Disclosed are compositions and methods for
in vivo genetic engineering of
lymphocyte precursors. A lipid
nanoparticle (LNP), surface-functionalized for
lymphocyte precursor targeting, encapsulates
a DNA payload comprising a therapeutic
gene flanked by engineered Recombination
Signal Sequences (
RSS). The
RSS spacer incorporates at least one non-natural
nucleotide, reducing integration risks. Because RAG1 and RAG2 are expressed only after commitment to the lymphoid lineage, integration is restricted to immune precursors and cannot occur in stem cells or
reproductive cells. Upon systemic delivery, the endogenous RAG1 / RAG2 complex integrates the
payload into the precursor
genome. Modified precursors undergo clonal expansion
in vivo, amplifying the
therapeutic effect from relatively low doses. In certain embodiments, the
DNA payload encodes receptors or antibodies that specifically recognize aggregated
amyloid proteins, including
transthyretin (TTR) fibrils and immunoglobulin light- chain aggregates, enabling
therapeutic treatment of
amyloidosis. However, the platform is not limited to
amyloidosis and can be applied to
oncology, infectious diseases, autoimmune disorders, and other
protein misfolding conditions.