Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes

PloS One
Youmei WuJames M Pickel

Abstract

Design and engineering of complex knockin mice has revolutionized the in vivo manipulation of genetically defined cells. Recently development of the bacterial clustered regularly interspersed short palindromic repeats (CRISPR) associated protein 9 (Cas9) system for single site cleavage of mammalian genomes has opened the way for rapid generation of knockin mice by targeting homology directed repair to selected cleavage sites. We used this approach to generate new lines of mice that will be useful for a variety of aspects of neuroscience research. These lines have been bred to homozygosity and details of the expression and function of the transgenes are reported. Two lines target the Rosa26-locus and have been engineered to allow Cre-dependent expression of the avian tva receptor, and Cre-dependent expression of a cell surface targeted spaghetti-monster carrying many copies of the "ollas-tag". Another line expresses red fluorescent protein and tva in Tac1-positive neurons; the fourth line targets FlpO expression to Plekhg1 expressing neurons, providing a powerful approach to modify gene expression in thalamic excitatory neurons.

References

Aug 18, 2001·Cell·G NelsonC S Zuker
Dec 21, 2006·Nature Methods·Ian R WickershamEdward M Callaway
Sep 4, 2007·Cellular and Molecular Life Sciences : CMLS·J WuS Chandrasegaran
Dec 9, 2010·The EMBO Journal·Santosh K MishraMark A Hoon
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jan 5, 2013·Science·Le CongFeng Zhang
Jan 5, 2013·Science·Prashant MaliGeorge M Church
Feb 7, 2013·ELife·Martin JinekJennifer Doudna
Jul 28, 2013·Nature Protocols·Fumitaka Osakada, Edward M Callaway
May 28, 2014·Nucleic Acids Research·Tessa G MontagueEivind Valen
Jul 22, 2014·The FEBS Journal·Yuanwu MaLianfeng Zhang
Jul 25, 2014·Nature Protocols·Hui YangRudolf Jaenisch
Jul 30, 2014·Frontiers in Neural Circuits·Julie A HarrisHongkui Zeng
Nov 20, 2014·Methods in Molecular Biology·Takehito Kaneko, Tomoji Mashimo
Apr 29, 2015·Nature Methods·Sarada ViswanathanLoren L Looger
Nov 19, 2015·Nature·Yueqing PengCharles S Zuker
Jan 9, 2016·Endocrine Journal·Takuro Horii, Izuho Hatada
May 18, 2016·Nucleic Acids Research·Kornel LabunEivind Valen
Sep 13, 2016·Cell·Chan Lek TanZachary A Knight
Jun 24, 2017·Methods in Molecular Biology·Tetsushi Sakuma, Takashi Yamamoto

❮ Previous
Next ❯

Citations

Nov 14, 2020·Neuron·Lars J von BuchholtzNicholas J P Ryba

❮ Previous
Next ❯

Methods Mentioned

BETA
gene
targeted mutations
gene-knockout
transgenic
PCR
restriction digest
Assay
Fluorescence

Software Mentioned

Adobe Photoshop

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.