Protocol for identification and validation of IGF2BP1 target genes in pluripotent human embryonic carcinoma cells

We present a detailed protocol to identify and validate IGF2BP1 target genes in pluripotent human embryonic carcinoma cells (NTERA-2). We first identify the target genes through RNA-immunoprecipitation (RIP) sequencing. We then validate the identified targets through the use of RIP-qPCR assays, dete...

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Detalles Bibliográficos
Autores principales: Myint, Khine, Chuang, Linda Shyue Huey, Matsuo, Junichi, Ito, Yoshiaki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10227443/
https://www.ncbi.nlm.nih.gov/pubmed/37243602
http://dx.doi.org/10.1016/j.xpro.2023.102338
Descripción
Sumario:We present a detailed protocol to identify and validate IGF2BP1 target genes in pluripotent human embryonic carcinoma cells (NTERA-2). We first identify the target genes through RNA-immunoprecipitation (RIP) sequencing. We then validate the identified targets through the use of RIP-qPCR assays, determine the m(6)A status of target genes by m(6)A-IP, and perform functional validation by quantifying changes in mRNA or protein expression levels upon knockdown of IGF2BP1 or methyltransferases in NTERA-2. For complete details on the use and execution of this protocol, please refer to Myint et al. (2022).(1)

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