TY - JOUR
T1 - Mouse Gene-Targeting Strategies for Maximum Ease and Versatility
AU - Papaioannou, Virginia E.
AU - Behringer, Richard R.
N1 - Publisher Copyright:
© 2024 Cold Spring Harbor Laboratory Press.
PY - 2024/1
Y1 - 2024/1
N2 - Well-planned strategies are an essential prerequisite for any mutational analysis involving gene targeting. Consideration of the advantages or disadvantages of different methods will aid in the production of a final product that is both technically feasible and versatile. Strategies for gene-targeting experiments in the mouse are discussed, including the rationale behind some of the common elements of gene-targeting vectors, such as homologous DNA and the use of different site-specific recombinases. We detail positive and negative selection as well as screening strategies for homologous recombination events in embryonic stem (ES) cells. For the planning stages of making different types of alleles, we first consider general strategies and then provide details specific to either homologous recombination in ES cells or making alleles by gene editing with CRISPR-Cas in preimplantation embryos. The types of alleles considered are null or knockout alleles, reporter gene knock-in alleles, point mutations, and conditional null alleles.
AB - Well-planned strategies are an essential prerequisite for any mutational analysis involving gene targeting. Consideration of the advantages or disadvantages of different methods will aid in the production of a final product that is both technically feasible and versatile. Strategies for gene-targeting experiments in the mouse are discussed, including the rationale behind some of the common elements of gene-targeting vectors, such as homologous DNA and the use of different site-specific recombinases. We detail positive and negative selection as well as screening strategies for homologous recombination events in embryonic stem (ES) cells. For the planning stages of making different types of alleles, we first consider general strategies and then provide details specific to either homologous recombination in ES cells or making alleles by gene editing with CRISPR-Cas in preimplantation embryos. The types of alleles considered are null or knockout alleles, reporter gene knock-in alleles, point mutations, and conditional null alleles.
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U2 - 10.1101/pdb.over107957
DO - 10.1101/pdb.over107957
M3 - Review article
C2 - 37932102
AN - SCOPUS:85181760672
SN - 1940-3402
VL - 2024
JO - Cold Spring Harbor protocols
JF - Cold Spring Harbor protocols
IS - 1
ER -