DESCRIPTION: See instructions. State the application's broad, long-term objectives and specific aims, making referenceto the health relatedness of the project (i.e., relevance to the mission of the agency). Describe concisely the research design and methods for achieving these goals. Describe the rationale and techniques you will use to pursue these goals. In addition, in two or three sentences, describe in plain, lay language the relevance of this research to public health. If the application is funded, this description, as is, will become public information. Therefore, do not include proprietary/confidential information. DO NOT EXCEED THE SPACE PROVIDED. This is an application for a supplement to a K18 grant. The purpose of the supplement is to provide funds to permit initiation of embryonic stem cell research in my laboratory to replace the reagents generated during my K18 grant-supported research Waningin Australia, which have become unavailable for transfer to my aboratory due to unforeseen circumstances beyond the control of my research mentor (Dr. Martin Pera), my collaborator in the Pera lab (Dr. Ernst Wolvetang), or me. With this supplemental support, my lab will be able to purchase 3 human US federally approved human embryonic stem cell (hESC) lines, lentiviral expression vectors to transduce the embryonic stem cells, and oligonucleotides (plus sequencing confirmation) for construction of expression vectors. These funds will allow us to extablish growth of hESC lines and to transduce these hESC lines with lentiviral expression vectors. Once we have completed these steps, we will have reconstructed what we created in Australia and will be prepared to initiate our own research program on regulation of hESC sternness and differentiation. PERFORMANCE SITE(S) (organization, city, state) H. Lee Moffitt Cancer Center and Research Institute University of South Florida Tampa, FL PHS 398 (Rev. 04/06) Page 2 Form Page 2 Principal Investigator/Program Director (Last, First, Middle): ZllCkerman, Kenneth S. KEY PERSONNEL. See instructions. Use continuation pages as neededto provide the required information in the formatshown below. Start with Principal Investigator(s).List all other key personnel in alphabetical order, last name first. Name eRA Commons User Name Organization Role on Project Kenneth S. Zuckerman kenzuckerman H. Lee Moffitt Cancer CtrP.I. OTHER SIGNIFICANT CONTRIBUTORS Name Organization Role on Project Human Embryonic Stem Cells Q No ^ Yes If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://StemcellS.nih.qov/registrv/index.asp. Usecontinuation pages as needed. If a specific line cannot be referenced at this time,include a statementthat one from the Registry will be used. Cell Line WA01 WA07 WA09 PHS 398 (Rev. 04/06) Page 3 Form Page 2-continued Number the following pages consecutively throughout the application. Do not use suffixes such as 4a, 4b. Principal Investigator/Program Director (Last, First, Middle): ZllCkerman, Kenneth S. The name of the principal investigator/program director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT TABLE OFCONTENTS Page Numbers Face Page 1 Description, Performance Sites, Key Personnel, Other Significant Contributors, and Human Embryonic Stem Cells 2-3