Nat Protoc. 2006;1(6):3049-55.

Retrovirus-mediated single-cell gene knockout technique in adult newborn neurons in vivo.

Source

Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.

Abstract

Single-cell genetic manipulation in an intact brain environment is an informative approach to study molecular mechanism of adult neurogenesis. Here, we describe a protocol for retrovirus-mediated single-cell gene knockout in adult new neurons in vivo. A gene of interest is disrupted in adult floxed mice by a vector based on the Moloney murine leukemia retrovirus, expressing Cre recombinase. High-titer retrovirus is prepared by transfecting plasmids into the HEK293T cells and by concentrating the supernatant containing virus. The retrovirus is stereotaxically injected into the dentate gyrus. Cre recombinase is transduced and expressed in a small fraction of adult new neurons in an intact environment, and the gene knockout is highly efficient within the transduced neurons. Virus preparation takes 7 days, but virus injections take less than 1 h per mouse. By changing the survival time of the mice after the injection, one can analyze the effects on new neurons at different ages.

PMID:: 17406567; [PubMed - indexed for MEDLINE]

MeSH Terms, Substances

MeSH Terms

Substances

LinkOut - more resources

Full Text Sources

Other Literature Sources

COS Scholar Universe

Supplemental Content

Save items

Related citations in PubMed

Efficient CPP-mediated Cre protein delivery to developing and adult CNS tissues. [BMC Biotechnol. 2009]
Efficient CPP-mediated Cre protein delivery to developing and adult CNS tissues.
Gitton Y, Tibaldi L, Dupont E, Levi G, Joliot A. BMC Biotechnol. 2009 Apr 24; 9:40. Epub 2009 Apr 24.
A Cre-lox approach for transient transgene expression in neural precursor cells and long-term tracking of their progeny in vitro and in vivo. [BMC Dev Biol. 2007]
A Cre-lox approach for transient transgene expression in neural precursor cells and long-term tracking of their progeny in vitro and in vivo.
Geoffroy CG, Raineteau O. BMC Dev Biol. 2007 May 15; 7:45. Epub 2007 May 15.
Neuron-specific recombination by Cre recombinase inserted into the murine tau locus. [Biochem Biophys Res Commun. 2008]
Neuron-specific recombination by Cre recombinase inserted into the murine tau locus.
Muramatsu K, Hashimoto Y, Uemura T, Kunii M, Harada R, Sato T, Morikawa A, Harada A. Biochem Biophys Res Commun. 2008 Jun 6; 370(3):419-23. Epub 2008 Mar 31.
Review Cre/loxP-mediated chromosome engineering of the mouse genome. [Handb Exp Pharmacol. 2007]
Review Cre/loxP-mediated chromosome engineering of the mouse genome.
Brault V, Besson V, Magnol L, Duchon A, Hérault Y. Handb Exp Pharmacol. 2007; (178):29-48.
Review Production of a reporter transgenic pig for monitoring Cre recombinase activity. [Biochem Biophys Res Commun. 2009]
Review Production of a reporter transgenic pig for monitoring Cre recombinase activity.
Li L, Pang D, Wang T, Li Z, Chen L, Zhang M, Song N, Nie D, Chen Z, Lai L, et al. Biochem Biophys Res Commun. 2009 May 1; 382(2):232-5. Epub 2009 Mar 5.

See reviews... See all...

Cited by 11 PubMed Central articles

Regulation of adult neurogenesis in the hippocampus by stress, acetylcholine and dopamine. [J Nat Sci Biol Med. 2011]
Regulation of adult neurogenesis in the hippocampus by stress, acetylcholine and dopamine.
Veena J, Rao BS, Srikumar BN. J Nat Sci Biol Med. 2011 Jan; 2(1):26-37.
SoxC transcription factors are required for neuronal differentiation in adult hippocampal neurogenesis. [J Neurosci. 2012]
SoxC transcription factors are required for neuronal differentiation in adult hippocampal neurogenesis.
Mu L, Berti L, Masserdotti G, Covic M, Michaelidis TM, Doberauer K, Merz K, Rehfeld F, Haslinger A, Wegner M, et al. J Neurosci. 2012 Feb 29; 32(9):3067-80.
Impaired spatial and contextual memory formation in galectin-1 deficient mice. [Mol Brain. 2011]
Impaired spatial and contextual memory formation in galectin-1 deficient mice.
Sakaguchi M, Arruda-Carvalho M, Kang NH, Imaizumi Y, Poirier F, Okano H, Frankland PW. Mol Brain. 2011 Sep 1; 4:33. Epub 2011 Sep 1.

See all...

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

Retrovirus-mediated single-cell gene knockout technique in adult newborn neurons...
Retrovirus-mediated single-cell gene knockout technique in adult newborn neurons in vivo.
Nat Protoc. 2006 ;1(6):3049-55.

PubMed
Application of highly sensitive fluorescent dyes (CyDye DIGE Fluor saturation dy...
Application of highly sensitive fluorescent dyes (CyDye DIGE Fluor saturation dyes) to laser microdissection and two-dimensional difference gel electrophoresis (2D-DIGE) for cancer proteomics.
Nat Protoc. 2006 ;1(6):2940-56.

PubMed
An optimized protocol for protein purification in cultured mammalian cells using...
An optimized protocol for protein purification in cultured mammalian cells using a tandem affinity purification approach.
Nat Protoc. 2006 ;1(6):2820-7.

PubMed
The evolution and importance of spinal and chiropractic research.
The evolution and importance of spinal and chiropractic research.
J Manipulative Physiol Ther. 1992 Jan ;15(1):31-5.

PubMed
A protocol for TILLING and Ecotilling in plants and animals.
A protocol for TILLING and Ecotilling in plants and animals.
Nat Protoc. 2006 ;1(5):2465-77.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to: