TY - JOUR

T1 - Concatenation of transgenic DNA: Random or orchestrated?

AU - Smirnov, Alexander

AU - Battulin, Nariman

N1 - Funding Information: Funding: This work was supported by Russian Foundation for Basic Research, grant number 20-34-70087. N.B. supported from the Ministry of Education and Science of Russian Federation, grant #2019-0546 (FSUS-2020-0040). Data analysis performed on computational nodes of the Institute of Cytology and Genetics (budget project no. 0259-2021-0016). Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2021/12

Y1 - 2021/12

N2 - Generation of transgenic organisms by pronuclear microinjection has become a routine procedure. However, while the process of DNA integration in the genome is well understood, we still do not know much about the recombination between transgene molecules that happens in the first moments after DNA injection. Most of the time, injected molecules are joined together in head-to-tail tandem repeats—the so-called concatemers. In this review, we focused on the possible concatenation mechanisms and how they could be studied with genetic reporters tracking individual copies in concatemers. We also discuss various features of concatemers, including palindromic junctions and repeat-induced gene silencing (RIGS). Finally, we speculate how cooperation of DNA repair pathways creates a multicopy concatenated insert.

AB - Generation of transgenic organisms by pronuclear microinjection has become a routine procedure. However, while the process of DNA integration in the genome is well understood, we still do not know much about the recombination between transgene molecules that happens in the first moments after DNA injection. Most of the time, injected molecules are joined together in head-to-tail tandem repeats—the so-called concatemers. In this review, we focused on the possible concatenation mechanisms and how they could be studied with genetic reporters tracking individual copies in concatemers. We also discuss various features of concatemers, including palindromic junctions and repeat-induced gene silencing (RIGS). Finally, we speculate how cooperation of DNA repair pathways creates a multicopy concatenated insert.

KW - Concatemers

KW - Double-strand breaks (DSBs)

KW - Homologous recombination (HR)

KW - Non-homologous end-joining (NHEJ)

KW - Palindromes

KW - Pronuclear microinjection

KW - Repeat-induced gene silencing (RIGS)

KW - Transgenic animals

UR - http://www.scopus.com/inward/record.url?scp=85121420198&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/85eecbca-fc65-3522-8701-d51e55dd8c40/

U2 - 10.3390/genes12121969

DO - 10.3390/genes12121969

M3 - Review article

C2 - 34946918

AN - SCOPUS:85121420198

VL - 12

JO - Genes

JF - Genes

SN - 2073-4425

IS - 12

M1 - 1969

ER -