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Literature DB >> 24549659

Preparation of splicing competent nuclear extracts.

Abstract

Splicing components play an essential role in mediating accurate and efficient splicing. The complexity of the spliceosome and its regulatory networks increase the difficulty of studying the splicing reaction in detail. Nuclear extracts derived from HeLa cells provide all of the obligatory components to carry out intron removal in vitro. This chapter describes the large-scale preparation of nuclear extract from HeLa cells.

Entities: CellLine Chemical Disease Gene Species

Mesh：

Substances：
Cell Extracts

Year: 2014 PMID： 24549659 PMCID： PMC4501576 DOI： 10.1007/978-1-62703-980-2_8

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

14 in total

1. Preparation of HeLa cell nuclear and cytosolic S100 extracts for in vitro splicing.

Authors: A Mayeda; A R Krainer
Journal: Methods Mol Biol Date: 1999

2. A simple whole cell lysate system for in vitro splicing reveals a stepwise assembly of the exon-exon junction complex.

Authors: Naoyuki Kataoka; Gideon Dreyfuss
Journal: J Biol Chem Date: 2003-11-18 Impact factor: 5.157

Preparation of splicing competent nuclear extracts.

1. Preparation of HeLa cell nuclear and cytosolic S100 extracts for in vitro splicing.

2. A simple whole cell lysate system for in vitro splicing reveals a stepwise assembly of the exon-exon junction complex.

3. Analyzing mechanisms of alternative pre-mRNA splicing using in vitro splicing assays.

4. Preparation of nuclear and cytoplasmic extracts from mammalian cells.

5. Purification and characterization of pre-mRNA splicing factor SF2 from HeLa cells.

6. Transcription and processing of intervening sequences in yeast tRNA genes.

7. Preparation of HeLa nuclear extracts.

8. Normal and mutant human beta-globin pre-mRNAs are faithfully and efficiently spliced in vitro.

9. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.

10. PARP1 represses PAP and inhibits polyadenylation during heat shock.

1. RNA-binding proteins with basic-acidic dipeptide (BAD) domains self-assemble and aggregate in Alzheimer's disease.

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