Abstract：
:Targeted insertion of large pieces of DNA is an important goal of genetic engineering. However, this goal has been elusive since classical methods for homology-directed repair are inefficient and often not feasible in many systems. Recent advances are described here that enable site-specific genomic insertion of relatively large DNA with much improved efficiency. Using the preferred repair pathway in the cell of nonhomologous end-joining, DNA of up to several kb could be introduced with remarkably good precision by the methods of HITI and ObLiGaRe with an efficiency up to 30-40%. Recent advances utilizing homology-directed repair (methods of PITCh; short homology arms including ssODN; 2H2OP) have significantly increased the efficiency for DNA insertion, often to 40-50% or even more depending on the method and length of DNA. The remaining challenges of integration precision and off-target site insertions are summarized. Overall, current advances provide major steps forward for site-specific insertion of large DNA into genomes from a broad range of cells and organisms.
journal_name
Chromosomajournal_title
Chromosomaauthors
Yamamoto Y,Gerbi SAdoi
10.1007/s00412-018-0677-6subject
Has Abstractpub_date
2018-12-01 00:00:00pages
405-420issue
4eissn
0009-5915issn
1432-0886pii
10.1007/s00412-018-0677-6journal_volume
127pub_type
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