Microinjection of Xenopus oocytes

TRISTÁN AGÜERO; KAREN NEWMAN; MARY LOU KING

Early Development of Xenopus laevis: A Laboratory Manual

Cold Spring Harbor

Año: 2018; p. 20 - 28

Microinjection of Xenopus oocytes has proven to be avaluable tool in a broad range of studies that require expression of DNA or RNAinto functional protein. These studies are diverse ranging from expressioncloning to receptor-ligand interaction to nuclear programming.  See Table 1. Oocytes offer a number of advantages for these studies including theirlarge size (~1.2 mm in diameter), their capacity for translation, and enormousnucleus (0.3-0.4mm). They are cost effective, easily manipulated, and largenumbers can be injected in a short time period. Oocytes have a large maternalstockpile of all the essential components for transcription and translation.  Consequently, the investigator needs only tointroduce by microinjection, their specific DNA or RNA for synthesis.  Oocytes will translate virtually any exogenousRNA regardless of source, and the translated proteins are folded, modified, andtransported to correct cellular locations. Here we present procedures for the efficient microinjection of oocytesand their subsequent care.