| Chapter title |
Comprehensive Identification of mRNA Polyadenylation Sites by PAPERCLIP
|
|---|---|
| Chapter number | 7 |
| Book title |
mRNA Processing
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-7204-3_7 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7203-6, 978-1-4939-7204-3
|
| Authors |
Hun-Way Hwang, Robert B. Darnell |
| Abstract |
We discuss a newly developed method to profile mRNA polyadenylation (pA) sites in an unbiased manner, PAPERCLIP (Poly(A) binding Protein-mediated mRNA 3'End Retrieval by CrossLinking ImmunoPrecipitation). Based on the well-established CLIP (crosslinking immunoprecipitation) technique, PAPERCLIP utilizes the poly(A) binding protein (PABP) as a biological filter to selectively retrieve mRNA 3' end fragments by immunoprecipitation from ultraviolet (UV) irradiated tissues or cultured cells. The mRNA fragments are subsequently extracted from the immunoprecipitated PABP:RNA complexes to generate a cDNA library, which goes through two rounds of purification before the final amplification by real-time polymerase chain reaction (PCR). The amplified cDNA library can then be read out by high-throughput sequencing to generate a transcriptomic profile and comprehensive alternative poly(A) (APA) site map from intact tissue or cultured cells. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 6 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 1 | 17% |
| Professor > Associate Professor | 1 | 17% |
| Student > Doctoral Student | 1 | 17% |
| Unknown | 3 | 50% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 50% |
| Unknown | 3 | 50% |