Long undecoded transcript isoform (LUTI) detection in meiotic budding yeast by direct RNA and transcript leader sequencing

Summary: LUTIs (Long Undecoded Transcript Isoforms) are 5′-extended and poorly translated mRNAs that can downregulate transcription from promoters more proximal to a gene’s coding sequence (CDS). In this protocol, polyA RNA is extracted from budding yeast cells undergoing highly synchronized meiosis...

وصف كامل

التفاصيل البيبلوغرافية
المؤلفون الرئيسيون: Amy Tresenrider, Minghao Chia, Folkert J. van Werven, Elçin Ünal
التنسيق: مقال
اللغة:English
منشور في: Elsevier 2022-03-01
سلاسل:STAR Protocols
الموضوعات:
الوصول للمادة أونلاين:http://www.sciencedirect.com/science/article/pii/S2666166722000259
الوصف
الملخص:Summary: LUTIs (Long Undecoded Transcript Isoforms) are 5′-extended and poorly translated mRNAs that can downregulate transcription from promoters more proximal to a gene’s coding sequence (CDS). In this protocol, polyA RNA is extracted from budding yeast cells undergoing highly synchronized meiosis. Using a combination of long-read direct RNA sequencing and transcript leader sequencing (TL-seq), meiosis-specific LUTIs are systematically identified. Following identification, TL-seq is used to quantify the abundance of both LUTI and the more canonical gene-proximal (PROX) transcripts.For complete details on the use and execution of this protocol, please refer to Tresenrider et al. (2021).
تدمد:2666-1667