Nuclear Pre-mRNA Splicing in Vitro PDF Download

Author: Adrian Robert Krainer
Publisher:
ISBN:
Category : Genetic transcription
Languages : en
Pages : 462

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Author: Klemens J. Hertel
Publisher: Humana
ISBN: 9781627039796
Category : Medical
Languages : en
Pages : 0

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Book Description
Providing a guide to classical experimental approaches to decipher splicing mechanisms and experimental strategies that rely on novel multi-disciplinary approaches, Spliceosomal Pre-mRNA Splicing: Methods and Protocols describes the theory of alternative pre-mRNA splicing in seven introductory chapters and then introduces protocols and their theoretical background relevant for a variety of experimental research. These protocol chapters cover basic methods to detect splicing events, analyses of alternative pre-mRNA splicing in vitro and in vivo manipulation of splicing events and high-throughput and bioinformatic analyses of alternative splicing. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Spliceosomal Pre-mRNA Splicing: Methods and Protocols will aid newcomers and seasoned molecular biologists in understanding the fascinating world of alternative splicing with the ultimate goal of paving the way for many new discoveries to come.

Author: Stefan Stamm
Publisher: John Wiley & Sons
ISBN: 3527326065
Category : Science
Languages : en
Pages : 660

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Book Description
Der definitive Leitfaden zum RNA-Splicing - ideal für alle Kliniker, die sich mit genetischen Erkrankungen befassen, insbesondere natürlich für RNA-Forscher. Website: www.wiley-vch.de/home/splicing

Author: A. S. N. Reddy
Publisher: Springer Science & Business Media
ISBN: 3540767762
Category : Science
Languages : en
Pages : 323

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Book Description
During the last few years, tremendous progress has been made in understanding various aspects of pre-mRNA processing. This book, with contributions from leading scientists in this area, summarizes recent advances in nuclear pre-mRNA processing in plants. It provides researchers in the field, as well as those in related areas, with an up-to-date and comprehensive, yet concise, overview of the current status and future potential of this research in understanding plant biology.

Author: Evelyn Jane Strauss
Publisher:
ISBN:
Category : Genetic engineering
Languages : en
Pages : 340

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Author: Angus I. Lamond
Publisher: Springer Science & Business Media
ISBN: 3662223252
Category : Science
Languages : en
Pages : 230

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he past fifteen years have seen tremendous growth in our understanding of T the many post-transcriptional processing steps involved in producing func tional eukaryotic mRNA from primary gene transcripts (pre-mRNA). New processing reactions, such as splicing and RNA editing, have been discovered and detailed biochemical and genetic studies continue to yield important new insights into the reaction mechanisms and molecular interactions involved. It is now apparent that regulation of RNA processing plays a significant role in the control of gene expression and development. An increased understanding of RNA processing mechanisms has also proved to be of considerable clinical importance in the pathology of inherited disease and viral infection. This volume seeks to review the rapid progress being made in the study of how mRNA precursors are processed into mRNA and to convey the broad scope of the RNA field and its relevance to other areas of cell biology and medicine. Since one of the major themes of RNA processing is the recognition of specific RNA sequences and structures by protein factors, we begin with reviews of RNA-protein interactions. In chapter 1 David Lilley presents an overview of RNA structure and illustrates how the structural features of RNA molecules are exploited for specific recognition by protein, while in chapter 2 Maurice Swanson discusses the structure and function of the large family of hnRNP proteins that bind to pre-mRNA. The next four chapters focus on pre-mRNA splicing.

Author: Shin-Lian Doong
Publisher:
ISBN:
Category :
Languages : en
Pages : 254

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Author: Neha Aggarwal
Publisher:
ISBN:
Category : RNA splicing
Languages : en
Pages : 75

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Nuclear pre-mRNA splicing is an important step in eukaryotic RNA processing and can yield a variety of transcripts. Further, it is known that nuclear pre-mRNA introns are of two types, U2-dependent and U12-dependent. Recently discovered, microRNAs (miRNA) are short, endogenous, 20-22 nucleotide long, non-coding RNA molecules that base pair with target mRNAs to modulate translation. Current evidence suggests that processing of intronic miRNA does not affect the splicing. Since, intronic miRNAs are involved in essential cellular processes, we surmised if splice sites of miRNA coding introns are flexible to support productive precursor and mature miRNA processing. We constructed a minigene reporter consisting exons 26-29 including introns from MYH6 gene and an intronic miRNA, miR-208 harbored in intron 27. Using in vivo and in vitro methods, we examined splicing of MYH6 intron 27 and processing of miR-208. The predicted stem-loop structure of miR-208 and the 5' splice site of the intron 27 were mutated. Stem-loop mutations had no recordable splicing defect of intron 27. The 5'SS mutant activated a cryptic splicing event using a sub-optimal splice site located 50 nucleotides upstream of wild-type splice site. Interestingly, conversion of U2-type 5' splice site to consensus U12-type 5' splice site did not affect in vivo splicing of the intron, suggesting potential conversion of intron to U12-type, albeit without a consensus U12-type branch site sequence. However, miRNA processing was affected as established by in vitro splicing assay and qRT-PCR analysis of miR-208. We mutated nucleotides from position 3 to 7 in intron 27 to study if changes in splice site will allow miRNA processing to occur and if splicing will also be affected. Our data indicates significant changes in spliced phenotype as well as in miR-208 expression level. Since, all known intronic miRNAs are harbored in U2-dependent introns of mammalian genes, we wanted to study if miRNA processing will be compatible with U12-dependent intron. With this aim, we incorporated miR-208 in a U12-dependent intron. However, it appears that spliceosome and microprocessor cannot act together in a U12-dependent intron. This is first time that a miRNA has been incorporated in a U12-dependent intron to study its processing and also if splice sites are plastic for miRNA processing.

Author: Zairong Li
Publisher:
ISBN:
Category :
Languages : en
Pages : 370

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Author: Katharine Marie Lang
Publisher:
ISBN:
Category :
Languages : en
Pages : 308

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