Transgenic mice
The invention relates to nucleic acid constructs for expression in mice for the production of heavy chain only antibodies and V H domains, transgenic mice, related methods and uses.
1. A transgenic mouse comprising in its genome a vector comprising:
a) at least 10 functional human heavy chain V genes wherein at least 10 functional human heavy chain V genes are in their natural configuration;
b) at least one human heavy chain D gene and at least one human heavy chain J gene;
c) a murine μ enhancer;
d) a murine switch μ element;
e) a murine C gene which lacks the CH1 exon, wherein the C gene consists of a C gene selected from Cγ1, Cγ2b and/or Cγ2a; and
f) a murine 3′ enhancer region,
wherein Cμ is absent in the mouse, or Cμ is partially absent to the extent that it is non-functional, and
wherein the transgenic mouse is capable of expressing a chimeric immunoglobulin heavy chain encoded by the vector in response to antigen challenge.
2. A transgenic mouse according to claim 1 wherein said mouse comprises a disruption in one or more endogenous immunoglobulin loci, wherein the disruption renders the one or more endogenous immunoglobulin loci non-functional.
3. A transgenic mouse according to claim 2 wherein said mouse comprises a non-functional endogenous lambda light chain locus and/or a non-functional endogenous kappa light chain locus and/or a non-functional endogenous heavy chain locus.
4. A method for making an in vitro display library comprising the step of using a transgenic mouse according to claim 1 to produce an in vitro display library.
5. A method according to claim 4 wherein said library is a naïve library.
6. A method for making a library comprising the step of immunisation of a transgenic mouse according to claim 1 .
7. A method according to claim 4 , comprising the steps of;
a) isolating a cell or tissue expressing a heavy chain antibody (HCAb),
b) cloning the sequence encoding the V H domain from mRNA derived from the isolated cell or tissue,
c) constructing a library from cloned transcripts, and
d) isolating the V H domain.
8. A method according to claim 7 , wherein the method comprises using next generation sequencing to construct the library of cloned transcripts.
9. A method according to claim 6 , comprising the steps of;
a) isolating a cell or tissue expressing a heavy chain antibody (HCAb),
b) cloning the sequence encoding the V H domain from mRNA derived from the isolated cell or tissue,
c) constructing a library from cloned transcripts, and
d) isolating the V H domain.
10. A method according to claim 9 , wherein the method comprises using next generation sequencing to construct the library of cloned transcripts.
11. The transgenic mouse according to claim 2 , wherein the disruption is a partial deletion, a complete deletion, or an insertion.