RNAseq data to study PRPF6 regulated splice forms in colon cancer cell lines
Alternative splicing plays critical roles in differentiation, development, and cancer (Pettigrew et al., 2008; Chen and Manley, 2009). The recent identification of specific spliceosome inhibitors has generated interest in the therapeutic potential of targeting this cellular process (van Alphen et al., 2009). Using an integrated genomic approach, we have identified PRPF6, an RNA binding component of the pre-mRNA spliceosome, as an essential driver of oncogenesis in colon cancer. Importantly, PRPF6 is both amplified and overexpressed in colon cancer, and only colon cancer cells with high PRPF6 levels are sensitive to its loss. Our data clearly point to an important role for PRPF6 in colon cancer growth and suggest that a better understanding of its role in alternative splicing in colon cancer is warranted. To determine the specific alternative splice forms that PRPF6 regulates in colon cancer, we plan three experiments: 1. The first involves knocking down expression of PRPF6 in two different cancer cell lines with 3 different siRNAs, and then completing RNA-seq to determine the gene expression changes that occur relative to a non-targeting control siRNA. Because of the role for PRPF6 in pre-mRNA splicing, we especially want to quantify the changes in splice-specific forms of all genes genome-wide to identify genes whose splicing is altered upon PRPF6 knockdown. 2. The second involves immunoprecipitating PRPF6 from two different cancer cell lines and isolating any RNA that is bound to PRPF6, since PRPF6 is an RNA-binding protein. We then want to carry out RNA-seq to identify which RNA molecules co-immunoprecipitated with PRPF6. This will help us determine possible functions for PRPF6 in regulating colon cancer growth. 3. The third involves overexpressing PRPF6 in cell lines and then carrying out RNA-seq to identify any changes in splice-specific gene expression. This will allow us to determine whether increased PRPF6 expression is sufficient to drive alternative splicing changes.
- 27/05/2015
- 34 samples
- DAC: EGAC00001000055
- Technology: Illumina HiSeq 2000
GENENTECH DATA ACCESS COMMITTEEDATA ACCESS AGREEMENT
GENENTECH DATA ACCESS COMMITTEEDATA ACCESS AGREEMENT WITH COMMERCIAL ORGANIZATIONThis Data Access Agreement (“Agreement”) between Genentech, Inc. (“Genentech”) and User and User Organization (as defined below) governs the terms on which access will be granted to the genotype data obtained from Genentech.In signing this Agreement, User and User Organization agree to be bound by the terms and conditions of access set out in this Agreement.For the sake of clarity, the terms of access set out in this Agreement apply both to the User and the User Organization (as defined below). User Organization and User are referred to within the Agreement as “You” and “Your” shall be construed accordingly.Definitions: gRED means Genentech Research and Early Development.Data means all and any human genetic data obtained from Genentech and generated by gRED. Data Subject means a person, who has been informed of the purpose for which the Data is held and has given his/her informed consent thereto.User means a researcher whose User Organization has previously completed this Data Access Agreement and has received acknowledgement of its acceptance.Publications means, without limitation, articles published in print journals, electronic journals, reviews, books, posters and other written and verbal presentations of research.User Organization means the commercial organization at which the User is employed or affiliated.Genentech Data Access Committee means members of the gRED organization that is responsible for making the data access decisions for the genotype data generated by gRED.Terms and Conditions: In signing this Agreement:1.You agree to use the Data only for the advancement of medical research.2.You agree to preserve, at all times, the confidentiality of Data pertaining to Data Subjects. In particular, You undertake not to use, or attempt to use the Data to compromise or otherwise infringe the confidentiality of information on Data Subjects and their right to privacy.3.You agree not to attempt to link the Data provided under this Agreement to other information or archive data available for the data sets provided, even if access to that data has been formally granted to you, or it is freely available without restriction, without specific permission being sought from the relevant access committees.4.You agree not to transfer or disclose the Data, in whole or part, or any identifiable material derived from the Data, to others, except as necessary for data/safety monitoring or program management. Should You wish to share the Data with a collaborator outside the same Organization, the third party must make a separate application for access to the Data.5.You agree to use the Data for the approved purpose and project described in your application; use of the Data for a new purpose or project will require a new application and approval.6.You shall be free to publish the results of the research using the Data for the approved purpose. You agree to acknowledge in any such publication that the research makes use of Data generated by Genentech/gRED.7.You accept that Genentech:a.bears no legal responsibility for the accuracy or comprehensiveness of the Data; and b.accepts no liability for indirect, consequential, or incidental, damages or losses arising from use of the Data, or from the unavailability of, or break in access to, the Data for whatever reason.8.You understand and acknowledge that the Data is protected by copyright and other intellectual property rights, and that duplication, except as reasonably required to carry out Your research with the Data, or sale of all or part of the Data on any media is not permitted.9.You recognize that nothing in this Agreement shall operate to transfer to the User Organization any intellectual property rights relating to the Data. The User Organization has the right to develop intellectual property based on the comparisons with their own data. Any invention or discovery made by User and/or User Organization arising out of the use of the Data (each a “Data Invention”) shall be promptly disclosed to Genentech in writing. User Organization hereby grants to Genentech a royalty-free, fully-paid, non-exclusive license (i) to make, use and import such Data Invention for internal research purposes; and (ii) to make, use, offer to sell, sell and import such Data Invention to the extent necessary to commercialize Genentech product(s), which license shall be sublicensable to Genentech’s affiliates and development and commercialization collaborators on such product(s). 10.You agree that you will submit a report to the Genentech Data Access Committee, if requested, on completion of the agreed purpose. The Genentech Data Access Committee agrees to treat the report and all information, data, results, and conclusions contained within such report as confidential information belonging to the User Organization.11.(a) You agree to use the Data in accordance with all applicable laws, including but not limited to applicable data privacy laws.(b) The Genentech Data Access Committee reserves the right to request and inspect data security and management documentation to ensure the adequacy of data protection measures. 12.This Agreement shall be effective as of the date of the last party to sign below (“Effective Date”) and shall terminate three (3) years after the Effective Date, unless terminated sooner pursuant to Section 13 below.13.You accept that this Agreement will terminate immediately upon any breach of this Agreement by You and You will be required to destroy any Data held. Either Party may terminate this Agreement upon thirty (30) days written notice for any reason. The following Sections of this Agreement shall survive its expiration or termination: Sections 1 – 10, 11(a), and 13. For and on behalf of User/Applicant:___________________________Signature of User/Applicant___________________________Print Name of User/Applicant___________________________Title___________________________DateFor and on behalf of User Organization:___________________________Signature of Organizational or Administrative Authority___________________________Print Name___________________________Title___________________________Name of User Organization___________________________DateFor Genentech, Inc.:___________________________Signature___________________________Print Name___________________________Title___________________________Date
Studies are experimental investigations of a particular phenomenon, e.g., case-control studies on a particular trait or cancer research projects reporting matching cancer normal genomes from patients.
| Study ID | Study Title | Study Type |
|---|---|---|
| EGAS00001000761 | Other |
This table displays only public information pertaining to the files in the dataset. If you wish to access this dataset, please submit a request. If you already have access to these data files, please consult the download documentation.