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You see, cam videos can last for hours sometimes. In recent years, a strategy for transgene integration has been developed that uses cleavage with site-specific nucleases for targeted insertion into a chosen genomic locus (see, e.g., U.S. Nos. 9,877,988; 9,567,573; 9,394,545; 9,222,105; 9,150,847; 7,888,121; 7,972,854; 7,914,796; 7,951,925; 8,895,264; 8,771,985; 8,110,379; 8,409,861; 8,586,526; U.S. Pat. Nos. 7,166,745; 7,173,154; 7,323,594; 7,470.817; 7,479,573; 7,601,872; 7,915,450; 8,158,827; 8,785,200; 9,358.300, International Patent Publication No. WO 2016/011203, and U.S. Patent Publication No. 2017/0107539, all of which are hereby expressly incorporated by reference in their entirety as though fully set forth herein. Lipid nanoparticles (LNP) are one of the most promising types of nanoparticles due to high encapsulating efficiency of nucleic acids, high stability and compatibility with biologic environments (Lee et al. The present disclosure provides methods and compositions to increase the efficiency of gene therapy in pulmonary tissue through the use of LNP reagents comprising gene therapy reagents

In some embodiments, the transgene encodes a protein for treatment of a patient in need thereof and is integrated using the methods and compositions of the invention into a safe harbor locus. In some embodiments, the composition is administered to a patient and then re-administered 7, 14, 21, 28, 30, 40, 50, 75, 100, 200 or more days after the first administration. The term also applies to amino acid polymers in which one or more amino acids are chemical analogues or modified derivatives of a corresponding naturally-occurring amino acids. In some embodiments, the nucleic acid encodes an engineered nuclease wherein the nuclease comprises a DNA binding domain and a cleavage domain. In some embodiments, the DNA cleavage domain is FokI. In other embodiments, the DNA-binding domain is a TALE DNA-binding domain. These are single chain fusion proteins linking a TALE DNA binding domain to a TevI nuclease domain. Especially preferred are compositions comprising an LNP comprising mRNAs encoding a nuclease, and an AAV comprising a DNA donor. In another aspect, fusion polypeptides comprising a DNA binding domain and an engineered cleavage half-domain as described herein are provided

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