Literature DB >> 18982302

Preparation of efficient splicing extracts from whole cells, nuclei, and cytoplasmic fractions.

Naoyuki Kataoka1, Gideon Dreyfuss.   

Abstract

Pre-mRNA (messenger RNA) splicing is an essential step for gene expression in higher eukaryotes. Splicing reactions have been well studied in vitro using extracts prepared from cultured cells. We describe protocols for the preparation of splicing-competent extracts from whole cells, nuclei, and cytoplasmic fractions. The nuclear and whole-cell extracts are fully active in splicing, while S100 extracts are able to support splicing only when SR (Serine/Arginine-rich) proteins are supplied. The simple method described here to prepare splicing active extracts from whole cells is particularly useful in studying pre-mRNA splicing in many different cell types.

Entities:  

Mesh：

Substances：

Year:  2008        PMID： 18982302     DOI： 10.1007/978-1-60327-475-3_23

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN： 1064-3745


  16 in total

1.  Contribution of DEAH-box protein DHX16 in human pre-mRNA splicing.

Authors:  Marieta Gencheva; Mitsuo Kato; Alain N S Newo; Ren-Jang Lin
Journal:  Biochem J       Date:  2010-07-01       Impact factor: 3.857

Review 2.  Assays for the identification and prioritization of drug candidates for spinal muscular atrophy.

Authors:  Jonathan J Cherry; Dione T Kobayashi; Maureen M Lynes; Nikolai N Naryshkin; Francesco Danilo Tiziano; Phillip G Zaworski; Lee L Rubin; Jill Jarecki
Journal:  Assay Drug Dev Technol       Date:  2014-08       Impact factor: 1.738

3.  A role for SMN exon 7 splicing in the selective vulnerability of motor neurons in spinal muscular atrophy.

Authors:  Matteo Ruggiu; Vicki L McGovern; Francesco Lotti; Luciano Saieva; Darrick K Li; Shingo Kariya; Umrao R Monani; Arthur H M Burghes; Livio Pellizzoni
Journal:  Mol Cell Biol       Date:  2011-10-28       Impact factor: 4.272

4.  Human spliceosomal protein CWC22 plays a role in coupling splicing to exon junction complex deposition and nonsense-mediated decay.

Authors:  Andrei Alexandrov; David Colognori; Mei-Di Shu; Joan A Steitz
Journal:  Proc Natl Acad Sci U S A       Date:  2012-12-10       Impact factor: 11.205

5.  C16orf57, a gene mutated in poikiloderma with neutropenia, encodes a putative phosphodiesterase responsible for the U6 snRNA 3' end modification.

Authors:  Seweryn Mroczek; Joanna Krwawicz; Jan Kutner; Michal Lazniewski; Iwo Kuciński; Krzysztof Ginalski; Andrzej Dziembowski
Journal:  Genes Dev       Date:  2012-08-16       Impact factor: 11.361

6.  A quantitative high-throughput in vitro splicing assay identifies inhibitors of spliceosome catalysis.

Authors:  Michael G Berg; Lili Wan; Ihab Younis; Michael D Diem; Michael Soo; Congli Wang; Gideon Dreyfuss
Journal:  Mol Cell Biol       Date:  2012-01-17       Impact factor: 4.272

7.  Preparation of splicing competent nuclear extracts.

Authors:  Chiu-Ho T Webb; Klemens J Hertel
Journal:  Methods Mol Biol       Date:  2014

8.  In vivo selection of kinase-responsive RNA elements controlling alternative splicing.

Authors:  Hongzhao Li; Guodong Liu; Jiankun Yu; Wenguang Cao; Vincent G Lobo; Jiuyong Xie
Journal:  J Biol Chem       Date:  2009-04-22       Impact factor: 5.157

9.  Preparation of Splicing Competent Nuclear Extract from Mammalian Cells and In Vitro Pre-mRNA Splicing Assay.

Authors:  Maliheh Movassat; Hossein Shenasa; Klemens J Hertel
Journal:  Methods Mol Biol       Date:  2017

10.  Genomics-guided discovery of thailanstatins A, B, and C As pre-mRNA splicing inhibitors and antiproliferative agents from Burkholderia thailandensis MSMB43.

Authors:  Xiangyang Liu; Sreya Biswas; Michael G Berg; Christopher M Antapli; Feng Xie; Qi Wang; Man-Cheng Tang; Gong-Li Tang; Lixin Zhang; Gideon Dreyfuss; Yi-Qiang Cheng
Journal:  J Nat Prod       Date:  2013-03-21       Impact factor: 4.050
View more

北京卡尤迪生物科技股份有限公司 © 2022-2023.