Genetic Manipulation of SAM Synthetases in C. elegans

N2(Caenorhabditis Genetics Center); sams-1(lof)(ok3033); sams-3(ok2932) IV, sams-4((ok3315) IV, Caenorhabditis Genetics Center), tagRFP::SAMS-1 (WAL500, this study); GFP::SAM-4(WALK501, this study); SAMS-1::RFP;GFP::SAMS-4(WAL502, this study), SAMS-3::mKate (WAL305). Pges-1::NHR-68::GFP (VL1296) (Bulcha et al., 2019 (link)). CRISPR tagging for WAL500 and WAL501 were done by the UMASS Medical School transgenic core, confirmed by PCR for genotype and outcrossed three times to wild type animals. Next, each strain was crossed to the respective deletion allele to create WAL503 (RFP::sams-1(ker5); sams-1(ok3033)) and WAL504(GFP::sams-4(ker6); sams-4(ok3315)). sams-3::mKate(nu3139) (COP2476) was constructed using CRISPR by In Vivo biosystems then outcrossed three times (WAL305).

Free full text: Click here

Godbole A.A., Gopalan S., Nguyen T.K., Munden A.L., Lui D.S., Fanelli M.J., Vo P., Lewis C.A., Spinelli J.B., Fazzio T.G, & Walker A.K. (2023). S-adenosylmethionine synthases specify distinct H3K4me3 populations and gene expression patterns during heat stress. eLife, 12, e79511.

Publication 2023

Allele Caenorhabditis Crispr Deletion Genotype Mkate Pges 1 Sams Strain Transgenic Wild animals

Corresponding Organization :

Other organizations : UMass Memorial Medical Center

Top 5 similar protocols

Variable analysis

independent variables

CRISPR tagging for WAL500 and WAL501
Crossing of each strain to the respective deletion allele to create WAL503 and WAL504
Construction of sams-3::mKate(nu3139) (COP2476) using CRISPR by In Vivo biosystems

dependent variables

TagRFP::SAMS-1 (WAL500, this study)
GFP::SAM-4 (WALK501, this study)
SAMS-1::RFP;GFP::SAMS-4 (WAL502, this study)
SAMS-3::mKate (WAL305)
Pges-1::NHR-68::GFP (VL1296)

control variables

Outcrossing of WAL500 and WAL501 strains three times to wild type animals
Outcrossing of COP2476 strain three times

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!