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US 2019/0376091 A1Published application
Primary cell gene editing
AssigneePact Pharma, Inc.
InventorsKyle Jacoby, Alex Franzusoff
Life of this patent
Estimating the term…
Abstract
Methods and compositions are provided for nuclease-mediated gene editing of primary cells without the use of viral mediated delivery. Methods of treatments using edited primary cells are also provided.
Claims21
-332. (canceled)
A method of modifying a patient-derived T cell, comprising:
- a. introducing into the cell a homologous recombination (HR) template nucleic acid sequence comprising:
- i. first and second homology arms homologous to first and second endogenous sequences of the cell;
- ii. a T cell receptor (TCR) gene sequence positioned between the first and second homology arms;
- and
- iii. a first 2A-coding sequence positioned upstream of the TCR gene sequence and a second 2A-coding sequence positioned downstream of the TCR gene sequence, wherein the first and second 2A-coding sequences code for the same amino acid sequence but are codon-diverged relative to each other;
- b. recombining the HR template nucleic acid into an endogenous locus of the cell comprising the first and second endogenous sequences homologous to the first and second homology arms of the HR template nucleic acid.
3 more elements
A modified cell comprising an exogenous nucleic acid sequence incorporated into an endogenous locus comprising:
- a. a TCR gene sequence;
- and
- b. a first 2A-coding sequence positioned upstream of the TCR gene sequence and a second 2A-coding sequence positioned downstream of the TCR gene sequence, wherein the first and second 2A-coding sequences code for the same amino acid sequence but are codon-diverged relative to each other.
A method of treating cancer in a human patient comprising:
- a. administering to the human patient a therapeutically effective population of modified cells, wherein the cells comprise an exogenous nucleic acid sequence incorporated into an endogenous locus, the exogenous nucleic acid comprising:
- i. a TCR gene sequence coding for at least a portion of a TCR capable of binding an antigen present on a cancerous cell, and
- ii. a first 2A-coding sequence positioned upstream of the TCR gene sequence and a second 2A-coding sequence positioned downstream of the TCR gene sequence, wherein the first and second 2A-coding sequences are codon-diverged relative to each other, and
- b. thereby treating the cancer in the human patient.
1 more element
Dependencies are inferred from the claim text.