PMID- 9859218
OWN - NLM
STAT- MEDLINE
DCOM- 19990119
LR  - 20191102
IS  - 0962-8819 (Print)
IS  - 0962-8819 (Linking)
VI  - 7
IP  - 4
DP  - 1998 Jul
TI  - Nuclear localization signals enhance germline transmission of a transgene in 
      zebrafish.
PG  - 303-9
AB  - We report that cytoplasmic injection into zebrafish eggs of 10(4) copies of 
      plasmid DNA complexed to nuclear localization signal (NLS) peptides, as compared 
      to 10(6) copies of naked DNA, increased nuclear uptake of transgene DNA early 
      during embryo development and enhanced transgene integration frequency into the 
      germline of founders. Monitoring the dynamics of nuclear uptake of DNA-NLS 
      complexes by fluorescence in situ hybridization (FISH) of interphase nuclei 
      indicates that NLS enhances both the proportion of nuclei importing DNA during 
      early embryo development, and the amount of DNA imported by individual nuclei. 
      The use of NLS increases the proportion of germline transgenic founders from 14 
      to 43% (P < 0.01) as assessed by polymerase chain reaction analysis of F1s. From 
      germline transgenic DNA-NLS-injected founders, 47% transgenic F1s are obtained in 
      wild-type crosses, as opposed to 6% from naked DNA-injected founders (P < 0.01). 
      In both cases, the transgene is transmitted to the F2 generation. In addition, 
      high-resolution FISH analysis of transgenic F1s reveals that the use of NLS 
      increases the number of distinct transgene integration sites along chromatin 
      fibres.
FAU - Collas, P
AU  - Collas P
AD  - Department of Biochemistry, Norwegian College of Veterinary Medicine, Oslo, 
      Norway.
FAU - Alestrom, P
AU  - Alestrom P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Transgenic Res
JT  - Transgenic research
JID - 9209120
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Recombinant)
RN  - 0 (Nuclear Localization Signals)
SB  - IM
MH  - Animals
MH  - Animals, Genetically Modified
MH  - Base Sequence
MH  - Binding Sites
MH  - Cell Nucleus/metabolism
MH  - DNA Primers/genetics
MH  - DNA, Recombinant/genetics/metabolism
MH  - Female
MH  - In Situ Hybridization, Fluorescence
MH  - Nuclear Localization Signals/*genetics
MH  - Plasmids/genetics/metabolism
MH  - Zebrafish/*genetics
EDAT- 1998/12/22 00:00
MHDA- 1998/12/22 00:01
CRDT- 1998/12/22 00:00
PHST- 1998/12/22 00:00 [pubmed]
PHST- 1998/12/22 00:01 [medline]
PHST- 1998/12/22 00:00 [entrez]
AID - 10.1023/a:1008826230315 [doi]
PST - ppublish
SO  - Transgenic Res. 1998 Jul;7(4):303-9. doi: 10.1023/a:1008826230315.