos
KW - transgenesis
UR - http://dx.doi.org/10.1093/g3journal/jkac254
U2 - 10.1093/g3journal/jkac254
DO - 10.1093/g3journal/jkac254
M3 - Journal article
SN - 2160-1836
VL - 12
JO - G3: Genes, Genomes, Genetics
JF - G3: Genes, Genomes, Genetics
IS - 12
M1 - jkac254
ER -