Cas9 RNP targeting cis-regulatory elements in lymphocytes

Cas9 RNP 靶向淋巴细胞中的顺式调控元件

• 批准号: 10630567
• 负责人: Karl Mark Ansel
• 金额: $ 7.84万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10630567
• 关键词: Achievement; Animal Model; Asthma; B-Lymphocytes; Base Pairing; Binding; Biochemical; Bioinformatics; Biological; CRISPR library; CRISPR/Cas technology; Cell Culture Techniques; Cell physiology; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Complex; Development; Doctor of Philosophy; Family; Gene Expression; Genes; Genetic; Genomics; Goals; Guide RNA; Human; Hypersensitivity; Immune; Immune response; Individual; Investigation; Laboratories; Libraries; Lymphocyte; Lymphocyte Function; Mediating; Messenger RNA; MicroRNAs; Modeling; Molecular; Mus; Nucleic Acid Regulatory Sequences; Open Reading Frames; Pathogenesis; Pathway interactions; Play; Proteins; Regulation; Regulator Genes; Regulatory Element; Research; Research Personnel; Role; Somatic Cell; Specialist; Specificity; System; T-Lymphocyte; Technical Expertise; Testing; Transfection; Translations; Work; base; career; design; experience; experimental study; gene discovery; gene network; genome editing; immunopathology; mRNA Transcript Degradation; next generation; novel; parent grant; prevent; programs; skills; success; tool

Project Summary CRISPR tools for genome editing have revolutionized and accelerated the functional interrogation of coding genes in model animal systems and in human somatic cells, including primary lymphocytes that play critical roles in the immunopathology of allergy and asthma. We propose to adapt and optimize these tools for miRNA- directed pathway discovery. First, will create pooled retroviral CRISPR libraries for targets that we have empirically identified for miR-29, a miRNA with critical functions in both T cells and B cells. Second, we will extend next generation transfection-based CRISPR systems to target cis-regulatory sequences that do not code for protein, but mediate regulatory interactions between miRNAs and target mRNAs. Success in these aims will enable detailed mechanistic analysis of miRNA:target networks that govern immune responses in asthma and allergy. A second important goal of this proposal is to support the career and scientific development of Celeste Garza, a post-baccalaureate researcher gaining research experience as a junior specialist in the Ansel laboratory. The proposed experiments build on skills she has learned in the Ansel lab, and will provide the opportunity for her to develop a research narrative and scientific achievements as prepares to apply for PhD programs in the coming year.

项目总结