gRNA subjected to wild type T cell TCR alpha chain knockout and method

Abstract

The invention discloses a gRNA subjected to wild type T cell TCR alpha chain knockout and a method. The sequence of the gRNA is shown as SEQ ID NO:1. By using a CRISPR / Cas 9 technology, the gRNA and the CRISPR / Cas9 perform co-infection on T cells; the wild type T cell TCR alpha chain knockout is performed; the T cells lack of wild type T cell TCR alpha chains are built; the gRNA can be used for CAR-T or TCR-T cellular immunity treatment. The gRNA has high knockout rate; the preparation method is relatively simple and easy; the T cells lack of wild type T cell TCR alpha chains can be fast and efficiently provided for clinics.

Description

technical field [0001] The invention relates to the technical field of gene knockout, in particular to a gRNA and method for knocking out TCR alpha chain of wild-type T cells. Background technique [0002] Tumor is currently the biggest killer of human health. Existing treatments including surgery, chemotherapy, and radiotherapy cannot completely eradicate tumors. Immune T cell therapy has brought great hope for the recovery of tumor patients. At present, the methods of immune T cell therapy mainly include CAR-T and TCR-T cell technology. The principle of these T cell therapy is to transfect tumor-specific chimeric antigen receptor (CAR) or T cell receptor (TCR) gene into normal T cells. Above, make it acquire the ability of tumor-specific recognition, so as to kill tumor cells. However, since normal T cells contain their own T cell receptors (wild-type alpha and beta chains), these wild-type alpha and beta chains can affect the expression of CAR-T or TCR-T, resulting in CA...

AI Technical Summary

Problems solved by technology

However, the above technologies also have many shortcomings in practical applications. Among them, RNAi technology only reduces the expression of wild-type alpha and beta chains at the RNA level, and it is an incomplete knockout, and the remaining RNA can continue to express TCR protein molecules ; ZFN technology requires the preparation of multiple vectors, which can be knocked out only after transcribed into RNA in vitro and then transferred into T cells. The RNA is unstable and cumbersome to prepare; Low, affecting the knockout efficiency of TCR wild-type alpha and beta chains

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